KR101744397B1

KR101744397B1 - Use of Uba6 or Use1 as a diagnostic marker of cancer

Info

Publication number: KR101744397B1
Application number: KR1020150096321A
Authority: KR
Inventors: 이창환; 이태형; 김성진; 최세훈
Original assignee: 울산대학교 산학협력단
Priority date: 2015-07-07
Filing date: 2015-07-07
Publication date: 2017-06-08
Also published as: KR20170006016A

Abstract

The present invention relates to a composition for diagnosing cancer, and provides a cancer diagnostic composition excellent in sensitivity and specificity, which can be useful for early diagnosis of cancer by the present invention.

Description

Use of Uba6 or Use1 as a cancer diagnostic marker {Use of Uba6 or Use as a diagnostic marker of cancer}

The present invention relates to the use of Uba6 or Use1 as a cancer diagnostic marker.

Lung cancer is one of the most deadly cancers in the world, not only in Korea. This tendency is due to the absence of early diagnostic methods with low subjective symptoms and high sensitivity. Currently, lung cancer has a high dependence on imaging methods (X-ray, CT, MRI, etc.), and there are few examples of substances that can be used as biochemical indicators.

A biomarker is a term referring to all physical and biochemical indicators that can diagnose the physiological and pathological conditions of the body. Currently, the paradigm shifts from the paradigm of cancer treatment to induce early detection in the development of anticancer drugs that directly treat cancer, thereby enabling effective treatment and continuous monitoring. Therefore, what is most needed for such a change is a biomarker that can enable early diagnosis and monitoring of cancer.

Currently, there are substances listed for biomarkers for cancer diagnosis, but they still do not show sufficient specificity and sensitivity. Therefore, cross-reactivity exists in various diseases, and due to the lack of a method to complement them, the number of biomarkers practically applied to FDA approval in the US is very small.

Even in the case of lung cancer, a number of papers on biomarker candidates have been published and new biomarker candidate proteins have been studied, but so far, none have been commercialized as biomarkers capable of specifically diagnosing lung cancer.

U.S. Published Patent Application No. 2014-0323331 U.S. Published Patent Application No. 2014-0243211

SUMMARY OF THE INVENTION The present invention has been made to solve the above-mentioned problems, and it is an object of the present invention to quickly, easily and accurately diagnose cancer by identifying cancer-specific biomarkers.

Accordingly, it is an object of the present invention to provide a composition for diagnosing cancer, which comprises an agent for measuring the expression level of mRNA of Uba6 (Ubiquitin-like modifier activating enzyme 6) or Use1 (Uba6 specific E2) gene or its protein.

The present invention also provides a cancer diagnostic kit comprising the composition.

Further, the present invention relates to a method for detecting the expression level of Uba6 or Use1 gene, comprising the steps of: (a) measuring the expression level or the amount of protein of Uba6 or Use1 gene present in a biological sample; And (b) comparing the measurement result of step (a) with the expression level of the Uba6 or Use1 gene or the amount of the protein of the normal control sample, to provide information for prediction and diagnosis of cancer, There is a purpose.

In order to achieve the above object, the present invention provides a cancer diagnostic composition comprising an agent for measuring the expression level of mRNA of Uba6 (Ubiquitin-like modifier activating enzyme 6) or Use1 (Uba6 specific E2) gene or its protein do.

In one embodiment of the present invention, the agent for measuring the expression level of the gene mRNA may include a primer that specifically binds to Uba6 or Use1 gene.

In one embodiment of the present invention, the agent for measuring the expression level of the gene mRNA may include a probe specifically binding to the Uba6 or Use1 gene.

In one embodiment of the present invention, the agent for measuring the level of expression of the protein may comprise an antibody specific for Uba6 or Use1 protein.

In one embodiment of the present invention, the kit may be an RT-PCR kit, a DNA chip kit, or a protein chip kit.

(A) measuring an expression level or amount of protein of Uba6 or Use1 gene present in a biological sample; And (b) comparing the measurement result of step (a) with the expression level of the Uba6 or Use1 gene of the normal control sample or the amount of the protein, to provide information for prediction and diagnosis of cancer.

In one embodiment of the present invention, the biological sample is selected from the group consisting of tissue, cells, blood, serum, plasma, saliva, and urine.

In one embodiment of the present invention, the measurement is performed using a reverse transcriptase-polymerase chain reaction, a real time-polymerase chain reaction, a Western blot, a Northern blot, an enzyme- linked immunosorbent assay, radioimmunoassay (RIA), radioimmunoprecipitation and immunoprecipitation assays.

In one embodiment of the present invention, when the amount of the Uba6 or Use1 gene expressed in the biological sample or the amount of the protein is increased as compared with the control, it may be determined that the cancer has occurred.

The present invention provides a cancer diagnostic composition excellent in sensitivity and specificity that can be useful for early diagnosis of cancer.

FIG. 1 shows the result of examining the expression pattern of Uba6 and Use1 gene in 105 pairs of cancerous and normal tissues.
FIG. 2 shows the results of statistical analysis of the expression patterns of Uba6 and Use1 gene in 105 pairs of cancerous and normal tissues.
FIG. 3 is a result of confirming the expression pattern of TC1 cells after overexpressing Uba6 or Use1.
FIG. 4 shows the results of confirming the degree of cell aggregation overexpressing Uba6, Use1.
FIG. 5 shows the results of confirming the degree of proliferation of cells overexpressing Uba6, Use1.
FIG. 6 shows the result of confirming the wound healing level of cells overexpressing Uba6, Use1.
FIG. 7 shows the results of confirming the degree of cell transduction in which Uba6, Use1 was overexpressed.
FIG. 8 shows the results of confirming the degree of cancer formation after xenograft transplantation into cells overexpressing Uba6, Use1.
FIG. 9 shows the results of analysis of overexpression of Uba6 or Use1 in lung cancer tissues of lung cancer patients.

The present invention confirms whether abnormal expression of Uba6 or Use1 is associated with cancer development in cancer tissues and confirms that expression of Uba6 or Use1 gene is overexpressed in cancer tissues and cancer cells compared with normal tissues and normal cells, It is characterized by the fact that the gene can be used as a biomarker for cancer diagnosis.

In other words, the present inventors confirmed the expression patterns of UBA6 and USE1 genes in 105 pairs of cancer and normal tissues and statistically analyzed the expression patterns of UBA6 and USE1, and found that UBA6 and USE1 were overexpressed by 83% and 92%, respectively, in cancer patients (See FIG. 2).

