KR20170012139A - biomarker comprising fbp gene in common associating aging, cancer and obesity and diagnosis kit using thereof - Google Patents

Abstract

The present invention relates to a fructose-1,6-bisphosphatase (fbp, CG31692) gene mutually related to aging, obesity, and cancer. More specifically, the present invention relates to a diagnosing kit and the like using the gene. The gene and the diagnosing kit according to the present invention discover a fbp gene mutually related to the cause and occurrence of aging, obesity, and cancer, and provided is a biomarker for making it possible to analyze the progress of aging, obesity, and cancer by using the same. Consequently, the progress of aging, the occurrence of cancer, and the occurrence of obesity in a human, a mammal excluding a human, or an insect can be comprehensively analyzed or diagnosed.

Description

Biomarkers comprising the fbp gene commonly associated with aging, obesity and cancer, and uses thereof, and their use,

The present invention relates to fbp (Fructose-1,6-bisphosphatase, CG31692) gene commonly associated with aging, obesity and cancer, and more specifically relates to a diagnostic kit using the gene.

 Studies on genes regulating senescence began in the early 1990s and are now actively under way. The genes regulating senescence so far have been classified into functional groups: catalase, superoxide dismutase, insulin / IGF-1 signaling pathway (including insulin, InR, PI3K, Akt and Foxo) (sirtuins), cancer suppression systems (p53, etc.), material delivery systems (sodium dicarboxylate cotransporter, etc.), and telomere control systems. These genes are closely involved in the regulation of senescence.

On the other hand, when aging progresses, cancer development also increases. Interestingly, cancer suppressor genes such as p53 regulate senescence at the same time. (Ras, Rac, Rap, Rala, Rhoa, etc.), Akt related genes (Akt / PKB, PKC, PKA, RAF, etc.) with serine / threonine kinase activity, , hedgehog-related genes, and protooncogene (c-Myc), and p53 and NFkB as cancer-suppressing genes. In particular, HGF and its receptor HGFR (c-met) are mainly associated with liver cancer. In addition, PTEN, a cancer-suppressing gene, inhibits the activity of PI3K. Overexpression of PTEN reduces the activity of the insulin / IGF-1 signal transduction system and increases its lifespan. In addition, all of these genes have been found in model organisms such as yeast, nematodes, Drosophila and mice, and research on human genes having functions to control cancer and aging in general has not been well studied yet.

Also, as aging progresses, obesity tends to occur. The causes of obesity related to aging include lack of exercise, growth hormone (GH) and hypothyroid hormone secretion. When the secretion of GH decreases, the metabolic effects of GH degrading carbohydrates, fats, Production is reduced and fat accumulation is induced. GH increases the secretion of IGF-1 in the liver, and thus, the decrease in GH secretion following aging will alter the activity of the insulin / IGF-1 signaling pathway and will be involved in the regulation of life span. For example, in the case of FIRKO mice in which insulin receptor is removed from adipocytes, fat accumulation may not occur even when overeating, and the body fat content of the fruit fly, an animal model of senescence, increases as aging proceeds . Therefore, despite the fact that obesity and aging are closely related, research on human genes that have the function of controlling aging and obesity in general is scarce.

In order to develop a biomarker, a diagnostic kit, a screening method, and the like that can discover genes that commonly control aging, cancer, and obesity, and can commonly diagnose aging, cancer, and obesity by confirming the expression of these genes There is a problem that the research is insufficient now.

On the other hand, Korean Patent Laid-Open Publication No. 10-2012-0021401 (Patent Document 1) has been disclosed as a prior art document related to the present invention, and Patent Document 1 discloses that the Atg5 gene is over- There is no disclosure or suggestion about a biomarker, a diagnostic kit and a screening method using a gene that commonly controls aging, cancer, and obesity, and the like, .

Patent Document 1. Korean Patent Laid-Open No. 10-2012-0021401

DISCLOSURE OF THE INVENTION The object of the present invention is to solve the above-mentioned problems, and it is an object of the present invention to provide a method for detecting a gene commonly involved in aging, cancer and obesity, And to provide a biomarker capable of quickly and accurately diagnosing cancer.

The present invention also provides a kit using the same and a method of discriminating or diagnosing the same.

In order to accomplish the above object, the present invention provides a nucleic acid comprising a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, a complementary base sequence thereof, There is provided a biomarker for discriminating the progress of aging comprising a single nucleotide sequence.

As the aging of the diagnostic subject is promoted, the expression amount of the biomarker is reduced.

In order to accomplish the above other objects, the present invention provides a DNA sequence comprising a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, a complementary base sequence thereof, A biomarker for obesity discrimination comprising any one of the base sequences is provided.

The amount of expression of the biomarker is decreased as the fat accumulation of the diagnostic subject is increased and the obesity is increased.

According to another aspect of the present invention, there is provided a recombinant DNA comprising a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, a complementary base sequence thereof, A cancer diagnostic biomarker comprising any one of the base sequences is provided.

And the expression amount of the biomarker is decreased as the occurrence of cancer cells or cancer tissues of the diagnostic individual is increased.

According to another aspect of the present invention, there is provided a biomarker for determining aging progress. And a hybridization solution.

The biomarker may be present in a dispersed form in solution. And is present in the form of a microarray immobilized on a substrate.

According to another aspect of the present invention, there is provided a biomarker for obesity discrimination. And a hybridization solution.

The biomarker may be present in a dispersed form in solution. And is present in the form of a microarray immobilized on a substrate.

According to another aspect of the present invention, there is provided a biomarker for cancer diagnosis. And a hybridization solution.

The biomarker may be present in a dispersed form in solution. And is present in the form of a microarray immobilized on a substrate.

According to another aspect of the present invention, there is provided a biomarker comprising: And a hybridization solution; and a kit for complex diagnosis that simultaneously detects an aging process discrimination, an obesity discrimination, and a cancer diagnosis.

According to another aspect of the present invention, there is provided an aging process discriminating method comprising the steps of:

I) isolating and extracting RNA from a diagnostic individual;

II) contacting the isolated RNA or cDNA synthesized therefrom with the aging-progression-determining kit to hybridize the RNA or cDNA with the biomarker; And

III) detecting the degree of hybridization between the biomarker and the RNA or cDNA.

According to another aspect of the present invention, there is provided an obesity discrimination method comprising the steps of:

I) isolating and extracting RNA from a diagnostic individual;

II) contacting the isolated RNA or the synthesized cDNA with the obesity-determining kit to hybridize the RNA or cDNA with the biomarker; And

III) detecting the degree of hybridization between the biomarker and the RNA or cDNA.

According to another aspect of the present invention, there is provided a cancer diagnosis method comprising the steps of:

I) isolating and extracting RNA from a diagnostic individual;

II) contacting the isolated RNA or the synthesized cDNA with the cancer diagnostic kit to hybridize the RNA or cDNA with the biomarker; And

III) detecting the degree of hybridization between the biomarker and the RNA or cDNA.

According to another aspect of the present invention, there is provided a method for simultaneously detecting an aging process discrimination, an obesity increase discrimination, and a cancer diagnosis.

I) isolating and extracting RNA from a diagnostic individual;

II) contacting the isolated RNA or cDNA synthesized therefrom with the complex diagnostic kit to hybridize the RNA or cDNA with the biomarker; And

III) detecting the degree of hybridization between the biomarker and the RNA or cDNA.

The gene and the diagnostic kit according to the present invention find fbp (Fructose-1,6-bisphosphatase, CG31692) gene commonly involved in aging, obesity and cancer, To be analyzed. Thus, it is possible to comprehensively analyze or diagnose whether progress of human aging, cancer occurrence, and obesity has occurred.

Figure 1 is a graph showing Actin-GS-Gal4 / + W1118 lifetime curves for RU486 treatment.
Fig. 2 shows the results of the decrease in the amount of fbp mRNA when Actin-GS-Gal4 and UAS-fbp RNAi were used to inhibit the expression of fbp.
FIG. 3 is a graph showing a result of confirming that the lifespan is shortened when the expression of fbp is decreased.
FIG. 4 shows the change in the triglyceride content of wild-type fruit fly fed with RU486.
FIG. 5 shows the results of the addition of Actin-GS-Gal4 / +; UAS-fbp RNAi / + Drosophila.
Fig. 6 is a confocal microscopic photograph of the fat body tissue of Actin-GS-Gal4 / UAS-nls. GFP fruit flies fed with RU486.
FIG. 7 is a photograph of a confocal microscope taken after Nil red staining of the fat tissue of Actin-GS-Gal4 / UAS-fbp RNAi fruit fly fed with RU486.
8 shows the results of comparison of the blade lengths of the cancer growth model flies (UAS-Ras85D; c765-Gal4).
Fig. 9 shows the results of comparison of the blade lengths of cancer growth model flies (UAS-Ras85D / +; c765-Gal4 / UAS-fbp RNAi) inhibiting fbp gene expression.
Fig. 10 shows the result of comparison of the wing area of the cancer-proliferation model flies (UAS-Ras85D / +; c765-Gal4 / UAS-fbp RNAi) inhibiting the expression of fbp gene.

