KR20170012133A - biomarker comprising T3dh gene in common associating aging, cancer and obesity and diagnosis kit using thereof - Google Patents

Abstract

The present invention relates to a biomarker using a Type III alcohol dehydrogenase (T3dh, CG3425) gene mutually related to aging, obesity, and cancer. More specifically, the present invention relates to a diagnosing kit and a method using the gene. The gene and the diagnosing kit according to the present invention discover a T3dh 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 T3dh gene commonly associated with aging, obesity and cancer, and uses thereof, and biomarker comprising T3dh gene in common associating aging, cancer and obesity and diagnosis kit using thereof,

The present invention relates to a biomarker including T3dh (alcohol dehydrogenase type 3, type III alcohol dehydrogenase, CG3425) gene commonly associated with aging, obesity and cancer, and more particularly, to a biomarker including a diagnostic kit will be.

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, the incidence of cancer also increases. Interestingly, cancer-suppressing 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 achieve the above object, the present invention provides a biomarker for determining the progress of aging comprising the nucleotide sequence of SEQ ID NO: 1, a complementary base sequence thereof, and any one of the nucleotide sequences of mRNA thereof.

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

In order to accomplish the above object, the present invention provides a biomarker for obesity discrimination comprising a nucleotide sequence of SEQ ID NO: 1, a complementary base sequence thereof, and any one of nucleotide sequences consisting of mRNA thereof.

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 biomarker for cancer diagnosis comprising the nucleotide sequence of SEQ ID NO: 1, a complementary base sequence thereof, and any one of the nucleotide sequences of mRNAs thereof.

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 detecting progression of aging, discrimination of increase in obesity, and cancer diagnosis including the following steps.

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 degree of hybridization between the biomarker and the RNA or cDNA.

The biomarker and diagnostic kit according to the present invention finds a type III alcohol dehydrogenase (T3dh) gene commonly involved in aging, obesity and cancer, and analyzes the progress of aging, obesity and cancer . Thus, it becomes possible to comprehensively analyze or diagnose whether progress of aging of the diagnostic individual, cancer occurrence, and obesity occur.

Figure 1 is a graph showing Actin-GS-Gal4 / + W1118 lifetime curves for RU486 treatment.
Fig. 2 shows that the amount of T3dh mRNA decreased when Actin-GS-Gal4 and UAS-T3dh RNAi were used to inhibit the expression of T3dh.
FIG. 3 is a graph showing a result of confirming that the lifespan is shortened when the expression of T3dh decreases.
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-T3dh 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 fatty tissue of Actin-GS-Gal4 / UAS-T3dh RNAi fruit fly fed with RU486.
8 shows the results of comparing the blade lengths of cancer growth model flies (UAS-PI3K; c765-Gal4).
Fig. 9 shows the results of comparing the lengths of wings of cancer growth model flies (UAS-PI3K / +; c765-Gal4 / UAS-T3dh RNAi) inhibiting T3dh gene expression.
FIG. 10 shows the results of comparing the area of the wings of cancer growth model flies (UAS-PI3K / +; c765-Gal4 / UAS-T3dh RNAi) inhibiting T3dh gene expression.

Accordingly, the inventors of the present invention have made extensive efforts to find genes involved in aging, obesity and cancer. As a result, it has been found that T3dh (alcohol dehydrogenase type 3, type III alcohol dehydrogenase , CG3425) gene, thereby completing the present invention.

Hereinafter, the present invention will be described in more detail.

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 relates to a biomarker for determining the progress of aging comprising at least one base sequence selected from the group consisting of the nucleotide sequence of SEQ ID NO: 1, the complementary base sequence thereof and the mRNA thereof.

The nucleotide sequence of SEQ ID NO: 1, the complementary nucleotide sequence of the nucleotide sequence of SEQ ID NO: 1, and the mRNA of the nucleotide sequence of SEQ ID NO: 1 decreases the expression level 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 discriminating the progress of aging means a biomarker capable of qualitatively discriminating 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, by providing at least one nucleotide sequence selected from the group consisting of the nucleotide sequence of SEQ ID NO: 1 in the present invention, the complementary nucleotide sequence thereof, and the mRNA thereof as the biomarker for determining progress of aging, But can accurately and simply determine the aging progress of the diagnostic object.