According to one embodiment of the present invention, it was confirmed that the UBA6 and USE1 genes rapidly formed cell bodies faster than overexpressed TC1 cells (see FIG. 4).

According to another embodiment of the present invention, it was found that the TC1 cells overexpressing UBA6 and USE1 genes proliferate more rapidly than the cells not overexpressed (see FIG. 5).

According to another embodiment of the present invention, the wound healing ability is superior to that of cells in which TC1 cells overexpressing UBA6 and USE1 genes, respectively, are not overexpressed (see FIG. 6).

According to another embodiment of the present invention, it was confirmed that the cells were able to form cancer cells more than TC1 cells overexpressing UBA6 and USE1 genes, respectively (see FIG. 7).

As a result, the present inventors have found that UBA6 and USE1 genes are overexpressed in cancers, and that artificially overexpressing each gene in cells leads to an increase in proliferation, mobility and invasiveness, which are characteristic of cancer. In addition, xenotransplantation of the cells overexpressing the UBA6 and USE1 genes into nude mice confirmed that the mice developed cancer.

Therefore, the UBA6 or USE1 gene of the present invention can be used as a composition for cancer diagnosis. Therefore, the present invention provides a method for detecting the expression level of mRNA of Uba6 (Ub6-like modifier activating enzyme 6) or Use1 (Uba6 specific E2) A composition for diagnosing cancer comprising the agent to be measured can be provided.

In the present invention, the UBA6 gene may have the nucleotide sequence shown in SEQ ID NO: 1, and the UBA6 protein may have the amino acid sequence shown in SEQ ID NO: 2. Also, in the present invention, the USE1 gene may have the nucleotide sequence shown in SEQ ID NO: 3, and the USE1 protein may have the amino acid sequence shown in SEQ ID NO: 4.

In the present invention, the term " diagnosis " means confirming a pathological condition. For the purpose of the present invention, the diagnosis means checking whether cancer marker is present or not, , The diagnosis of the present invention includes determining whether or not the cancer is onset, development and alleviation by confirming the presence or absence and expression level of the cancer diagnosis marker.

The term "cancer" used in the present invention includes gastric cancer, colon cancer, breast cancer, lung cancer, non-small cell lung cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, skin or intraocular melanoma, uterine cancer, ovarian cancer, Endometrioid cancer, thyroid cancer, pituitary cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, uterine cancer, endometrial carcinoma, endometrial carcinoma, endometrial carcinoma, endometrial cancer, uterine cancer, vulvar carcinoma, vulvar carcinoma, , At least one disease selected from the group consisting of chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, renal or urothelial cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma and pituitary adenoma But are not limited to, lung cancer.

The diagnostic marker means a substance capable of distinguishing cancer cells from normal cells and includes a polypeptide or nucleic acid (for example, mRNA or the like) which shows an increase or decrease in cancer cells as compared with normal cells ), Lipids, glycolipids, glycoproteins, sugar (monosaccharides, disaccharides, oligosaccharides, etc.) and the like. The marker for cancer diagnosis provided by the present invention may be a Uba6 or Use1 gene or protein whose expression level is increased in cancer cells as compared with normal cells.

In the present invention, the level of the Uba6 or Use1 gene preferably indicates the level of mRNA in which the Uba6 or Use1 gene is expressed, that is, the amount of mRNA. As the substance capable of measuring the level, Uba6 or Use1 gene- Primers or probes. In the present invention, the primer or the probe specific to the Uba6 gene may be a primer or a probe capable of specifically amplifying the entire Uba6 gene or a specific region of the gene of SEQ ID NO: 1, You can design through methods.

In addition, in the present invention, the primer or probe specific for Use1 gene may be a primer or a probe capable of specifically amplifying the entire Use1 gene or a specific region of the gene shown in SEQ ID NO: 3, Can be designed through methods known in the art.

In the present invention, the term 'primer' refers to a primer that can function as a starting point for template-directed DNA synthesis under suitable conditions (ie, four different nucleoside triphosphates and polymerization reaction enzymes) - means strand oligonucleotide. The appropriate length of the primer may vary depending on various factors, such as temperature and use of the primer. In addition, the sequence of the primer does not need to have a sequence completely complementary to a partial sequence of the template, and it is sufficient that the primer has sufficient complementarity within a range capable of hybridizing with the template and acting as a primer. Therefore, the primer in the present invention does not need to have a perfectly complementary sequence to the nucleotide sequence of the Uba6 or Use1 gene as a template, and it is sufficient that the primer has sufficient complementarity within a range capable of hybridizing with the gene sequence and acting as a primer. In addition, it is preferable that the primer according to the present invention can be used for gene amplification reaction. The amplification reaction refers to a reaction for amplifying a nucleic acid molecule. The amplification reactions of these genes are well known in the art, and examples thereof include polymerase chain reaction (PCR), reverse transcription polymerase chain reaction (RT-PCR) (LCR), electron mediated amplification (TMA), nucleic acid sequencing substrate amplification (NASBA), and the like.

In the present invention, the term " probe " refers to a linear oligomer of natural or modified monomers or linkages, including deoxyribonucleotides and ribonucleotides, capable of specifically hybridizing to a target nucleotide sequence, Or artificially synthesized. The probes according to the present invention may be single-stranded, preferably oligodeoxyribonucleotides. Probes of the invention can include natural dNMPs (i.e., dAMP, dGMP, dCMP and dTMP), nucleotide analogs or derivatives. In addition, the probe of the present invention may also include a ribonucleotide. For example, the probes of the present invention can be used in combination with a framework-modified nucleotide such as a peptide nucleic acid (PNA) (M. Egholm et al., Nature, 365: 566-568 (1993)), phosphorothioate DNA, phosphorodithioate DNA, phosphoamidate DNA, amide-linked DNA, MMI-linked DNA, 2'-O-methyl RNA, alpha-DNA and methylphosphonate DNA, sugar modified nucleotides such as 2'- 2'-O-alkyl DNA, 2'-O-allyl DNA, 2'-O-alkynyl DNA, hexose DNA, pyranosyl RNA, and anhydrohexy Tolyl DNA, and nucleotides with base modifications such as C-5 substituted pyrimidines wherein the substituents are fluoro, bromo, chloro, iodo-, methyl-, ethyl-, vinyl-, formyl-, 7-deazapurines having C-7 substituents (the substituents being fluoro, bromo, chloro, bromo, chloro, , Iodo-, me -, ethyl-, vinyl-, formyl-, alkynyl -, alkenyl-, quinolyl and quinoxalyl thiazol-, quinolyl and quinoxalyl imidazolidin -, pyridyl-), inosine, and may include a diamino purine.