Accordingly, the present inventors have intensively studied to find a gene commonly involved in aging, obesity and cancer. As a result, it has been found that fbp (Fructose-1,6-bisphosphatase, CG31692), which is commonly involved in aging, obesity, Gene and discovered the present invention.

It is an object of the present invention to provide a biomarker capable of promptly and simply discriminating the progress of aging of a mammal or an insect other than human or human.

One aspect of the present invention is a nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, a complementary base sequence thereof, And a biomarker for discriminating the progress of aging comprising at least one base sequence.

A gene consisting of a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, a complementary base sequence thereof and an mRNA thereof The sequence decreases as the aging, which is a phenotype of the life span of the diagnostic individual, is promoted.

Thus, using this characteristic, the nucleotide sequence is a biomarker capable of discriminating the progress of aging by comparing the standard amount of the nucleotide sequence in the same species as that of the diagnostic individual and the expression amount of the nucleotide sequence in the diagnostic individual Can be utilized.

In this specification, a biomarker for determining the progress of aging means a biomarker capable of qualitatively discriminating the life span, that is, the progress of aging.

As described above, the mammalian animals or insects except humans or humans according to the present invention have genetic differences with each other, and the average degree of progression of aging is completely different. However, any one selected from the group consisting of a base sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, a complementary base sequence thereof, By providing the above base sequence as a biomarker for determining the progress of aging, it is possible to quickly, accurately, and simply discriminate the progress of aging of the diagnostic object with respect to each species.

Any one of the nucleotide sequences selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4 is commonly used in the UAS-GAL4 system The nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 is preferably fbp (Fructose-1,6- bisphosphatase, CG31692) gene.

In the present invention, GAL / UAS expression system was used for evaluating the function of candidate genes using Drosophila. Specifically, a Gene Switch (GS) GAL / UAS expression system was used. GAL4 was originally a yeast protein. GAL4, which is expressed by introducing it into Drosophila, binds to a DNA sequence called Upstream Activating Sequencers (UAS) when a drug called RU486 (mifepristone) is present and activates the transcription of a specific gene after UAS , And transcription of a specific gene is inactivated if RU486 is not present. Using this, the fbp gene expression was regulated by regulating the activity of fbp RNAi, and its function was confirmed.

Any one or more of the base sequences selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, their complementary base sequences and mRNAs thereof, Reduced expression in the liver promotes the aging of fruit flies.

Specifically, after the expression of the gene of the UAS-GAL4 gene has been exposed to RU486, the effect of decreasing the expression of the specific gene by more than 2-fold was confirmed through repeated experiments. As a result, Any one or more of the base sequences selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, their complementary base sequences and their mRNAs When the expression of the contained gene is reduced more than twice, it is confirmed that the aging progresses further. The above-mentioned gene can be used as a biomarker for discriminating the aging of a mammal or an insect other than human or human.

Furthermore, any one or more base sequences selected from the group consisting of any one of base sequences selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, their complementary base sequences, Has a homologue relationship with human ADHFE1 (alcohol dehydrogenase, iron containing, 1, NP_653251.2 467 aa) gene. Therefore, it is also judged whether or not the progress of aging of human beings as well as insects is made using the biomarker according to the present invention can do.

Any one or more of the base sequences selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, and complementary base sequences thereof, And these cDNAs are prepared from complementary DNA using mRNA produced through transcription from genomic DNA.

That is, when the biomarker is used, the progress of aging of mammals and insects other than humans and humans can be discriminated, and thus it is expected to be useful for discriminating and responding to the progress of aging of diagnostic objects.

It is an object of the present invention to provide a biomarker capable of promptly and simply discriminating obesity of a mammal or an insect other than human or human.

One aspect of the present invention is a nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, a complementary base sequence thereof, To a biomarker for obesity discrimination comprising one or more base sequences.

A gene consisting of a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, a complementary base sequence thereof and an mRNA thereof The sequence decreases with the increase in the obesity of the diagnostic individual (specifically, the increase in the triglyceride content and the increase in the size and density of the fat sphere in the fat body tissue).

Therefore, using the above characteristics, the nucleotide sequence can be used as a biomarker capable of discriminating the increase in obesity by comparing the standard amount of the nucleotide sequence of the same species as that of the diagnostic individual with the expression amount of the nucleotide sequence measured in the diagnostic individual .

In this specification, biomarker for discrimination of obesity means a biomarker capable of qualitatively discriminating the increase in obesity (specifically, increase in triglyceride content and increase in size and density of fat globule in fat body tissue).

As described above, since mammals or insects other than humans or humans according to the present invention have genetic differences with each other, there will be a difference in obesity. However, any one selected from the group consisting of a base sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, a complementary base sequence thereof, By providing the above base sequence as a biomarker for discrimination of obesity, it is possible to quickly, accurately and simply determine the obesity of a diagnostic object for each species.

Any one of the nucleotide sequences selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 is usually extracted from a Drosophila mutant that controls the expression of a specific gene by the UAS-GAL4 system Preferably, the nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4 is fbp (Fructose-1,6-bisphosphatase, CG31692) gene of Drosophila.

In the present invention, GAL / UAS expression system was used for evaluating the function of candidate genes using Drosophila. Specifically, a Gene Switch (GS) GAL / UAS expression system was used. GAL4 was originally a yeast protein. GAL4, which is expressed by introducing it into Drosophila, binds to a DNA sequence called Upstream Activating Sequencers (UAS) when a drug called RU486 (mifepristone) is present and activates the transcription of a specific gene after UAS , And transcription of a specific gene is inactivated if RU486 is not present. Using this, the fbp gene expression was regulated by regulating the activity of fbp RNAi, and its function was confirmed.

Any one or more of the base sequences selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, their complementary base sequences and mRNAs thereof, The expression level decreases specifically as the triglyceride content of Drosophila and the size and density of lipids in fat tissue are greatly increased.

Specifically, after the expression of the gene of the UAS-GAL4 gene has been exposed to RU486, the effect of decreasing the expression of the specific gene by more than 2-fold was confirmed through repeated experiments. As a result, Any one or more of the base sequences selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, their complementary base sequences and their mRNAs It was confirmed that obesity increases when the expression of the gene containing the gene is decreased by a factor of two or more. As a result, the above-mentioned gene can be used as a biomarker for discriminating the obesity of a mammal or insect other than human or human.

Furthermore, any one or more base sequences selected from the group consisting of any one of base sequences selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, their complementary base sequences, Has a homologue relationship with human ADHFE1 (alcohol dehydrogenase, iron containing, 1, NP_653251.2 467 aa) gene, it is possible to discriminate not only insects but also human beings by using the biomarker according to the present invention .

Any one or more of the base sequences selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, and complementary base sequences thereof, And these cDNAs are prepared from complementary DNA using mRNA produced through transcription from genomic DNA.

That is, when the biomarker is used, it is possible to discriminate the triglyceride contents of mammals and insects other than humans and humans, and the size and density of lipids in fat body tissues. .

It is an object of the present invention to provide a biomarker capable of rapidly and simply diagnosing cancer of a mammal or an insect other than human or human.

One aspect of the present invention is a nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, a complementary base sequence thereof, To a cancer diagnostic biomarker comprising one or more base sequences.

A gene consisting of a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, a complementary base sequence thereof and an mRNA thereof The sequence decreases as the cancer occurrence or proliferation (specifically, the phenotype of the cancer growth model Drosophila) increases in the diagnostic individual.

Therefore, by using such a characteristic, the nucleotide sequence can be determined by comparing the standard onset amount of the base sequence in the same species as the diagnostic individual and the expression amount of the determined base sequence in the diagnostic individual to determine a biomarker .