The nucleotide sequence selected from the group consisting of SEQ ID NO: 1 is not particularly limited as long as it is extracted from a Drosophila mutant in which the expression of a specific gene is controlled by the UAS-GAL4 system, 1 is the T3dh (alcohol dehydrogenase type 3, type III alcohol dehydrogenase, CG3425) 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, T3dh RNAi activity was regulated to control T3dh gene expression and its function was confirmed.

The nucleotide sequence of SEQ ID NO: 1, the complementary nucleotide sequence of the nucleotide sequence of SEQ ID NO: 1, and the mRNA of any of the nucleotide sequences of SEQ ID NO: 1 and SEQ ID NO: 1 specifically promote senescence of Drosophila.

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, When the expression of the gene comprising at least one of the nucleotide sequences of SEQ ID NO: 1, the complementary nucleotide sequence thereof, and mRNA thereof is decreased by a factor of 2 or more, the aging is further confirmed , The above-mentioned gene can be used as a biomarker for discriminating the aging of a mammal or an insect other than a human or a human.

Furthermore, any one or more of the nucleotide sequences of SEQ ID NO: 1, their complementary nucleotide sequences, and mRNAs thereof may be used as the nucleotide sequence of a human ADHFE1 (alcohol dehydrogenase, iron containing 1, NP_653251.2 467 aa) gene The biomarker according to the present invention can be used to determine not only insects but also human progress of aging.

The nucleotide sequence of SEQ ID NO: 1 and the complementary nucleotide sequence of the nucleotide sequence of SEQ ID NO: 1 or the complementary nucleotide sequence of the nucleotide sequence of SEQ ID NO: 1 are complementary to each other. It is made of complementary 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 relates to a biomarker for obesity discrimination comprising at least one base sequence selected from the group consisting of the nucleotide sequence of SEQ ID NO: 1, the complementary base sequence thereof and the mRNA thereof.

The nucleotide sequence of SEQ ID NO: 1, the complementary nucleotide sequence thereof, and the mRNA of the nucleotide sequence of SEQ ID NO: 1 indicates that the diagnostic individual is obese or not (specifically, The increase in the size and density of the lipid sphere).

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, by providing at least one base sequence selected from the group consisting of the nucleotide sequence of SEQ ID NO: 1 in the present invention, the complementary base sequence thereof and the mRNA thereof as a biomarker for obesity discrimination, It is possible to accurately and simply determine the obesity of the diagnostic object.

The nucleotide sequence selected from the group consisting of SEQ ID NO: 1 is not particularly limited as long as it is extracted from a Drosophila mutant in which the expression of a specific gene is controlled by the UAS-GAL4 system, 1 is the T3dh (alcohol dehydrogenase type 3, type III alcohol dehydrogenase, CG3425) 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, T3dh RNAi activity was regulated to control T3dh gene expression and its function was confirmed.

The nucleotide sequence of SEQ ID NO: 1, the complementary base sequence thereof, and the mRNA thereof may be used as a result of the increase of the triglyceride content of the fruit fly and the size and density of lipids in fat tissue Expression decreases specifically.

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, It has been confirmed that obesity increases when the expression of the gene comprising any one or more base sequences selected from the group consisting of the nucleotide sequence of SEQ ID NO: 1, the complementary base sequence thereof and the mRNA thereof is reduced more than 2-fold The above-mentioned gene can be used as a biomarker for discriminating the obesity of a mammal or an insect other than a human or a human.

Furthermore, any one or more of the nucleotide sequences of SEQ ID NO: 1, their complementary nucleotide sequences, and mRNAs thereof may be used as the nucleotide sequence of a human ADHFE1 (alcohol dehydrogenase, iron containing 1, NP_653251.2 467 aa) gene It is possible to discriminate not only an insect but also a person's obesity using the biomarker according to the present invention.

The nucleotide sequence of SEQ ID NO: 1 and the complementary nucleotide sequence of the nucleotide sequence of SEQ ID NO: 1 or the complementary nucleotide sequence of the nucleotide sequence of SEQ ID NO: 1 are complementary to each other. It is made of complementary 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 relates to a biomarker for cancer diagnosis comprising at least one base sequence selected from the group consisting of the nucleotide sequence of SEQ ID NO: 1, the complementary base sequence thereof and the mRNA thereof.