Also, in the present invention, the substance capable of measuring the level of the Uba6 or Use1 protein may include an antibody such as a polyclonal antibody, a monoclonal antibody and a recombinant antibody capable of specifically binding to Uba6 or Use1 protein have.

In the present invention, since Uba6 or Use1 protein has been identified as a marker protein capable of diagnosing cancer as described above, a method for producing an antibody using the protein can be performed by a technique known to those skilled in the art Can be easily manufactured by using the above method. For example, in the case of polyclonal antibodies, Uba6 or Use1 antigens can be produced by methods well known in the art for obtaining sera containing antibodies by injection into an animal and blood sampling from the animal. Such polyclonal antibodies include chlorine, rabbit , Sheep, monkeys, horses, pigs, cows, dogs, and the like. Monoclonal antibodies can be prepared using the hybridoma method (Kohler et al., European Jounal of Immunology, 6, 511-519, 1976) well known in the art or can be prepared using the phage antibody library Clackson et al., Nature, 352, 624-628, 1991, Marks et al., J. Mol. Biol., 222: 58, 1-597, 1991). In addition, an antibody according to the present invention may comprise a functional fragment of an antibody molecule as well as a complete form having two full-length light chains and two full-length heavy chains. A functional fragment of an antibody molecule refers to a fragment having at least an antigen binding function, and includes Fab, F (ab ') 2, F (ab') 2 and Fv.

The present invention also provides a cancer diagnostic kit comprising a cancer diagnostic composition capable of measuring the expression level of Uba6 or Use1 protein or a gene encoding the same.

The cancer diagnostic composition contained in the cancer diagnostic kit of the present invention may include a primer, a probe or an antibody capable of measuring the expression level of the Uba6 or Use1 protein or a gene encoding the Uba6 or Use1 protein, .

When the cancer diagnostic kit of the present invention is applied to a PCR amplification process, the kit of the present invention may optionally include a reagent necessary for PCR amplification such as a buffer, a DNA polymerase (e.g., Thermus aquaticus (Taq), Thermus thermophilus ), Thermostable DNA polymerase obtained from Thermus filiformis, Thermis flavus, Thermococcus literalis or Pyrococcus furiosus (Pfu), DNA polymerase joins and dNTPs, and the cancer diagnostic kit of the present invention is applied to immunoassay In this case, the kit of the present invention may optionally comprise a secondary antibody and a labeling substrate.

Further, the kit according to the present invention may be manufactured from a number of separate packaging or compartments containing the reagent components described above.

The present invention also provides a cancer diagnostic microarray comprising a cancer diagnostic composition capable of measuring the expression level of Uba6 or Use1 protein or a gene encoding the same.

In the microarray of the present invention, a primer, a probe or an antibody capable of measuring the expression level of the Uba6 or Use1 protein or a gene encoding the Uba6 or Use1 protein is used as a hybridizable array element and immobilized on a substrate do. Preferred substrates may include, for example, membranes, filters, chips, slides, wafers, fibers, magnetic beads or nonmagnetic beads, gels, tubing, plates, polymers, microparticles and capillaries, as suitable rigid or semi-rigid supports have. The hybridization array elements are arranged and immobilized on the substrate, and such immobilization can be performed by chemical bonding methods or covalent bonding methods such as UV. For example, the hybridization array element may be bonded to a glass surface modified to include an epoxy compound or an aldehyde group, and may also be bound by UV on a polylysine coating surface. In addition, the hybridization array element can be coupled to the substrate via a linker (e.g., ethylene glycol oligomer and diamine).

On the other hand, when the sample to be applied to the microarray of the present invention is a nucleic acid, it may be labeled and hybridized with an array element on a microarray.

Hybridization conditions may vary, and detection and analysis of hybridization degree may be variously performed depending on the labeled substance.

The present invention also provides a method of measuring the expression level of Uba6 or Use1 gene or the level of Uba6 or Use1 protein to provide information for predicting and diagnosing cancer, said method comprising the steps of: (a) Measuring the expression level or the amount of protein of Uba6 or Use1 gene; And (b) comparing the measurement result of step (a) with the expression level or the amount of protein of the Uba6 or Use1 gene of the normal control sample. The method of measuring the expression level of the Uba6 or Use1 gene or the Uba6 or Use1 protein level in the above can be carried out by a known process for separating mRNA or protein from a biological sample using a known technique.

In the present invention, the "biological sample" refers to a sample collected from a living body different from the normal control group in the expression level or protein level of the Uba6 or Use1 gene according to the degree of cancer development or progression. Examples of the sample include But are not limited to, tissues, cells, blood, serum, plasma, saliva and urine.

The expression level of the Uba6 or Use1 gene is preferably determined by measuring the level of mRNA. The level of the mRNA may be measured by RT-PCR, RT-PCR, RNase protection Methods, Northern blots and DNA chips, but are not limited thereto.

The Uba6 or Use1 protein level can be measured by using an antibody. In this case, the Uba6 or Use1 marker protein in the biological sample and the antibody specific thereto form a conjugate, that is, an antigen-antibody complex and the antigen- The amount of formation can be measured quantitatively through the size of the signal of the detection label. Such detection labels can be selected from the group consisting of enzymes, minerals, ligands, emitters, microparticles, redox molecules and radioisotopes, but is not limited thereto.

Analytical methods for measuring protein levels include, but are not limited to, Western blotting, ELISA, radioimmunoassay, radioimmunoassay, Ouchteroni immunodiffusion, rocket immunoelectrophoresis, tissue immuno staining, immunoprecipitation assays, Complement fixation assay, FACS, and protein chip.

Therefore, the present invention can confirm the amount of Uba6 or Use1 mRNA expression or the amount of protein in the control group and the amount of Uba6 or Use1 mRNA expression or protein in the cancer patient or suspected cancer patient through the detection methods as described above, And the degree of the cancer is compared with the control group, it is possible to predict and diagnose the onset, progress stage or prognosis of the cancer.

Hereinafter, the present invention will be described in detail with reference to examples. However, these examples are intended to further illustrate the present invention, and the scope of the present invention is not limited to these examples.