In the present specification, biomarker for cancer diagnosis means a biomarker capable of qualitatively discriminating whether cancer cells and tissues proliferate or grow.

As described above, since the mammalian animal or insect except the human or human according to the present invention has a genetic difference with each other, the occurrence of cancer is completely different. However, any one selected from the group consisting of a base sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, a complementary base sequence thereof, By providing the above base sequence as a biomarker for cancer diagnosis, it is possible to promptly, accurately, and simply determine the occurrence or growth of the cancer in each individual species.

Any one of the nucleotide sequences selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 is usually extracted from a Drosophila mutant that controls the expression of a specific gene by the UAS-GAL4 system Preferably, the nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4 is fbp (Fructose-1,6-bisphosphatase, CG31692) gene of Drosophila.

In the present invention, GAL / UAS expression system was used for evaluating the function of candidate genes using Drosophila. Specifically, a Gene Switch (GS) GAL / UAS expression system was used. GAL4 was originally a yeast protein. GAL4, which is expressed by introducing it into Drosophila, binds to a DNA sequence called Upstream Activating Sequencers (UAS) when a drug called RU486 (mifepristone) is present and activates the transcription of a specific gene after UAS , And transcription of a specific gene is inactivated if RU486 is not present. Using this, the fbp gene expression was regulated by regulating the activity of fbp RNAi, and its function was confirmed.

Any one or more of the base sequences selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, their complementary base sequences and mRNAs thereof, The expression level decreases specifically as the triglyceride content of Drosophila and the size and density of lipids in fat tissue are greatly increased.

Specifically, the effect of the expression of a specific gene on the expression of RU486 after the expression of a specific gene was controlled by the UAS-GAL4 system was more than twice as much as that of the wild-type mutant (SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4), their complementary base sequences and their complementary sequences When the expression of the gene containing any one or more of the nucleotide sequences selected from the group consisting of these mRNAs is decreased more than two times, it has been found that the cancer is increased. The above-mentioned gene can be used for a mammal or an insect And can be used as a biomarker for diagnosing cancer occurrence.

Furthermore, any one or more base sequences selected from the group consisting of any one of base sequences selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, their complementary base sequences, Has a homologue relationship with a human ADHFE1 (alcohol dehydrogenase, iron containing, 1, NP_653251.2 467 aa) gene. Therefore, it is possible to use the biomarker according to the present invention to detect not only insects but also human cancer Can be diagnosed.

Any one or more of the base sequences selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, and complementary base sequences thereof, And these cDNAs are prepared from complementary DNA using mRNA produced through transcription from genomic DNA.

That is, when the above-mentioned biomarker is used, it is possible to discriminate whether or not cancer and growth of mammals and insects other than humans and humans are caused and proliferation.

In addition, any one selected from the group consisting of a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, a complementary base sequence thereof and mRNA thereof The biomarker including one or more base sequences can detect aging discrimination, obesity discrimination, and cancer diagnosis as described above, but can also detect aging progress, obesity increase, and cancer occurrence.

As described above, any one selected from the group consisting of the nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, the complementary base sequence thereof and the mRNA thereof As the expression of the above base sequences is decreased, the aging of the fruit fly mutant which reduces the expression of a specific gene using the UAS-GAl4 system is further progressed, the fat pore density and size in the triglyceride and fat body tissue are increased, It is possible to simultaneously detect progress of aging, increase in obesity and cancer occurrence by comparing the standardized amount of biomarker of the same species as the diagnostic individual and the measured biomarker expression amount in the diagnostic individual .

In the present invention, the biomarker may be any one selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, mRNA of the complementary base sequence, The protein may be any one or more of the amino acid sequences selected from the group consisting of SEQ ID NOS: 5 to 8.

The protein represented by any one of the amino acid sequences selected from the group consisting of SEQ ID NOS: 5 to 8 is also useful as an agent for preventing or treating senescence, obesity and / or inflammation by increasing or decreasing the amount of protein compared to a normal control (standard expression amount of the same species as the diagnostic individual) Cancer can be distinguished and diagnosed.

The kit using the protein may include a kit for ELISA (Enzyme linked immunosorbent assay), and the antibody specifically binding to the protein may be administered to a biological sample selected from a cell, tissue, urine, blood, serum, Through the step of contacting the sample, the antigen-antibody complex can be quantitatively detected.

The detection result may be compared with a standard protein expression amount in a diagnostic subject to discriminate and diagnose any one or more selected from aging, obesity and cancer of the diagnostic subject.

Analysis methods for measuring protein expression include Western blotting, enzyme linked immunosorbent assay (ELISA), radioimmunoassay (RIA), radioimmunodiffusion, Ouchterlony immunodiffusion, rocket immunoelectrophoresis, , Immunoprecipitation, Complement fixation assay, FACS, Protein chip, etc., and are not limited to the described methods.

Through the above analysis method, it is possible to compare the amount of the standard antigen-antibody complex formed in the same species as the diagnostic individual with the amount of the antigen-antibody complex formed in the diagnostic individual, and the significant expression level of the protein expressed by the amino acid sequence 2 , It is possible to discriminate or diagnose any one or more of aging progression, obesity increase, and cancer development.

Another aspect of the present invention relates to a kit for determining the progress of aging comprising the biomarker and the hybridization solution.

The biomarker is as described above, and may be stored in a solution, or the biomarker may be immobilized on the substrate at a high density. In other words, the biomarkers may be in a microarray form immobilized in a predetermined region. Such microarrays are well known in the art.

The kit may be dispersed in a solution or hybridized with the biomarker immobilized on the substrate and mRNA or cDNA extracted from a diagnostic object to confirm whether mRNA or cDNA of the same sequence is expressed.

Therefore, a biomarker used in the kit requires a process of removing one strand from its base sequence.

Refers to any substrate to which the biomarker can be coupled under conditions where the substrate retains the hybridization property and the background level of hybridization is kept low when the biomarker is in the form of a microarray immobilized on a substrate. Typically, the substrate can be a microtiter plate, a membrane (e.g., nylon or nitrocellulose) or a microsphere (bead) or chip.

Before application or fixation to the membrane, the biomarker may be modified to immobilize or improve the hybridization efficiency. Such modifications may include coupling with different reactive functional groups such as homopolymer tailings, aliphatic groups, NH2 groups, SH groups and carboxyl groups, or coupling with biotin, haptens or proteins.

"Hybridization" means a process in which two complementary strands of a nucleic acid are combined to form a double stranded molecule (hybrid).

The hybridization solution is a buffer solution that allows hybridization of the biomarker and the mRNA or cDNA extracted from the diagnostic object, and a solution known in the art can be used.

The kit may further include a detector capable of detecting a nucleic acid of a diagnostic object hybridized with the biomarker. The detector may be a scanner, a spectrophotometer, a liquid scintillation counter, or the like, but is not limited thereto. The kit of the present invention may further include a user's manual describing optimal reaction performance conditions.

The kit can qualitatively detect the progress of aging by comparing the standard outbreak amount of the biomarker in the same species as the diagnostic subject and the measured biomarker expression amount in the diagnostic subject. Specifically, Of the biomarker of the present invention and the amount of expression of the biomarker measured in the diagnostic individual are compared with each other, it is accurately determined that the amount of expression in the diagnostic subject is decreased, It can be quickly identified.

Another aspect of the present invention relates to an obesity-determining kit comprising the biomarker and the hybridization solution.

The biomarker is as described above, and may be stored in a solution, or the biomarker may be immobilized on the substrate at a high density. In other words, the biomarkers may be in a microarray form immobilized in a predetermined region. Such microarrays are well known in the art.

The kit may be dispersed in a solution or hybridized with the biomarker immobilized on the substrate and mRNA or cDNA extracted from a diagnostic object to confirm whether mRNA or cDNA of the same sequence is expressed.

Therefore, a biomarker used in the kit requires a process of removing one strand from its base sequence.

Refers to any substrate to which the biomarker can be coupled under conditions where the substrate retains the hybridization property and the background level of hybridization is kept low when the biomarker is in the form of a microarray immobilized on a substrate. Typically, the substrate can be a microtiter plate, a membrane (e.g., nylon or nitrocellulose) or a microsphere (bead) or chip.

Before application or fixation to the membrane, the biomarker may be modified to immobilize or improve the hybridization efficiency. Such modifications may include coupling with different reactive functional groups such as homopolymer tailings, aliphatic groups, NH2 groups, SH groups and carboxyl groups, or coupling with biotin, haptens or proteins.