A nucleotide sequence represented by SEQ ID NO: 1, a complementary base sequence thereof, and an mRNA thereof, can be used as a nucleotide sequence of a diagnostic individual for cancer development or proliferation (specifically, a phenotype of a cancer growth model Drosophila ), The expression level decreases.

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, by providing at least one base sequence selected from the group consisting of the nucleotide sequence of SEQ ID NO: 1 in the present invention, the complementary base sequence thereof and the mRNA thereof as the biomarker for cancer diagnosis, And it is possible to simply determine whether or not the diagnostic individual has cancer or growth.

The nucleotide sequence selected from the group consisting of SEQ ID NO: 1 is not particularly limited as long as it is extracted from a Drosophila mutant in which the expression of a specific gene is controlled by the UAS-GAL4 system, 1 is the T3dh (alcohol dehydrogenase type 3, type III alcohol dehydrogenase, CG3425) 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, T3dh RNAi activity was regulated to control T3dh gene expression and its function was confirmed.

The nucleotide sequence of SEQ ID NO: 1, the complementary base sequence thereof, and the mRNA thereof may be used as a result of the increase of the triglyceride content of the fruit fly and the size and density of lipids in fat tissue Expression decreases specifically.

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 And the expression of the gene comprising any one or more of the nucleotide sequences of SEQ ID NO: 1, their complementary base sequences and their mRNA sequences, It was confirmed that the cancer increased, and the above-mentioned gene can be used as a biomarker for diagnosing whether or not a mammal or an insect, other than human or human, has developed cancer.

Furthermore, any one or more of the nucleotide sequences of SEQ ID NO: 1, their complementary nucleotide sequences, and mRNAs thereof may be used as the nucleotide sequence of a human ADHFE1 (alcohol dehydrogenase, iron containing 1, NP_653251.2 467 aa) gene The biomarker according to the present invention can be used to diagnose not only insects but also human cancer or proliferation.

The nucleotide sequence of SEQ ID NO: 1 and the complementary nucleotide sequence of the nucleotide sequence of SEQ ID NO: 1 or the complementary nucleotide sequence of the nucleotide sequence of SEQ ID NO: 1 are complementary to each other. It is made of complementary 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, the biomarker comprising any one or more base sequences selected from the group consisting of the nucleotide sequence of SEQ ID NO: 1 according to the present invention, the complementary base sequence thereof and the mRNA thereof may be used for the determination of aging, Cancer diagnosis can be respectively detected, but at the same time, the progress of aging, the increase in obesity and the occurrence of cancer can be detected.

As described above, since the expression of one or more base sequences selected from the group consisting of the nucleotide sequence of SEQ ID NO: 1, the complementary base sequence thereof and the mRNA thereof is decreased, the UAS-GAl4 system is used to reduce specific gene expression Based on the progression of senescence of fruit fly mutants, the increase in lipid density and size in triglyceride and lipid body tissues, and the decrease in cancer incidence, the standardized biomarker generation And the amount of biomarker expressed in the diagnostic subject is compared to determine whether progress of aging, increase in obesity, and whether or not cancer has occurred.

Also, in the present invention, the biomarker may include not only the mRNA of the nucleotide sequence of SEQ ID NO: 1, the mRNA of the complementary nucleotide sequence of the same, And the protein may be any one or more selected from the group consisting of amino acid sequence 2. The term " amino acid sequence "

The protein consisting of the amino acid sequence 2 can also discriminate and diagnose any one selected from aging, obesity and cancer by increasing or decreasing the amount of protein compared with a normal control (standard expression amount of the same species as the diagnostic individual) .