< Example 1>

In lung cancer tissue UBA6 Wow USE1 The expression pattern of

For this experiment, 105 pairs of lung-squamous cell carcinoma (lung-adenocarcinoma) patients were received on November 3, 2014 and December 24, 2014 at the Asan Medical Center's tissue resource center (IRB: 2014- 0960).

FIG. 1 shows the expression patterns of UBA6 and USE1 genes in 105 lung cancer and normal tissues. As a result of statistical analysis of the expression patterns of these genes, UBA6 and USE1 were overexpressed in lung cancer patients 83% and 92%, respectively (See FIG. 2).

< Example 2>

Western Blot Experiment

The tissue prepared by the method of Example 1 was dissolved in 200 μl of a lysis buffer supplemented with a protease inhibitor cocktail (Complete Mini EDTA-free; Roche) in NP40 lysis buffer (Tris 50 mM, NaCl 150 mM, NP40 0.5% The mixture was homogenized and incubated at 4 ° C for 30 minutes.

The supernatant was transferred to a new microcentrifuge tube by centrifugation at 13,000 x g for 30 min and the protein concentration was measured with the BCA protein assay kit (Thermo). 30 μg of protein was mixed with 5X loading buffer, heated at 95 ° C for 5 minutes, and then subjected to 8% SDS-PAGE (sodium dodecylsulfate-poly acrylamide gel electrophoresis).

After electrophoresis, the gel protein was transferred to a polyvinylidenedifluoride transfer membrane (Immun-blot PVDF membrane for protein blotting; Bio RAD) at 100 mV for 1 hour. The membranes were blocked with PBST (0.01% Tween 20 in PBS) in 5% skim milk solution for 1 hour and then washed 3 times every 10 minutes with PBST. Then, the primary antibody was diluted in 5% skim milk, subjected to O / N reaction at 4 ° C, and then washed three times with PBST every 10 minutes. Secondary antibody conjugated with horseradish peroxidase was diluted in 5% skim milk, reacted at room temperature for 1 hour, and washed 3 times with PBST every 10 minutes. In addition, the membrane was exposed to an X-ray film using an ECL (clarity western ECL substrate; BIO-RAD) to confirm the expression pattern of a specific protein (see FIG. 3).

< Example 3>

Stable cell construction

TC1 cells were cultured in DMEM / High Glucose (HyClone, SH30243.01) supplemented with 10% Fetal Bovine Serum (hyClone, SH30919.03) and 1% penicillin-streptomycin (HyClone, SV30010). The retroviral system was used to overexpress the UBA6 and USE1 genes in TC1 cells.

UCA6 and USE1 primers containing specific sequences were PCR (polymerase chain reaction) using KOD Hot start DNA Polymerase (Novagen, 71086). The PCR products were then used to generate UBA6 and USE1 products, and the products were fused to the pDONR223 vector through the BP reaction, a gateway system. After that, E. coli DH5α was transformed and then stained on Lunia-Bertani (LB) agar plate with Spectinomycin added and cultured at 37 ° C. The plasmids were purified using DNA-Spin (iNtRON, 17087) after growing colonies on LB broth.

UBA6 and USE1 genes were identified as BsrG I (BioLabs, R0575L), one of the restriction enzymes. The confirmed UBA6 and USE1 genes were fused to the MSCV-N-HA-Flag Puro vector through the LR reaction, which is a gateway system, to make stable cells using retroviruses. The presence of the gene was confirmed once again with BsrGl. The retroviruses were transfected with 293T cells using an iNfect transfection reagent (iNtRON, 15081) after mixing 1: 1: 1 of the MSCV vector and its helper DNA, Gag / Pol, VSVG. The next day, the medium was replaced with a new medium. On the second day, TC1 cells were infected with polyblene and then replaced with fresh medium the next day. On the third day after the injection, puromycin (R & D, 4089) was added to the medium at 2 ug / ml, and the medium was changed every 2-3 days. In the same manner, UBA6 and USE1 were constructed in A549 cells.

< Example 4>

UBA6 , USE1 &Lt; / RTI > stock Formation analysis

TC1 cells were washed once with PBS solution and then trypsinized for 1 minute at 37 ° C in a CO ₂ incubator. The removed TC1 cells were centrifuged at 1,200 rpm for 5 minutes. The cells were mixed using a culture solution, and 1,000 cells were prepared per ml using a hemacytometer. 1 ml of TC1 cells prepared in a 6-well plate to which 2 ml of the culture had been added in advance was added and mixed well. The cells were cultured in a 37 ° C CO ₂ incubator for 10 to 20 days. The culture medium was removed, washed once with PBS, and fixed with 3.7% PFA (EMS, 15710-S) for 5 minutes. After staining with 0.05% crystal violet (Sigma-Aldrich, C3886) solution for 30 minutes, the plate dye was removed using tap water, and the plate was allowed to dry until it was scanned or photographed.

As a result of confirming the degree of cell formation of cells overexpressing UBA6 and USE1 as described above, it was confirmed that UBA6 and USE1 form cells more rapidly than control cells not overexpressing TC1 cells (see FIG. 4).

< Example 5>

UBA6 , USE1 Proliferation of cells overexpressed

The proliferation ability of TC1 cells was evaluated using CellTiter-Glo Luminescenc Cell Viability Assay (Promega). TC1 cells were washed with PBS and then trypsinized for 1 minute at 37 ° C in a CO ₂ incubator.

The removed TC1 cells were centrifuged at 1,200 rpm for 5 minutes. Cells were mixed using a culture solution, and 10,000 cells per 100 μl were prepared using a hemacytometer. 100 μl of cell suspension per cell was added to the 96 well black or white solid plate (Costar, 3917) three times in wells. Cells were cultured in a CO ₂ incubator at 37 ° C, and the plate was stabilized at room temperature for 30 minutes for analysis. 100 CellTiter-Glo reagent (Promega, G755A) was added to each well and the cells were lysed using a shaker for 2 minutes. The reaction was carried out at room temperature for 10 minutes and then the value was obtained using a GloMax 96 Microplate Luminometer (Promega).

As a result of confirming the degree of cell proliferation of UBA6 and USE1 overexpressed as described above, it was found that UBA6 and USE1 proliferate more rapidly than control cells overexpressing TC1 cells (see FIG. 5).

< Example 6>

UBA6 , USE1 Of wound healing ability of overexpressed cells

Wound healing assay was performed to examine the migration ability of TC1 cells. TC1 cells were washed once with PBS solution and then trypsinized in a CO ₂ incubator at 37 ° C for 1 min. The removed TC1 cells were centrifuged at 1,200 rpm for 5 minutes, and the cells were again mixed using the culture solution, and then 500,000 cells were prepared per ml using a hemacytometer.