"Hybridization" means a process in which two complementary strands of a nucleic acid are combined to form a double stranded molecule (hybrid).

The hybridization solution is a buffer solution that allows hybridization of the biomarker and the mRNA or cDNA extracted from the diagnostic object, and a solution known in the art can be used.

The kit may further include a detector capable of detecting a nucleic acid of a diagnostic object hybridized with the biomarker. The detector may be a scanner, a spectrophotometer, a liquid scintillation counter, or the like, but is not limited thereto. The kit of the present invention may further include a user's manual describing optimal reaction performance conditions.

The kit can qualitatively detect the degree of obesity by comparing the standard amount of biomarker of the same species as the diagnostic individual and the measured biomarker expression amount in the diagnostic individual. Specifically, When the expression level of the biomarker is compared with the standard expression level of the biomarker and the expression level of the biomarker measured in the diagnostic subject, if the expression level in the diagnostic subject is decreased, the diagnostic subject has a higher average triglyceride content and liposome size and density State can be accurately and quickly determined.

Another aspect of the present invention relates to a cancer diagnostic kit comprising the biomarker and the hybridization solution.

The biomarker is as described above, and may be stored in a solution, or the biomarker may be immobilized on the substrate at a high density. In other words, the biomarkers may be in a microarray form immobilized in a predetermined region. Such microarrays are well known in the art.

The kit may be dispersed in a solution or hybridized with the biomarker immobilized on the substrate and mRNA or cDNA extracted from a diagnostic object to confirm whether mRNA or cDNA of the same sequence is expressed.

Therefore, a biomarker used in the kit requires a process of removing one strand from its base sequence.

Refers to any substrate to which the biomarker can be coupled under conditions where the substrate retains the hybridization property and the background level of hybridization is kept low when the biomarker is in the form of a microarray immobilized on a substrate. Typically, the substrate can be a microtiter plate, a membrane (e.g., nylon or nitrocellulose) or a microsphere (bead) or chip.

Before application or fixation to the membrane, the biomarker may be modified to immobilize or improve the hybridization efficiency. Such modifications may include coupling with different reactive functional groups such as homopolymer tailings, aliphatic groups, NH2 groups, SH groups and carboxyl groups, or coupling with biotin, haptens or proteins.

"Hybridization" means a process in which two complementary strands of a nucleic acid are combined to form a double stranded molecule (hybrid).

The hybridization solution is a buffer solution that allows hybridization of the biomarker and the mRNA or cDNA extracted from the diagnostic object, and a solution known in the art can be used.

The kit may further include a detector capable of detecting a nucleic acid of a diagnostic object hybridized with the biomarker. The detector may be a scanner, a spectrophotometer, a liquid scintillation counter, or the like, but is not limited thereto. The kit of the present invention may further include a user's manual describing optimal reaction performance conditions.

The kit can qualitatively detect the degree of obesity by comparing the standard amount of biomarker of the same species as the diagnostic individual and the measured biomarker expression amount in the diagnostic individual. Specifically, When the standard expression level of the biomarker is compared with the expression level of the biomarker measured in the diagnostic subject, if the expression level in the diagnostic subject is decreased, the diagnostic subject can accurately and promptly show that the cancer develops or grows Can be diagnosed.

In addition, the kit according to the present invention can be used for aging progress discrimination, obesity discrimination and cancer diagnosis as described above, but can also be used as a complex diagnosis kit for simultaneously detecting aging progress, obesity increase, and cancer occurrence .

As described above, the complex diagnostic kit comprises any one of the nucleotide sequences selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, a complementary base sequence thereof, And a hybridization solution containing at least one of the nucleotide sequences selected from the group consisting of SEQ ID NO: 1 and SEQ ID NO: 2. As the expression of the biomarker is decreased, the aging of the Drosophila mutant controlling specific gene expression using the UAS- , The increase in fat pore size and size in the triglyceride and fat body tissues, and the decrease in cancer incidence, the standard outbreak of biomarker in the same species as the diagnostic individual and the biomarker expression in the diagnostic individual , Which can simultaneously detect progress of aging, increase in obesity and cancer occurrence, It can be used as a single-use kit.

Another aspect of the present invention relates to a method of determining aging progress comprising the following steps.

I) isolating and extracting RNA from a diagnostic individual;

II) contacting the isolated RNA or cDNA synthesized therefrom with the aging-progression-determining kit to hybridize the RNA or cDNA with the biomarker; And

III) detecting the degree of hybridization between the biomarker and the RNA or cDNA.

A method known in the art can be used as a method for separating RNA from the diagnostic entity.

Specifically, it may be a step of isolating RNA from the cells of the separated diagnostic subject in vitro using cells separated from the diagnostic subject.

According to an embodiment of the present invention, the cDNA may be a first strand cDNA synthesized using the separated RNA as a template. The first strand cDNA can be synthesized by a method commonly used in the art, for example, a reverse transcriptase, an RNase block ribonuclease inhibitor, or the like. Examples of reverse transcriptase enzymes include, but are not limited to, reverse transcriptase from a variety of sources such as avian myeloblastosis virus-derived virus reverse transcriptase (AMV RTase), mouse leukemia virus-derived reverse transcriptase murine leukemia virus-derived virus reverse transcriptase (MMLV RTase) and Rous-associated virus 2 reverse transcriptase (RAV-2 RTase).

Preferably, the cDNA can be labeled with a detectable labeling substance. The labeling substance may be a fluorescent, phosphorescent or radioactive substance, but is not limited thereto. Preferably, the labeling substance is Cy5 or Cy3. When the first strand cDNA is synthesized by labeling Cy5 or Cy3 at the 5'-end of the primer, the target sequence may be labeled with a detectable fluorescent labeling substance. When the radioactive isotope such as ³² P or ³⁵ S is added to the reaction solution during the synthesis of the first strand cDNA, the synthesis product is synthesized and the radioactive is incorporated into the synthesis product and the synthesis product is labeled with radioactivity have.

The step of detecting the degree of hybridization may be performed through capillary electrophoresis, gel electrophoresis, radioactivity measurement, fluorescence measurement or phosphorescence measurement.

According to another embodiment of the present invention, the aging progress discrimination method may further include a step of comparing aging progress of the diagnostic subject by comparing the detection result with the standard of the diagnostic subject.

On the other hand, the aging progression discrimination method is for providing information necessary for discriminating the aging progress of the diagnostic subject, separating RNA from the cells of the separated diagnostic subject in vitro using the cells isolated from the diagnostic subject, A probe capable of detecting the expression of any one of the nucleotide sequences selected from the group consisting of SEQ ID NOS: 1 to 4 or the expression of any one of the amino acid sequences selected from the group consisting of SEQ ID NOS: 5 to 8 1 to 4 or a protein comprising any one of the amino acid sequences selected from the group consisting of SEQ ID NOS: 5 to 8 is detected by the addition of an antibody having an amino acid sequence selected from the group consisting of SEQ ID NOS: .

The method may further include comparing the detected expression level of the gene and protein with the standard of the diagnostic subject to determine whether or not the diagnostic subject is progressing to aging.

The diagnostic object may be a mammal or an insect other than human or human.

Another aspect of the present invention relates to an obesity discrimination method comprising the following steps.

I) isolating and extracting RNA from a diagnostic individual;

II) contacting the isolated RNA or the synthesized cDNA with the obesity-determining kit to hybridize the RNA or cDNA with the biomarker; And

III) detecting the degree of hybridization between the biomarker and the RNA or cDNA.

A method known in the art can be used as a method for separating RNA from the diagnostic entity.

Specifically, it may be a step of isolating RNA from the cells of the separated diagnostic subject in vitro using cells separated from the diagnostic subject.

According to an embodiment of the present invention, the cDNA may be a first strand cDNA synthesized using the separated RNA as a template. The first strand cDNA can be synthesized by a method commonly used in the art, for example, a reverse transcriptase, an RNase block ribonuclease inhibitor, or the like. Examples of reverse transcriptase enzymes include, but are not limited to, reverse transcriptase from a variety of sources such as avian myeloblastosis virus-derived virus reverse transcriptase (AMV RTase), mouse leukemia virus-derived reverse transcriptase murine leukemia virus-derived virus reverse transcriptase (MMLV RTase) and Rous-associated virus 2 reverse transcriptase (RAV-2 RTase).