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 a biomarker and a hybridization solution containing at least one base sequence selected from the group consisting of the nucleotide sequence of SEQ ID NO: 1, the complementary base sequence thereof and the mRNA thereof, As the expression of the biomarker decreases, the Drosophila mutant controlling the expression of a specific gene using the UAS-GAl4 system is further aged, the fat pore density and size in the triglyceride and fat body tissue are increased, It is possible to simultaneously detect the progress of aging, the increase in obesity, and the incidence of cancer by comparing the standard outbreak amount of the biomarker in the same species with the diagnostic biomarker expression amount in the diagnostic individual Which can be used as a complex diagnostic 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 NO: 1 or SEQ ID NO: 2 or an antibody capable of detecting the expression of any one of the amino acid sequences selected from the group consisting of SEQ ID NO: 2 Thereby detecting a protein comprising any one of the nucleotide sequences selected from the group consisting of SEQ ID NO: 1 or SEQ ID NO: 2, or a protein comprising any amino acid sequence selected from the group consisting of SEQ ID NO:

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 NO: 1 or SEQ ID NO: 2 or an antibody capable of detecting the expression of any one of the amino acid sequences selected from the group consisting of SEQ ID NO: 1 or a protein comprising any one of the amino acid sequences selected from the group consisting of SEQ ID NO: 2.

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 NO: 1 or SEQ ID NO: 2 or an antibody capable of detecting the expression of any one of the amino acid sequences selected from the group consisting of SEQ ID NO: 1 or a protein comprising any one of the amino acid sequences selected from the group consisting of SEQ ID NO: 2.

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 UAS-T3dh RNAi is transcribed by T3dh (Type III alcohol dehydrogenase, CG3425) when Gal4 is produced, resulting in decreased expression of T3dh due to RNAi action. T3dh mRNA levels were measured by RT-PCR in order to determine whether expression of T3dh was decreased in offspring (F1) fruit flies obtained by crossing Actin-GS-Gal4 with UAS-T3dh RNAi fruit flies. The amount of mRNA of T3dh Was significantly reduced compared to when it did not feed RU486. This result demonstrates that Actin-GS-Gal4 and UAS-T3dh RNAi are functioning normally (see FIG. 2 below).

[Example 1: Actin-GS-Gal4 > RT-PCR of UAS-T3dh RNAi]

Actin-GS-Gal4 Drosophila females and UAS-T3dh RNAi Drosophila males were crossed to determine whether the UAS-T3dh RNAi Drosophila was functioning normally. The males of F1 generation were collected and cultured in a medium containing RU486 (150 μM) , 10% Tegosept 1.6%, water 80.7%) and RU486 were grown for 10 days, and then the amount of T3dh mRNA was measured by reverse transcriptase polymerase Reverse transcriptase mediated PCR (RT-PCR) was performed. As a result, it was confirmed that the amount of T3dh mRNA in cultured fruit flies was significantly reduced compared to the control in the medium containing UAS-T3dh RNAi and containing RU486.

[Example 2: Actin-GS-Gal4 fed with RU486> Life test of UAS-T3dh RNAi Drosophila]

Actin-GS-Gal4 Drosophila females and UAS-T3dh 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-free medium. The lifespan was measured while growing at 25 ° C, 50% relative humidity, and 12:12 hour light cycle. This life test was repeated three times. As shown in the results of FIG. 3, it can be seen that as the expression of T3dh decreases, 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-T3dh RNAi Drosophila]

Actin-GS-Gal4 Drosophila females and UAS-T3dh RNAi Drosophila males were mated to collect males of F1 generation and grown for about 10 days on a medium containing RU486 (150 μM) and a medium containing no 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 triglyceride content of RU486-fed Drosophila, that is, the group in which T3dh expression was inhibited, was significantly increased (see FIG. 5 below).

[Example 4: Actin-GS-Gal4 fed with RU486] Analysis of liposome confocal microscope of UAS-T3dh RNAi fruit flies [

Actin-GS-Gal4 Drosophila females and UAS-T3dh RNAi Drosophila males were crossed to observe lipid size, and the males of F1 generation were collected and cultured for 10 days in media containing RU486 (150 μM) and 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: Production of cancer growth model flies (UAS-PI3K; c765-Gal4)

In order to carry out a study on cancer, c765-Gal4 Drosophila and UAS-PI3K Drosophila were crossed to produce cancer growth model flies.