After one or two lines were drawn on the bottom of the 6-well plate, 1 ml of the culture solution was added to each well. After addition of 1 ml of prepared TC1 cells, the plate was shaken evenly, and cells were then cultured in a CO ₂ incubator at 37 ° C. When the cell density was 80 to 90%, the cells were carefully drawn using a 200P tip and washed carefully twice with the culture medium. Then, a photograph of 100 times of 0 hours was obtained using a microscope (NiKon eclipse Ti-U). To obtain the following photographs, the cells were washed once with the warmed culture medium, and then photographs were taken. Analysis of the photographs was performed using an Image J program.

As a result of confirming the wound healing level of cells overexpressing UBA6 and USE1 as described above, it was found that the wound healing ability was superior to the control cells in which TC1 cells overexpressing UBA6 and USE1 were not overexpressed (FIG. 6 Reference).

< Example 7>

UBA6 , USE1 &Lt; RTI ID = 0.0 > transient < / RTI >

Transwell invasion assay was performed to determine the invasive ability of TC1 cells as cancer.

Matrigel Matrix (Corning, C354234) was diluted to a concentration of 2 ug / ml in 24-well cuture inserts (SPL, 35224) and then 120 ul of Matrigel was entirely coated on the membrane in the insert. Gelled in a CO ₂ incubator. On the next day, a culture medium supplemented with 800 μl of FBS was added to the inner well of the insert. And placed in a CO ₂ incubator at 37 ° C for 3 hours. TC1 cells were washed once with PBS solution and then trypsinized in a CO ₂ incubator at 37 ° C for 1 min. The removed TC1 cells were centrifuged at 1,200 rpm for 5 minutes, and the cells were again mixed using the culture solution, and 100,000 cells were prepared per 500 μl using a hemacytometer. At this time, the culture solution was mixed with serum-free (serum-free culture). The cells were incubated in a CO ₂ incubator at 37 ° C for 36 hours.

After 36 hours, the insert was washed twice with PBS and fixed with 3.7% PFA / 5.4% sucrose for 15 minutes. After washing twice with PBS, place in methanol for 20 minutes. After 20 minutes, the cells were washed twice with PBS, and stained with 0.5% crystal violet solution for 10 minutes. After washing twice with PBS, non-invaded cells in the insert were carefully removed using a cotton swab. The inserts were dried for about one hour, then placed on a slide and observed with a microscope.

As a result, UBA6 and USE1 overexpressed cells were transfected more rapidly than control cells overexpressing TC1 cells (see FIG. 7).

< Example 8>

carcinoma Xenotransplantation model

A549 cells overexpressing Uba6, Use1 and control were cultured and then concentrated to 5 x 10 ⁶ cells / 200 μl in PBS. Six-week-old Balb / c nude mice were subcutaneously injected with 200 μl of cells in the right side of the waist, and calves were calibrated for 30 days every 3 days using a caliper.

As a result, it was found that cancer was formed in cells that were not overexpressed in A549 cells in which Uba6 or Use1 was overexpressed, respectively (see FIG. 8).

As a result, it was confirmed that the Uba6 or Use1 gene of the present invention can be used for cancer diagnosis including lung cancer.

< Example 9>

UBA6 And USE1 Of lung cancer Diagnostic marker Proof of availability

Furthermore, the present inventors conducted the following experiment to confirm that Uba6 and Use1 can be practically used as biomarkers for diagnosis of lung cancer. A total of 105 patients diagnosed with lung cancer at Asan Medical Center were analyzed by Western blot analysis for the expression of Uba6 and Use1 in tissues after obtaining lesion lung tissue and normal lung tissue from these patients.

As a result, the expression of Uba6 or Use1 was increased in 102 patients except for 3 of 105 patients with lung cancer, and Uba6 and Use1 were overexpressed in 82 patients.

Therefore, the inventors of the present invention have found that the expression level of Uba6 or Use1 can be used to confirm the onset of lung cancer. Further, when the diagnosis is made using Uba6 and Use1 together, the accuracy and probability of diagnosis can be increased (See FIG. 9).