Preferably, the cDNA can be labeled with a detectable labeling substance. The labeling substance may be a fluorescent, phosphorescent or radioactive substance, but is not limited thereto. Preferably, the labeling substance is Cy5 or Cy3. When the first strand cDNA is synthesized by labeling Cy5 or Cy3 at the 5'-end of the primer, the target sequence may be labeled with a detectable fluorescent labeling substance. When the radioactive isotope such as ³² P or ³⁵ S is added to the reaction solution during the synthesis of the first strand cDNA, the synthesis product is synthesized and the radioactive is incorporated into the synthesis product and the synthesis product is labeled with radioactivity have.

The step of detecting the degree of hybridization may be performed through capillary electrophoresis, gel electrophoresis, radioactivity measurement, fluorescence measurement or phosphorescence measurement.

According to another embodiment of the present invention, the above-mentioned method of discriminating obesity comprises comparing the detection result with the standard of the diagnostic object, thereby determining an increase in the triglyceride content of the diagnostic subject, Based on the result of the determination.

On the other hand, the above-mentioned obesity discrimination method is for providing information necessary for discrimination of obesity of the diagnosis object, and separating RNA from the cells of the separated diagnosis subject in vitro using the cells isolated from the diagnosis subject, A probe capable of detecting the expression of any one of the nucleotide sequences selected from the group consisting of SEQ ID NOS: 1 to 4 or an antibody capable of detecting the expression of any one of the amino acid sequences selected from the group consisting of SEQ ID NOS: 5 to 8 1 to 4 or a protein comprising any one of the amino acid sequences selected from the group consisting of SEQ ID NOS: 5 to 8 by the addition of the nucleotide sequence of SEQ ID NO: .

The step of comparing the detected gene and protein expression level with the standard of the diagnostic subject to determine whether the diagnostic subject is obese (increase in the triglyceride content and increase in the size and density of the fat sphere in the fat body tissue) .

The diagnostic object may be a mammal or an insect other than human or human.

Another aspect of the invention relates to a method of diagnosing cancer comprising the steps of:

I) isolating and extracting RNA from a diagnostic individual;

II) contacting the isolated RNA or the synthesized cDNA with the cancer diagnostic kit to hybridize the RNA or cDNA with the biomarker; And

III) detecting the degree of hybridization between the biomarker and the RNA or cDNA.

A method known in the art can be used as a method for separating RNA from the diagnostic entity.

Specifically, it may be a step of isolating RNA from the cells of the separated diagnostic subject in vitro using cells separated from the diagnostic subject.

According to an embodiment of the present invention, the cDNA may be a first strand cDNA synthesized using the separated RNA as a template. The first strand cDNA can be synthesized by a method commonly used in the art, for example, a reverse transcriptase, an RNase block ribonuclease inhibitor, or the like. Examples of reverse transcriptase enzymes include, but are not limited to, reverse transcriptase from a variety of sources such as avian myeloblastosis virus-derived virus reverse transcriptase (AMV RTase), mouse leukemia virus-derived reverse transcriptase murine leukemia virus-derived virus reverse transcriptase (MMLV RTase) and Rous-associated virus 2 reverse transcriptase (RAV-2 RTase).

Preferably, the cDNA can be labeled with a detectable labeling substance. The labeling substance may be a fluorescent, phosphorescent or radioactive substance, but is not limited thereto. Preferably, the labeling substance is Cy5 or Cy3. When the first strand cDNA is synthesized by labeling Cy5 or Cy3 at the 5'-end of the primer, the target sequence may be labeled with a detectable fluorescent labeling substance. When the radioactive isotope such as ³² P or ³⁵ S is added to the reaction solution during the synthesis of the first strand cDNA, the synthesis product is synthesized and the radioactive is incorporated into the synthesis product and the synthesis product is labeled with radioactivity have.

The step of detecting the degree of hybridization may be performed through capillary electrophoresis, gel electrophoresis, radioactivity measurement, fluorescence measurement or phosphorescence measurement.

According to another embodiment of the present invention, the cancer diagnosis method may further include a step of comparing the detection result with the standard of the diagnostic object to diagnose cancer through the cancer occurrence or growth of the diagnostic object.

On the other hand, the cancer diagnosis method is for providing information necessary for cancer diagnosis of the diagnostic individual, separating RNA from the cells of the separated diagnostic individual in vitro using the cells isolated from the diagnostic individual, A probe capable of detecting the expression of any one of the nucleotide sequences selected from the group consisting of SEQ ID NOS: 1 to 4 or an antibody capable of detecting the expression of any one of the amino acid sequences selected from the group consisting of SEQ ID NOS: 5 to 8 1 to 4 or a protein comprising any one of the amino acid sequences selected from the group consisting of SEQ ID NOS: 5 to 8 by the addition of the nucleotide sequence of SEQ ID NO: .

The method may further include the step of diagnosing cancer occurrence and growth of the diagnostic subject by comparing the detected gene and protein expression level with the standard of the diagnostic subject.

The diagnostic object may be a mammal or an insect other than human or human.

In the present invention, it is possible to simultaneously detect progress of aging, increase in obesity, and whether or not cancer has occurred by using the above-described multiple diagnosis kit according to the present invention. It is possible.

As described above, the complex diagnosis kit is performed in the same manner as the determination method using each of the above-described kits.

However, the detection result can be compared with the standard of the diagnostic subject to determine whether the diagnosis subject is aged, obese, cancerous, or growing.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

Example

<Preparation process>

In order to investigate the progression of aging, we first examined whether Actin-GS-Gal4 Drosophila and wild-type (w1118) Drosophila were crossed to affect the lifespan of RU486. As a result, RU486 did not affect the lifespan (See Figure 1 below).

Actin-GS-Gal4 is expressed in Drosophila in the presence of RU486, and the expression of UAS-fbp RNAi is reduced by the transcription of fbp (Fructose-1,6-bisphosphatase, CG31692) . The amount of fbp mRNA was measured by RT-PCR to confirm the expression of fbp in the offspring (F1) fruit flies obtained by crossing Actin-GS-Gal4 with UAS-fbp RNAi fruit flies. Was significantly reduced compared to when it did not feed RU486. This result demonstrates that Actin-GS-Gal4 and UAS-fbp RNAi are functioning normally (see FIG. 2 below).

[Example 1: Actin-GS-Gal4 > RT-PCR of UAS-fbp RNAi]

To confirm that UAS-fbp RNAi Drosophila is functioning properly, Actin-GS-Gal4 Drosophila females and UAS-fbp RNAi Drosophila males were mated to collect F1 males of F1 generation and cultured in media containing RU486 (150 μM) , Agar 0.7%, corn flour 6.1%, yeast 2.6%, 10% Tegosept 1.6%, water 80.7%) and RU486 were grown for 10 days and then the reverse transcriptase polymerase chain reaction (Reverse transcriptase mediated PCR, RT-PCR). As a result, it was confirmed that the amount of mRNA of fbp in the cultured Drosophila melanogaster was decreased statistically by the UAS-fbp RNAi in the medium containing RU486.

[Example 2: Actin-GS-Gal4 fed with RU486> Life test of UAS-fbp RNAi Drosophila]

Actin-GS-Gal4 Drosophila females and UAS-fbp RNAi Drosophila males were mated to collect males of F1 generation and 120 rats were placed in a medium containing RU486 (150 μM) and RU486 medium. The lifespan was measured while growing at 25 ° C, 50% relative humidity, and 12:12 hour light cycle. The experiment measuring the lifetime was repeated three times. As shown in the results of FIG. 3, it can be seen that when the expression of fbp is reduced, the life span decreases, that is, the aging progresses further (see FIG. 3).

[Example 3: Actin-GS-Gal4 fed with RU486] Measurement of Neutral Fat in UAS-fbp RNAi Drosophila]

Actin-GS-Gal4 Drosophila females and UAS-fbp RNAi Drosophila males were mated to collect males of F1 generation, and about 100 fleas were added to a medium containing RU486 (150 μM) and RU486 The neutral fat was measured using thin layer chromatography (TLC) (see FIG. 4 below as a result of measurement of the change in the triglyceride content of wild type Drosophila fed with RU486). 10 μl of Drosophila was placed in an Eppendorf tube, and 100 μl of an extraction solvent (10 mM Tris, 1 mM EDTA, 0.1% TritonX-100) was added for 5 minutes and centrifuged. 5 μl of the supernatant was subjected to TLC And then dropped on a plate. TLC eluting solvent composition for the separation of triglyceride was hexane; diethylether; acetic acid (70: 30: 1) and sesame oil (Sigma-Aldrich S3547-250ML). After developing on the TLC plate, the TLC plate was dried, placed in an iodine saturation box, stained with TLC plates, pictures were taken, and neutral fat bands were quantified using the Image J program. As a result, it was confirmed that the content of triglyceride was significantly increased in the group in which the expression of fbp was inhibited in the fruit fly fed with RU486 (see FIG. 5).