[Example 6: phenotypic analysis of cancer growth model flies (UAS-PI3K; c765-Gal4)

Fabricated UAS-PI3K; C765-Gal4 / + CS10, UAS-PI3K / + CS10 and UAS-PI3K / + CS10 crossing wild type Drosophila (CS10) with c765-Gal4 and CS10 to analyze the phenotype of the c765-Gal4 cancer growth blackfly flies model. The wing length of c765-Gal4 / + CS10 was compared with UAS-PI3K / + 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 FIG. 8 below).

[Example 7: Influence of inhibition of T3dh gene expression on phenotype in cancer growth model floss (UAS-PI3K; c765-Gal4)]

In order to estimate the effect of T3dh gene expression inhibition on cancer growth, the wing phenotype of F1 generations obtained by crossing the cancer growth model flies (UAS-PI3K; c765-Gal4) with UAS-T3dh RNAi Drosophila was investigated. As a result, it was confirmed that the increased blade length and area due to PI3K overexpression decreased significantly when the expression of T3dh was suppressed (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> T3dh gene common associating life-reduction, cancer and obesity <130> HPC-5985 <160> 2 <170> KoPatentin 3.0 <210> 1 <211> 1395 <212> DNA <213> Drosophila sp. <400> 1 atgtcgcgga agaatgtctt aggtttgatc aacacgatcg tggccaactc ctgcaagtgc 60 cccgcccatt cgcataacta tggctcagca gctccaaccg cctcccaaac gggtcgcatg 120 gaatacgcct tcgagatgtc agcctcgact gttcgatttg gaccgggagt aagtgccgaa 180 gtgggagccg atcttcgcaa cttgggagct cggaaggttt gtttggtcac ggataaaaat 240 gtcgtgcaat tgccctccgt gaaagtggcc ttggattcac tggctcgtaa tggcatcaac 300 tacgaagtct atgatgaaac ccgggtggag cccacggatg ggagcatgtg gcatgccgtg 360 gagtttgcgc gcggcaagga gttcgatgca tttctggcca tcggaggagg atctgccatg 420 gatacagcca aggcagctaa tctttttagc agtgatgcaa atgcagagtt tttggattat 480 gtaaactgcc caattggcag aggcaaggag atatccgtaa aactgaagcc cctgattgcg 540 atgcccacca cttcgggaac aggttccgaa accactggag tagctatatt tgattacaag 600 aagttgcatg ccaaaactgg gatttccagc aagttcctca aacctacttt ggctgtgatt 660 gatcctctgc acaccctatc ccagccgcaa cgcgtgatgg ctttcgctgg ctttgatgtg 720 ttctgccatg ccctggagag tttcacggcg gtggattaca gggaacgcgg tttggcgccc 780 agtgatccca gcttgagacc cacctaccag ggcaggaacc cagtgtcgga tgtgtgggca 840 cgattcgcat tggagacgat aaggaaaaac tttgtgaacg ccatttacca gcccgataac 900 ctagaagccc gatcccagat gcacctggct tccacaatgg ctggcgtggg atttggtaat 960 gccggagtgc acctgtgtca tggcctttcc tatcctattt ctggaaatgt gagggattac 1020 ggatcacgct agtgctccgg cggtctttga gttcactgct ccagcttgtc cggatcggca tttggaggct 1140 gcccagttgc tgggcgcaga agtgcgaggt gtcgaaaaag cagacgccgg tcgtcttttg 1200 gcggacactg tacgcggttt tatgcaacgc gcgggcatag aaaatggcct ccgagagctg 1260 ggattctcca gcagcgatat acccgccttg gtggagggaa ccctgcccca ggaaaggatc 1320 actaagctag cacccagggc gcagacgcag gaaaacctgt cgcaactctt tgagaagtcc 1380 atggaggtct attaa 1395 <210> 2 <211> 464 <212> PRT <213> Drosophila sp. <400> 2 Met Ser Arg Lys Asn Val Leu Gly Leu Ile Asn Thr Ile Val Ala Asn   1 5 10 15 Ser Cys Lys Cys Pro Ala His Ser His Asn Tyr Gly Ser Ala Ala Pro              20 25 30 Thr Ala Ser Gln Thr Gly Arg Met Glu Tyr Ala Phe Glu Met Ser Ala          35 40 45 Ser Thr Val Arg Phe