The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

<110> Industry Academic Cooperation Foundation of Ulsan University <120> Use of Uba6 or Use 1 as a diagnostic marker of cancer <130> pn1505-130 <160> 4 <170> Kopatentin 2.0 <210> 1 <211> 3159 <212> DNA &Lt; 213 > UBA6 cDNA sequence <400> 1 atggaaggat ccgagcctgt ggccgcccat cagggggaag aggcgtcctg ttcttcctgg 60 gggactggca gcacaaataa aaatttgccc attatgtcaa cagcatctgt ggaaatcgat 120 gatgcattgt atagtcgaca gaggtacgtt cttggagaca cagcaatgca gaagatggcc 180 aagtcccatg ttttcttaag tgggatgggt ggtcttggtt tggaaattgc aaagaatctt 240 gttcttgcag ggattaaggc agttacaatt catgatacag aaaaatgcca agcatgggat 300 ctaggaacca acttctttct cagtgaagat gatgttgtta ataagagaaa cagggctgaa 360 gctgtactta aacatattgc agaactaaat ccatacgttc atgtcacatc atcttctgtt 420 cctttcaatg agaccacaga tctctccttt ttagataaat accagtgtgt agtattgact 480 gagatgaaac ttccattgca gaagaagatc aatgactttt gccgttctca gtgccctcca 540 attaagttta tcagtgcaga tgtacatgga atttggtcaa ggttattttg tgatttcggt 600 gatgaatttg aagttttaga tacaacagga gaagaaccaa aagaaatttt catttcaaac 660 ataacgcaag caaatcctgg cattgttact tgccttgaaa atcatcctca caaactggag 720 acaggacaat tcctaacatt tcgagaaatt aatggaatga caggtttaaa tggatctata 780 caacaaataa cggtgatatc gccattttct tttagtattg gtgacaccac agaactggaa 840 ccatatttac atggaggcat agctgtccaa gttaagactc ctaaaacagt tttttttgaa 900 tcactggaga ggcagttaaa acatccaaag tgccttattg tggattttag caaccctgag 960 gcacctttag agattcacac agctatgctt gccttggacc agtttcagga gaaatacagt 1020 cgcaagccaa atgttggatg ccaacaagat tcagaagaac tgttgaaact agcaacatct 1080 ataagtgaaa ccttggaaga gaagcctgat gtaaatgctg acattgtgca ttggctctct 1140 tggactgccc aaggcttttt atctccactt gctgcagcag taggaggtgt tgccagccaa 1200 gaagtattga aagctgtaac aggaaaattt tctcctttgt gccagtggtt atatcttgaa 1260 gcagcagata ttgttgaatc actaggcaaa cctgaatgtg aagaatttct cccacgagga 1320 gatagatatg atgccttaag agcttgcatt ggagacactt tgtgtcagaa actgcaaaat 1380 ttaaacatct tcttagtagg gtgtggagcc ataggctgtg aaatgttgaa aaattttgct 1440 ttactggtg ttggcacaag caaagagaaa ggaatgatta cagttacaga tcctgacttg 1500 atagagaaat ccaacttaaa tagacagttc ctatttcgtc ctcatcacat acagaaacct 1560 aaaagctaca ctgctgctga tgctactctg aaaataaatt ctcaaataaa gatagatgca 1620 cacctgaaca aagtatgtcc aaccactgag accatttaca atgatgagtt ctatactaaa 1680 caagatgtaa ttattacagc attagataat gtggaagcca ggagatacgt agacagtcgt 1740 tgcttagcaa atctaaggcc tcttttagat tctggaacaa tgggcactaa gggacacact 1800 gaagttattg taccgcattt gactgagtct tacaatagtc atcgggatcc cccagaagag 1860 gaaataccat tttgtactct aaaatccttt ccagctgcta ttgaacatac catacagtgg 1920 gcaagagata agtttgaaag ttccttttcc cacaaacctt cattgtttaa caaattttgg 1980 caaacctatt catctgcaga agaagtctta cagaagatac agagtggaca cagtttagaa 2040 ggctgttttc aagttataaa gttacttagc agaagaccta gaaattggtc ccagtgtgta 2100 gaattagcaa gattaaagtt tgaaaaatat tttaaccata aggctcttca gcttcttcac 2160 tgtttccctc tggacatacg attaaaagat ggcagtttat tttggcagtc accaaagagg 2220 ccaccctctc caataaaatt tgatttaaat gagcctttgc acctcagttt ccttcagaat 2280 gctgcaaaac tatatgctac agtatattgt attccatttg cagaagagga cttatcagca 2340 gatgccctct tgaatattct ttcagaagta aagattcagg aattcaagcc ttccaataag 2400 gttgttcaaa cagatgaaac tgcaaggaaa ccagaccatg ttcctattag cagtgaagat 2460 gagaggaatg caattttcca actagaaaag gctattttat ctaatgaagc caccaaaagt 2520 gaccttcaga tggcagtgct ttcatttgaa aaagatgatg atcataatgg acacatagat 2580 ttcatcacag ctgcatcaaa tcttcgtgcc aaaatgtaca gcattgaacc agctgaccgt 2640 ttcaaaacaa agcgcatagc tggtaaaatt atacctgcta tagcaacaac cactgctaca 2700 gtttctggct tggttgcctt ggagatgatc aaagtaactg gtggctatcc atttgaagct 2760 tacaaaaatt gttttcttaa cttagccatt ccaattgtag tatttacaga gacaactgaa 2820 gtaaggaaaa ctaaaatcag aaatggaata tcatttacaa tttgggatcg atggaccgta 2880 catggaaaag aagatttcac cctcttggat ttcataaatg cagtcaaaga gaagtatgga 2940 attgagccaa caatggtggt acagggagtc aaaatgcttt atgttcctgt aatgcctggt 3000 catgcaaaaa gattgaagtt aacaatgcat aaacttgtaa aacctactac tgaaaagaaa 3060 tatgtggatc ttactgtgtc atttgctcca gacattgatg gagatgaaga tttgccggga 3120 cctccagtaa gatactactt cagtcatgac actgattaa 3159 <210> 2 <211> 1052 <212> PRT <213> UBA6 Protein sequence <400> 2 Met Glu Gly Ser Glu Pro Val Ala Ala His Gln Gly Glu Glu Ala Ser 1 5 10 15 Cys Ser Ser Trp Gly Thr Gly Ser Thr Asn Lys Asn Leu Pro Ile Met 20 25 30 Ser Thr Ala Ser Val Glu Ile Asp Asp Ala Leu Tyr Ser Arg Gln Arg 35 40 45 Tyr Val Leu Gly Asp Thr Ala Met Gln Lys Met Ala Lys Ser His Val 50 55 60 Phe Leu Ser Gly Met Gly Gly Leu Gly Leu Glu Ile Ala Lys Asn Leu 65 70 75 80 Val Leu Ala Gly Ile Lys Ala Val Thr Ile His Asp Thr Glu Lys Cys 85 90 95 Gln Ala Trp Asp Leu Gly Thr Asn Phe Phe Leu Ser Glu Asp Asp Val 100 105 110 Val Asn Lys Arg Asn Arg Ala Glu Ala Val Leu Lys His Ile Ala Glu 115 120 125 Leu Asn Pro Tyr Val His Val Thr Ser Ser Ser Val Pro Phe Asn Glu 130 135 140 Thr Thr Asp Leu Ser Phe Leu Asp Lys Tyr Gln Cys Val Val Leu Thr 145 150 155 160 Glu Met Lys Leu Pro Leu Gln Lys Lys Ile Asn Asp Phe Cys Arg Ser 165 170 175 Gln Cys Pro Pro Ile Lys Phe Ile Ser Ala Asp Val His Gly Ile Trp 180 185 190 Ser Arg Leu Phe Cys Asp Phe Gly Asp Glu Phe Glu Val Leu Asp Thr 195 200 205 Thr Gly Glu Glu Pro Lys Glu Ile Phe Ile Ser Asn Ile Thr Gln Ala 210 215 220 Asn Pro Gly Ile Val Thr Cys Leu Glu Asn His Pro His Lys Leu Glu 225 230 235 240 Thr Gly Gln Phe Leu Thr Phe Arg Glu Ile Asn Gly Met Thr Gly Leu 245 250 255 Asn Gly Ser Ile Gln Gln Ile Thr Val Ile Ser Pro Phe Ser Phe Ser 260 265 270 Ile Gly Asp Thr Thr Glu Leu Glu Pro Tyr Leu His Gly Gly Ile Ala 275 280 285 Val Gln Val Lys Thr Pro Lys Thr Val Phe Phe Glu Ser Leu Glu Arg 290 295 300 Gln Leu Lys His Pro Lys Cys Leu Ile Val Asp Phe Ser Asn Pro Glu 305 310 315 320 Ala Pro Leu Glu Ile His Thr Ala Met Leu Ala Leu Asp Gln Phe Gln 325 330 335 Glu Lys Tyr Ser Arg Lys Pro Asn Val Gly Cys Gln Gln Asp Ser Glu 340 345 350 Glu Leu Leu Lys Leu Ala Thr Ser Ile Ser Glu Thr Leu Glu Glu Lys 355 360 365 Pro Asp Val Asn Ala Asp Ile Val His Trp Leu Ser Trp Thr Ala Gln 370 375 380 Gly Phe Leu Ser Pro Leu Ala Ala Ala Val Gly Gly Val Ala Ser Gln 385 390 395 400 Glu Val Leu Lys Ala Val Thr Gly Lys Phe Ser Pro Leu Cys Gln Trp 405 410 415 Leu Tyr