[Example 4: Analysis of fat-confocal microscope of Actin-GS-Gal4 > UAS-fbp RNAi fruit flies fed with RU486]

Actin-GS-Gal4 Drosophila females and UAS-fbp RNAi Drosophila males were mated to observe lipid size, and the males of F1 generation were collected and cultured for 10 days in a medium containing RU486 (150 μM) and a medium without RU486 And dyed with Nile red. Drosophila was dissected and fixed in 4% paraformaldehyde solution dissolved in PBS for 30 minutes at room temperature. After washing with PBS, 0.5 mg / ml Nile red (Sigma) solution was diluted 1: 2,500 and incubated at room temperature for 30 minutes After staining, wash with distilled water twice. The dyed samples were then placed in a 80% glycerol solution on a slide glass and measured by a confocal microscope (see FIG. 6, below). Actin-GS-Gal4 / UAS-nls. GFP fed with RU486 Focus confocal microscope photo). As a result, it was confirmed that the size and density of lipids were increased (see FIG. 7).

[Example 5: preparation of cancer proliferation model fruit fly (UAS-Ras85D; c765-Gal4)] [

In order to carry out a study on cancer, the cancer growth model flies were prepared by crossing c765-Gal4 Drosophila and UAS-Ras85D Drosophila.

[Example 6: phenotypic analysis of cancer proliferation model flies (UAS-Ras85D; c765-Gal4)

Manufactured UAS-Ras85D; C765-Gal4 / + CS10, UAS-Ras85D / + CS10 and UAS-Ras85D / + CS10, which crossed wild-type flies (CS10) with c765-Gal4 and CS10 to analyze the phenotype of the c765-Gal4 cancer growth blackfly flies model. As a result of comparing the blade lengths of c765-Gal4 / + CS10, UAS-Ras85D / + CS10; The wing length of c765-Gal4 / + CS10 was longer than that of the three control groups. The wing length was compared with the length of the chest to compensate for the difference between the individual flies. That is, cancer proliferation assay model. (See Figure 8 below)

[Example 7: Influence of inhibition of fbp gene expression on phenotype in cancer proliferation model fruit fly (UAS-Ras85D; c765-Gal4)] [

In order to estimate the effect of inhibition of fbp gene expression on cancer growth, the wing phenotype of the F1 generation obtained by crossing the cancer growth model flies (UAS-Ras85D; c765-Gal4) with UAS-fbp RNAi Drosophila was investigated. As a result, it was confirmed that the wing length and area increased due to overexpression of Ras85D were significantly decreased when the expression of fbp was inhibited (see FIGS. 9 and 10 ').

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims. It is natural.