Gly Pro Gly Val Ser Ala Glu Val Gly Ala Asp      50 55 60 Leu Arg Asn Leu Gly Ala Arg Lys Val Cys Leu Val Thr Asp Lys Asn  65 70 75 80 Val Val Gln Leu Pro Ser Val Lys Val Ala Leu Asp Ser Leu Ala Arg                  85 90 95 Asn Gly Ile Asn Tyr Glu Val Tyr Asp Glu Thr Arg Val Glu Pro Thr             100 105 110 Asp Gly Ser Met Trp His Ala Val Glu Phe Ala Arg Gly Lys Glu Phe         115 120 125 Asp Ala Phe Leu Ala Ile Gly Gly Gly Ser Ala Met Asp Thr Ala Lys     130 135 140 Ala Ala Asn Ala Asp Ala Asp Ala Glu Phe Leu Asp Tyr 145 150 155 160 Val Asn Cys Pro Ile Gly Arg Gly Lys Glu Ile Ser Val Lys Leu Lys                 165 170 175 Pro Leu Ile Ala Met Pro Thr Thr Ser Gly Thr Gly Ser Glu Thr Thr             180 185 190 Gly Val Ala Ile Phe Asp Tyr Lys Lys Leu His Ala Lys Thr Gly Ile         195 200 205 Ser Ser Lys Phe Leu Lys Pro Thr Leu Ala Val Ile Asp Pro Leu His     210 215 220 Thr Leu Ser Gln Pro Gln Arg Val Met Ala Phe Ala Gly Phe Asp Val 225 230 235 240 Phe Cys His Ala Leu Glu Ser Phe Thr Ala Val Asp Tyr Arg Glu Arg                 245 250 255 Gly Leu Ala Pro Ser Asp Pro Ser Leu Arg Pro Thr Tyr Gln Gly Arg             260 265 270 Asn Pro Val Ser Asp Val Trp Ala Arg Phe Ala Leu Glu Thr Ile Arg         275 280 285 Lys Asn Phe Val Asn Ale Ile Tyr Gln Pro Asp Asn Leu Glu Ala Arg     290 295 300 Ser Gln Met His Leu Ala Ser Thr Met Ala Gly Val Gly Phe Gly Asn 305 310 315 320 Ala Gly Val His Leu Cys His Gly Leu Ser Tyr Pro Ile Ser Gly Asn                 325 330 335 Val Arg Asp Tyr Lys Pro Lys Gly Tyr Ser Ala Asp His Ala Leu Ile             340 345 350 Pro His Gly Leu Ser Val Val Ser Ser Ala Val A Phe Glu Phe         355 360 365 Thr Ala Pro Ala Cys Pro Asp Arg His Leu Glu Ala Ala Gln Leu Leu     370 375 380 Gly Ala Glu Val Gly Val Glu Lys Ala Asp Ala Gly Arg Leu Leu 385 390 395 400 Ala Asp Thr Val Arg Gly Phe Met Gln Arg Ala Gly Ile Glu Asn Gly                 405 410 415 Leu Arg Glu Leu Gly Phe Ser Ser Ser Asp Ile Pro Ala Leu Val Glu             420 425 430 Gly Thr Leu Pro Gln Glu Arg Ile Thr Lys Leu Ala Pro Arg Ala Gln         435 440 445 Thr Gln Glu Asn Leu Ser Gln Leu Phe Glu Lys Ser Met Glu Val Tyr     450 455 460

Claims (17)

A biomarker for discriminating the progress of aging comprising a nucleotide sequence represented by SEQ ID NO: 1, a complementary base sequence thereof, and a nucleotide sequence consisting of mRNA thereof. The method according to claim 1,
Wherein the amount of expression of the biomarker is decreased as the aging progress of the diagnostic subject is promoted. A biomarker for obesity discrimination comprising the nucleotide sequence represented by SEQ ID NO: 1, a complementary base sequence thereof, and any one of the nucleotide sequences consisting of mRNA thereof. 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. A cancer diagnostic biomarker comprising a nucleotide sequence represented by SEQ ID NO: 1, a complementary base sequence thereof, and any one of the nucleotide sequences consisting of mRNA thereof. 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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