Leu Glu Ala Ala Asp Ile Val Glu Ser Leu Gly Lys Pro Glu 420 425 430 Cys Glu Glu Phe Leu Pro Arg Gly Asp Arg Tyr Asp Ala Leu Arg Ala 435 440 445 Cys Ile Gly Asp Thr Leu Cys Gln Lys Leu Gln Asn Leu Asn Ile Phe 450 455 460 Leu Val Gly Cys Gly Ala Ile Gly Cys Glu Met Leu Lys Asn Phe Ala 465 470 475 480 Leu Leu Gly Val Gly Thr Ser Lys Glu Lys Gly Met Ile Thr Val Thr 485 490 495 Asp Pro Asp Leu Ile Glu Lys Ser Asn Leu Asn Arg Gln Phe Leu Phe 500 505 510 Arg Pro His His Ile Gln Lys Pro Lys Ser Tyr Thr Ala Ala Asp Ala 515 520 525 Thr Leu Lys Ile Asn Ser Gln Ile Lys Ile Asp Ala His Leu Asn Lys 530 535 540 Val Cys Pro Thr Thr Glu Thr Ile Tyr Asn Asp Glu Phe Tyr Thr Lys 545 550 555 560 Gln Asp Val Ile Ile Thr Ala Leu Asp Asn Val Glu Ala Arg Arg Tyr 565 570 575 Val Asp Ser Arg Cys Leu Ala Asn Leu Arg Pro Leu Leu Asp Ser Gly 580 585 590 Thr Met Gly Thr Lys Gly His Thr Glu Val Ile Val Pro His Leu Thr 595 600 605 Glu Ser Tyr Asn Ser His Arg Asp Pro Pro Glu Glu Glu Ile Pro Phe 610 615 620 Cys Thr Leu Lys Ser Phe Pro Ala Ala Ile Glu His Thr Ile Gln Trp 625 630 635 640 Ala Arg Asp Lys Phe Glu Ser Ser Phe Ser His Lys Pro Ser Leu Phe 645 650 655 Asn Lys Phe Trp Gln Thr Tyr Ser Ser Ala Glu Glu Val Leu Gln Lys 660 665 670 Ile Gln Ser Gly His Ser Leu Glu Gly Cys Phe Gln Val Ile Lys Leu 675 680 685 Leu Ser Arg Arg Pro Arg Asn Trp Ser Gln Cys Val Glu Leu Ala Arg 690 695 700 Leu Lys Phe Glu Lys Tyr Phe Asn His Lys Ala Leu Gln Leu Leu His 705 710 715 720 Cys Phe Pro Leu Asp Ile Arg Leu Lys Asp Gly Ser Leu Phe Trp Gln 725 730 735 Ser Pro Lys Arg Pro Pro Ser Pro Ile Lys Phe Asp Leu Asn Glu Pro 740 745 750 Leu His Leu Ser Phe Leu Gln Asn Ala Ala Lys Leu Tyr Ala Thr Val 755 760 765 Tyr Cys Ile Pro Phe Ala Glu Glu Asp Leu Ser Ala Asp Ala Leu Leu 770 775 780 Asn Ile Leu Ser Glu Val Lys Ile Gln Glu Phe Lys Pro Ser Asn Lys 785 790 795 800 Val Val Gln Thr Asp Glu Thr Ala Arg Lys Pro Asp His Val Pro Ile 805 810 815 Ser Ser Glu Asp Glu Arg Asn Ala Ile Phe Gln Leu Glu Lys Ala Ile 820 825 830 Leu Ser Asn Glu Ala Thr Lys Ser Asp Leu Gln Met Ala Val Leu Ser 835 840 845 Phe Glu Lys Asp Asp Asp His Asn Gly His Ile Asp Phe Ile Thr Ala 850 855 860 Ala Ser Asn Leu Arg Ala Lys Met Tyr Ser Ile Glu Pro Ala Asp Arg 865 870 875 880 Phe Lys Thr Lys Arg Ile Ala Gly Lys Ile Ile Pro Ala Ile Ala Thr 885 890 895 Thr Thr Ala Thr Val Ser Gly Leu Val Ala Leu Glu Met Ile Lys Val 900 905 910 Thr Gly Gly Tyr Pro Phe Glu Ala Tyr Lys Asn Cys Phe Leu Asn Leu 915 920 925 Ala Ile Pro Ile Val Val Phe Thr Glu Thr Thr Glu Val Arg Lys Thr 930 935 940 Lys Ile Arg Asn Gly Ile Ser Phe Thr Ile Trp Asp Arg Trp Thr Val 945 950 955 960 His Gly Lys Glu Asp Phe Thr Leu Leu Asp Phe Ile Asn Ala Val Lys 965 970 975 Glu Lys Tyr Gly Ile Glu Pro Thr Met Val Val Gln Gly Val Lys Met 980 985 990 Leu Tyr Val Pro Met Met Pro Gly His Ala Lys Arg Leu Lys Leu Thr 995 1000 1005 Met His Lys Leu Val Lys Pro Thr Thr Glu Lys Lys Tyr Val Asp Leu 1010 1015 1020 Thr Val Ser Phe Ala Pro Asp Ile Asp Gly Asp Glu Asp Leu Pro Gly 1025 1030 1035 1040 Pro Pro Val Arg Tyr Tyr Phe Ser His Asp Thr Asp 1045 1050 <210> 3 <211> 1065 <212> DNA <213> USE1 cDNA sequence <400> 3 atggcggaga gtccgactga ggaggcggca acggcgggcg ccggggcggc gggccccggg 60 gcgagcagcg ttgctggtgt tgttggcgtt agcggcagcg gcggcgggtt cgggccgcct 120 ttcctgccgg atgtgtgggc ggcggcggcg gcagcgggcg gggccggggg cccggggagc 180 ggcctggctc cgctgcccgg gctcccgccc tcagccgctg cccacggggc cgcgctgctt 240 agccactggg accccacgct cagctccgac tgggacggcg agcgcaccgc gccgcagtgt 300 ctactccgga tcaagcggga tatcatgtcc atttataagg agcctcctcc aggaatgttc 360 gttgtacctg atactgttga catgactaag attcatgcat tgatcacagg cccatttgac 420 actccttatg aagggggttt cttcctgttc gtgtttcggt gtccgcccga ctatcccatc 480 cacccacctc gggtcaaact gatgacaacg ggcaataaca cagtgaggtt taaccccaac 540 ttctaccgca atgggaaagt ctgcttgagt attctaggta catggactgg acctgcctgg 600 agcccagccc agagcatctc ctcagtgctc atctctatcc agtccctgat gactgagaac 660 ccctatcaca atgagcccgg ctttgaacag gagagacatc caggagacag caaaaactat 720 aatgaatgta tccggcacga gaccatcaga gttgcagtct gtgacatgat ggaaggaaag 780 tgtccctgtc ctgaacccct acgaggggtg atggagaagt cctttctgga gtattacgac 840 ttctacgagg tggcctgcaa agatcgcctg caccttcaag gccaaactat gcaggaccct 900 tttggagaga agcggggcca ctttgactac cagtccctct tgatgcgcct gggactgata 960 cgtcagaaag tgctggagag gctccataat gagaatgcag aaatggactc tgatagcagt 1020 tcatctggga cagagacaga ccttcatggg agcctgaggg tttag 1065 <210> 4 <211> 354 <212> PRT <213> USE1 Protein sequence <400> 4 Met Ala Glu Ser Pro Thr Glu Glu Ala Ala Thr Ala Gly Ala Gly Ala 1 5 10 15 Ala Gly Pro Gly Ala Ser Ser Ala Gly Val Val Gly Val Ser Gly 20 25 30 Ser Gly Gly Gly Phe Gly Pro Pro Phe Leu Pro Asp Val Trp Ala Ala 35 40 45 Ala Ala Ala Ala Gly Aly Gly Aly Aly Gly Aly Aly Aly Aly 50 55 60 Leu Pro Gly Leu Pro Pro Ser Ala Ala Ala Ala Leu Leu 65 70 75 80 Ser His Trp Asp Pro Thr Leu Ser Ser Asp Trp Asp Gly Glu Arg Thr 85 90 95 Ala Pro Gln Cys Leu Leu Arg Ile Lys Arg Asp Ile Met Ser Ile Tyr 100 105 110 Lys Glu Pro Pro Gly Met Phe Val Val Pro Asp Thr Val Asp Met 115 120 125 Thr Lys Ile His Ala Leu Ile Thr Gly Pro Phe Asp Thr Pro Tyr Glu 130 135 140 Gly Gly Phe Phe Leu Phe Val Phe Arg Cys Pro Pro Asp Tyr Pro Ile 145 150 155 160 His Pro Pro Arg Val Lys Leu Met Thr Thr Gly Asn Asn Thr Val Arg 165 170 175 Phe Asn Pro Asn Phe Tyr Arg Asn Gly Lys Val Cys Leu Ser Ile Leu 180 185 190 Gly Thr Trp Thr Gly Pro Ala Trp Ser Pro Ala Gln Ser Ser Ser Ser 195 200 205 Val Leu Ile Ser Ile Gln Ser Leu Met Thr Glu Asn Pro Tyr His Asn 210 215 220 Glu Pro Gly Phe Glu Gln Glu Arg His Pro Gly Asp Ser Lys Asn Tyr 225 230 235 240 Asn Glu Cys Ile Arg His Glu Thr Ile Arg Val Ala Val Cys Asp Met 245 250 255 Met Glu Gly Lys Cys Pro Cys Pro Glu Pro Leu Arg Gly Val Met Glu 260 265 270 Lys Ser Phe Leu Glu Tyr Tyr Asp Phe Tyr Glu Val Ala Cys Lys Asp 275 280 285 Arg Leu His Leu Gln Gly Gln Thr Met Gln Asp Pro Phe Gly Glu Lys 290 295 300 Arg Gly His Phe Asp Tyr Gln Ser Leu Leu Met Arg Leu Gly Leu Ile 305 310 315 320 Arg Gln Lys Val Leu Glu Arg Leu His Asn Glu Asn Ala Glu Met Asp 325 330 335 Ser Asp Ser Ser Ser Gly Thr Glu Thr Asp Leu His Gly Ser Leu 340 345 350 Arg Val