<110> Korea University Research and Business Foundation <120> Fbp gene common associating life-reduction, cancer and obesity <130> HPC-6032 <160> 8 <170> KoPatentin 3.0 <210> 1 <211> 1005 <212> DNA <213> Drosophila sp. <400> 1 atgacccaac agaggccagc tttcgactcc aatgcgatga cgctgacgcg tttcgtgctg 60 caggagcagc gaaagttcaa gagcgccact ggcgatctct cccagctgct caactccatc 120 cagaccgcca tcaaggctac atcatccgcg gtgcggaagg caggtatcgc caagctccat 180 ggattcgctg gcgacgtgaa tgtccaaggc gaggaggtca agaaactgga cgtgctctcc 240 aacgagctgt tcatcaacat gctgaagtca tcctatacca catgtctaat ggtttccgag 300 gagaacgaga atgtgatcga ggtggaagtg gagaaacagg gcaaatacat cgtgtgcttc 360 gatcccttgg atggatcctc caacatagac tgcctggtgt cgatcggttc aatcttcgcc 420 atttaccgca agaaaagcga tggtccgccc acagtggagg atgcactgca gcccggaaat 480 cagctggtgg ccgccggcta cgcgctatac ggttcggcca cagcaattgt cctgggtctg 540 ggttcgggag tgaatggctt cacttatgac ccggccatcg gagagttcgt gctgaccgat 600 cccaacatgc gggtgccgga gaagggaaag atatactcta tcaacgaggg atatgcagcg 660 gattgggagg atggtgtctt caactacatt gcggccaaga aggatcccgc caagggaaag 720 ccctatggag cgcggtacgt gggttccatg gtcgcggatg tgcatcgcac cattaaatac 780 ggcggcatct ttatctatcc ggcaacaaag tccgctccca gcggaaaact tcgtctgctg 840 tacgagtgcg tgcccatggc ctatctgatg atccaggctg gaggtctggc cagcgacgga 900 aagatcagca ttttggacat tgtgcccaag aagatccacg agcgcagtcc catattccta 960 ggatccaagt ccgacgtgga ggaggcactt agctacttaa agtga 1005 <210> 2 <211> 969 <212> DNA <213> Drosophila sp. <400> 2 atgacgctga cgcgtttcgt gctgcaggag cagcgaaagt tcaagagcgc cactggcgat 60 ctctcccagc tgctcaactc catccagacc gccatcaagg ctacatcatc cgcggtgcgg 120 aaggcaggta tcgccaagct ccatggattc gctggcgacg tgaatgtcca aggcgaggag 180 gtcaagaaac tggacgtgct ctccaacgag ctgttcatca acatgctgaa gtcatcctat 240 accacatgtc taatggtttc cgaggagaac gagaatgtga tcgaggtgga agtggagaaa 300 cagggcaaat acatcgtgtg cttcgatccc ttggatggat cctccaacat agactgcctg 360 gtgtcgatcg gttcaatctt cgccatttac cgcaagaaaa gcgatggtcc gcccacagtg 420 gaggatgcac tgcagcccgg aaatcagctg gtggccgccg gctacgcgct atacggttcg 480 gccacagcaa ttgtcctggg tctgggttcg ggagtgaatg gcttcactta tgacccggcc 540 atcggagagt tcgtgctgac cgatcccaac atgcgggtgc cggagaaggg aaagatatac 600 tctatcaacg agggatatgc agcggattgg gaggatggtg tcttcaacta cattgcggcc 660 aagaaggatc ccgccaaggg aaagccctat ggagcgcggt acgtgggttc catggtcgcg 720 gatgtgcatc gcaccattaa atacggcggc atctttatct atccggcaac aaagtccgct 780 cccagcggaa aacttcgtct gctgtacgag tgcgtgccca tggcctatct gatgatccag 840 gctggaggtc tggccagcga cggaaagatc agcattttgg acattgtgcc caagaagatc 900 cacgagcgca gtcccatatt cctaggatcc aagtccgacg tggaggaggc acttagctac 960 ttaaagtga 969 <210> 3 <211> 969 <212> DNA <213> Drosophila sp. <400> 3 atgacgctga cgcgtttcgt gctgcaggag cagcgaaagt tcaagagcgc cactggcgat 60 ctctcccagc tgctcaactc catccagacc gccatcaagg ctacatcatc cgcggtgcgg 120 aaggcaggta tcgccaagct ccatggattc gctggcgacg tgaatgtcca aggcgaggag 180 gtcaagaaac tggacgtgct ctccaacgag ctgttcatca acatgctgaa gtcatcctat 240 accacatgtc taatggtttc cgaggagaac gagaatgtga tcgaggtgga agtggagaaa 300 cagggcaaat acatcgtgtg cttcgatccc ttggatggat cctccaacat agactgcctg 360 gtgtcgatcg gttcaatctt cgccatttac cgcaagaaaa gcgatggtcc gcccacagtg 420 gaggatgcac tgcagcccgg aaatcagctg gtggccgccg gctacgcgct atacggttcg 480 gccacagcaa ttgtcctggg tctgggttcg ggagtgaatg gcttcactta tgacccggcc 540 atcggagagt tcgtgctgac cgatcccaac atgcgggtgc cggagaaggg aaagatatac 600 tctatcaacg agggatatgc agcggattgg gaggatggtg tcttcaacta cattgcggcc 660 aagaaggatc ccgccaaggg aaagccctat ggagcgcggt acgtgggttc catggtcgcg 720 gatgtgcatc gcaccattaa atacggcggc atctttatct atccggcaac aaagtccgct 780 cccagcggaa aacttcgtct gctgtacgag tgcgtgccca tggcctatct gatgatccag 840 gctggaggtc tggccagcga cggaaagatc agcattttgg acattgtgcc caagaagatc 900 cacgagcgca gtcccatatt cctaggatcc aagtccgacg tggaggaggc acttagctac 960 ttaaagtga 969 <210> 4 <211> 1032 <212> DNA <213> Drosophila sp. <400> 4 atggcggcca gtagcggtga ttccaaaatg acccaacaga ggccagcttt cgactccaat 60 gcgatgacgc tgacgcgttt cgtgctgcag gagcagcgaa agttcaagag cgccactggc 120 gatctctccc agctgctcaa ctccatccag accgccatca aggctacatc atccgcggtg 180 cggaaggcag gtatcgccaa gctccatgga ttcgctggcg acgtgaatgt ccaaggcgag 240 gaggtcaaga aactggacgt gctctccaac gagctgttca tcaacatgct gaagtcatcc 300 tataccacat gtctaatggt ttccgaggag aacgagaatg tgatcgaggt ggaagtggag 360 aaacagggca aatacatcgt gtgcttcgat cccttggatg gatcctccaa catagactgc 420 ctggtgtcga tcggttcaat cttcgccatt taccgcaaga aaagcgatgg tccgcccaca 480 gtggaggatg cactgcagcc cggaaatcag ctggtggccg ccggctacgc gctatacggt 540 tcggccacag caattgtcct gggtctgggt tcgggagtga atggcttcac ttatgacccg 600 gccatcggag agttcgtgct gaccgatccc aacatgcggg tgccggagaa gggaaagata 660 tactctca acgagggata tgcagcggat tgggaggatg gtgtcttcaa ctacattgcg 720 gccaagaagg atcccgccaa gggaaagccc tatggagcgc ggtacgtggg ttccatggtc 780 gcggatgtgc atcgcaccat taaatacggc ggcatcttta tctatccggc aacaaagtcc 840 gctcccagcg gaaaacttcg tctgctgtac gagtgcgtgc ccatggccta tctgatgatc 900 caggctggag gtctggccag cgacggaaag atcagcattt tggacattgt gcccaagaag 960 atccacgagc gcagtcccat attcctagga tccaagtccg acgtggagga ggcacttagc 1020 tacttaaagt ga 1032 <210> 5 <211> 334 <212> PRT <213> Drosophila sp. <400> 5 Met Thr Gln Gln Arg Pro Ala Phe Asp Ser Asn Ala Met Thr Leu Thr   1 5 10 15 Arg Phe Val Leu Gln Glu Gln Arg Lys Phe Lys Ser Ala Thr Gly Asp              20 25 30 Leu Ser Gln Leu Leu Asn Ser Ile Gln Thr Ala Ile Lys Ala Thr Ser          35 40 45 Ser Ala Val Arg Lys Ala Gly Ile Ala Lys Leu His Gly Phe Ala Gly      50 55 60 Asp Val Asn Val Gln Gly Glu Glu Val Lys Lys Leu Asp Val Leu Ser  65 70 75 80 Asn Glu Leu Phe Ile Asn Met Leu Lys Ser Ser Tyr Thr Thr Cys Leu                  85 90 95 Met Val Ser Glu Glu Asn Glu Asn Val Glu Val Glu Val Glu Lys             100 105 110 Gln Gly Lys Tyr Ile Val Cys Phe Asp Pro Leu Asp Gly Ser Ser Asn         115 120 125 Ile Asp Cys Leu Val Ser Ile Gly Ser Ile Phe Ala Ile Tyr Arg Lys     130 135 140 Lys Ser Asp Gly Pro Pro Thr Val Glu Asp Ala Leu Gln Pro Gly Asn 145 150 155 160 Gln Leu Val Ala Gly Tyr Ala Leu Tyr Gly Ser Ala Thr Ala Ile                 165 170 175 Val Leu Gly Leu Gly Ser Gly Val Asn Gly Phe Thr Tyr Asp Pro Ala             180 185 190 Ile Gly Glu Phe Val Leu Thr Asp Pro Asn Met Arg Val Pro Glu Lys         195 200 205 Gly Lys Ile Tyr Ser Ile Asn Glu Gly Tyr Ala Ala Asp Trp Glu Asp     210 215 220 Gly Val Phe Asn Tyr Ile Ala Ala Lys Lys Asp Pro Ala Lys Gly Lys 225 230 235 240 Pro Tyr Gly Ala Arg Tyr Val Gly Ser Met Val Ala Asp Val His Arg                 245 250 255 Thr Ile Lys Tyr Gly Gly Ile Phe Ile Tyr Pro Ala Thr Lys Ser Ala             260 265 270 Pro Ser Gly Lys Leu Arg Leu Leu Tyr Glu Cys Val Pro Met Ala Tyr         275 280 285 Leu Met Ile Gln Ala Gly Gly Leu Ala Ser Asp Gly Lys Ile Ser Ile     290 295 300 Leu Asp Ile Val Pro Lys Lys Ile His Glu Arg Ser Pro Ile Phe Leu 305 310 315 320 Gly Ser Lys Ser Asp Val Glu Glu Ala Leu Ser Tyr Leu Lys                 325 330 <210> 6 <211> 322 <212> PRT <213> Drosophila sp. <400> 6 Met Thr Leu Thr Arg Phe Val Leu Gln Glu Gln Arg Lys Phe Lys Ser   1 5 10 15 Ala Thr Gly Asp Leu Ser Gln Leu Leu Asn Ser Ile Gln Thr Ala Ile              20 25 30 Lys Ala Thr Ser Ser Ala Val Arg Lys Ala Gly Ile Ala Lys Leu His          35 40 45 Gly Phe Ala Gly Asp Val Asn Val Gln Gly Glu Glu Val Lys Lys Leu      50 55 60 Asp Val Leu Ser Asn Glu Leu Phe Ile Asn Met Leu Lys Ser Ser Tyr  65 70 75 80 Thr Cys Leu Met Val Ser Glu Glu Asn Glu Asn Val Ile Glu Val                  85 90 95 Glu Val Glu Lys Gln Gly Lys Tyr Ile Val Cys Phe Asp Pro Leu Asp             100 105 110 Gly Ser Ser Asn Ile Asp Cys Leu Val Ser Ile Gly Ser Ile Phe Ala         115 120 125 Ile Tyr Arg Lys Lys Ser Asp Gly Pro Pro Thr Val Glu Asp Ala Leu     130 135 140 Gln Pro Gly Asn Gln Leu Val Ala Gly Tyr Ala Leu Tyr Gly Ser 145 150 155 160 Ala Thr Ala Ile Val Leu Gly Leu Gly Ser Gly Val Asn Gly Phe Thr                 165 170 175 Tyr Asp Pro Ala Ile Gly Glu Phe Val Leu Thr Asp Pro Asn Met Arg             180 185 190 Val Pro Glu Lys Gly Lys Ile Tyr Ser Ile Asn Glu Gly Tyr Ala Ala         195 200 205 Asp Trp Glu Asp Gly Val Phe Asn Tyr Ile Ala Ala Lys Lys Asp Pro     210 215 220 Ala Lys Gly Lys Pro Tyr Gly Ala Arg Tyr Val Gly Ser Met Val Ala 225 230 235 240 Asp Val His Arg Thr Ile Lys Tyr Gly Gly Ile Phe Ile Tyr Pro Ala                 245 250 255 Thr Lys Ser Ala Pro Ser Gly Lys Leu Arg Leu Leu Tyr Glu Cys Val             260 265 270 Pro Met Ala Tyr Leu Met Ile Gln Ala Gly Gly Leu Ala Ser Asp Gly         275 280 285 Lys Ile Ser Ile Leu Asp Ile Val Pro Lys Lys Ile His Glu Arg Ser     290 295 300 Pro Ile Phe Leu Gly Ser Lys Ser Asp Val Glu Glu Ala Leu Ser Tyr 305 310 315 320 Leu Lys         <210> 7 <211> 322 <212> PRT <213> Drosophila sp. <400> 7 Met Thr Leu Thr Arg Phe Val Leu Gln Glu Gln Arg Lys Phe Lys Ser   1 5 10 15 Ala Thr Gly Asp Leu Ser Gln Leu Leu Asn Ser Ile Gln Thr Ala Ile              20 25 30 Lys Ala Thr Ser Ser Ala Val Arg Lys Ala Gly Ile Ala Lys Leu His          35 40 45 Gly Phe Ala Gly Asp Val Asn Val Gln Gly Glu Glu Val Lys Lys Leu      50 55 60 Asp Val Leu Ser Asn Glu Leu Phe Ile Asn Met Leu Lys Ser Ser Tyr  65 70 75 80 Thr Cys Leu Met Val Ser Glu Glu Asn Glu Asn Val Ile Glu Val                  85 90 95 Glu Val Glu Lys Gln Gly Lys Tyr Ile Val Cys Phe Asp Pro Leu Asp             100 105 110 Gly Ser Ser Asn Ile Asp Cys Leu Val Ser Ile Gly Ser Ile Phe Ala         115 120 125 Ile Tyr Arg Lys Lys Ser Asp Gly Pro Pro Thr Val Glu Asp Ala Leu     130 135 140 Gln Pro Gly Asn Gln Leu Val Ala Gly Tyr Ala Leu Tyr Gly Ser 145 150 155 160 Ala Thr Ala Ile Val Leu Gly Leu Gly Ser Gly Val Asn Gly Phe Thr                 165 170 175 Tyr Asp Pro Ala Ile Gly Glu Phe Val Leu Thr Asp Pro Asn Met Arg             180 185 190 Val Pro Glu Lys Gly Lys Ile Tyr Ser Ile Asn Glu Gly Tyr Ala Ala         195 200 205 Asp Trp Glu Asp Gly Val Phe Asn Tyr Ile Ala Ala Lys Lys Asp Pro     210 215 220 Ala Lys Gly Lys Pro Tyr Gly Ala Arg Tyr Val Gly Ser Met Val Ala 225 230 235 240 Asp Val His Arg Thr Ile Lys Tyr Gly Gly Ile Phe Ile Tyr Pro Ala                 245 250 255 Thr Lys Ser Ala Pro Ser Gly Lys Leu Arg Leu Leu Tyr Glu Cys Val             260 265 270 Pro Met Ala Tyr Leu Met Ile Gln Ala Gly Gly Leu Ala Ser Asp Gly         275 280 285 Lys Ile Ser Ile Leu Asp Ile Val Pro Lys Lys Ile His Glu Arg Ser     290 295 300 Pro Ile Phe Leu Gly Ser Lys Ser Asp Val Glu Glu Ala Leu Ser Tyr 305 310 315 320 Leu Lys         <210> 8 <211> 343 <212> PRT <213> Drosophila sp. <400> 8 Met Ala Ala Ser Ser Gly Asp Ser Lys Met Thr Gln Gln Arg Pro Ala   1 5 10 15 Phe Asp Ser Asn Ala Met Thr Leu Thr Arg Phe Val Leu Gln Glu Gln              20 25 30 Arg Lys Phe Lys Ser Ala Thr Gly Asp Leu Ser Gln Leu Leu Asn Ser          35 40 45 Ile Gln Thr Ala Ile Lys Ala Thr Ser Ser Ala Val      50 55 60 Ile Ala Lys Leu His Gly Phe Ala Gly Asp Val Asn Val Gln Gly Glu  65 70 75 80 Glu Val Lys Lys Leu Asp Val Leu Ser Asn Glu Leu Phe Ile Asn Met                  85 90 95 Leu Lys Ser Ser Tyr Thr Thr Cys Leu Met Val Ser Glu Glu Asn Glu             100 105 110 Asn Val Ile Glu Val Glu Val Glu Lys Gln Gly Lys Tyr Ile Val Cys         115 120 125 Phe Asp Pro Leu Asp Gly Ser Ser Asn Ile Asp Cys Leu Val Ser Ile     130 135 140 Gly Ser Ile Phe Ala Ile Tyr Arg Lys Lys Ser Asp Gly Pro Pro Thr 145 150 155 160 Val Glu Asp Ala Leu Gln Pro Gly Asn Gln Leu Val Ala Ala Gly Tyr                 165 170 175 Ala Leu Tyr Gly Ser Ala Thr Ala Ile Val Leu Gly Leu Gly Ser Gly             180 185 190 Val Asn Gly Phe Thr Tyr Asp Pro Ala Ile Gly Glu Phe Val Leu Thr         195 200 205 Asp Pro Asn Met Arg Val Pro Glu Lys Gly Lys Ile Tyr Ser Ile Asn     210 215 220 Glu Gly Tyr Ala Ala Asp Trp Glu Asp Gly Val Phe Asn Tyr Ile Ala 225 230 235 240 Ala Lys Lys Asp Pro Ala Lys Gly Lys Pro Tyr Gly Ala Arg Tyr Val                 245 250 255 Gly Ser Met Val Ala Asp Val His Arg Thr Ile Lys Tyr Gly Gly Ile             260 265 270 Phe Ile Tyr Pro Ala Thr Lys Ser Ala Pro Ser Gly Lys Leu Arg Leu         275 280 285 Leu Tyr Glu Cys Val Pro Met Ala Tyr Leu Met Ile Gln Ala Gly Gly     290 295 300 Leu Ala Ser Asp Gly Lys Ile Ser Ile Leu Asp Ile Val Pro Lys Lys 305 310 315 320 Ile His Glu Arg Ser Pro Ile Phe Leu Gly Ser Lys Ser Asp Val Glu                 325 330 335 Glu Ala Leu Ser Tyr Leu Lys             340

Claims (17)

An aging process discrimination step comprising any one of a base sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, a complementary base sequence thereof and a base sequence consisting of these mRNAs Biomarker for. The method according to claim 1,
And aging of the diagnostic subject is promoted, thereby decreasing the expression amount of the biomarker. 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, a complementary base sequence thereof, and an mRNA of any of these nucleotide sequences. Biomarkers. The method of claim 3,
Wherein the amount of expression of the biomarker is decreased as the fat accumulation of the diagnostic subject is increased to increase obesity. 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, a complementary base sequence thereof, and an mRNA thereof, wherein the base sequence is any one selected from the group consisting of: Marker. 6. The method of claim 5,
Wherein the amount of expression of the biomarker is decreased as the number of cancer cells or cancer tissues of the diagnostic individual increases. A biomarker according to claim 1; And a hybridization solution. 8. The method of claim 7,
The biomarker may be present in a dispersed form in solution. Wherein the agar plate is present in the form of a microarray immobilized on a substrate. A biomarker according to claim 3; And a hybridization solution. 10. The method of claim 9,
The biomarker may be present in a dispersed form in solution. Wherein the oligonucleotide is present in the form of a microarray immobilized on a substrate. A biomarker according to claim 5; And a hybridization solution. 12. The method of claim 11,
The biomarker may be present in a dispersed form in solution. Wherein the microarray is present in the form of a microarray immobilized on a substrate. 6. A biomarker according to any one of claims 1 to 5; And a hybridization solution; and a diagnostic kit for simultaneous detection of aging progression discrimination, obesity discrimination and cancer diagnosis I) isolating and extracting RNA from a diagnostic individual;
II) contacting the separated RNA or a cDNA synthesized therefrom with the kit according to claim 7 to hybridize the RNA or cDNA with the biomarker; And
III) detecting the degree of hybridization between the biomarker and the RNA or cDNA. I) isolating and extracting RNA from a diagnostic individual;
II) contacting the separated RNA or a cDNA synthesized therefrom with the kit according to the above 9) to hybridize the RNA or cDNA with the biomarker; And
III) detecting the degree of hybridization between the biomarker and the RNA or cDNA. I) isolating and extracting RNA from a diagnostic individual;
II) contacting the separated RNA or a cDNA synthesized therefrom with the kit according to the above 11) to hybridize the RNA or cDNA with the biomarker; And
III) detecting the degree of hybridization between the biomarker and the RNA or cDNA. I) isolating and extracting RNA from a diagnostic individual;
II) contacting the isolated RNA or a cDNA synthesized therefrom with the kit according to the above 13) to hybridize the RNA or cDNA with the biomarker; And
III) detecting the hybridization degree between the biomarker and the RNA or cDNA, and detecting the aging progress discrimination, the obesity increase discrimination, and the cancer diagnosis.

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