Claims

A composition for diagnosing lung cancer, comprising an agent for measuring the expression level of mRNA of Uba6 (Ubiquitin-like modifier activating enzyme 6) or Use1 (Uba6 specific E2) gene or a protein thereof.

The method according to claim 1,
Wherein the agent for measuring the expression level of the gene mRNA comprises a primer that specifically binds to the Uba6 or Use1 gene.

The method according to claim 1,
Wherein the agent for measuring the expression level of the gene mRNA comprises a probe specifically binding to the Uba6 or Use1 gene.

The method according to claim 1,
Wherein the agent that measures the level of expression of said protein comprises an antibody specific for Uba6 or Use1 protein.

A kit for the diagnosis of lung cancer comprising the composition according to any one of claims 1 to 4.

6. The method of claim 5,
Wherein the kit is an RT-PCR kit, a DNA chip kit, or a protein chip kit.

(a) measuring the expression level or the amount of protein of the Uba6 or Use1 gene present in the biological sample; And
(b) comparing the measurement result of step (a) with the expression amount or protein amount of the Uba6 or Use1 gene of the normal control sample, thereby providing information for prediction and diagnosis of lung cancer.

8. The method of claim 7,
Wherein the biological sample is selected from the group consisting of tissue, cells, blood, serum, plasma, saliva, and urine.

8. The method of claim 7,
The measurement can be carried out using reverse transcriptase-polymerase chain reaction, real time-polymerase chain reaction, Western blot, northern blot, enzyme-linked immunosorbent assay (ELISA) A radioimmunoassay (RIA), a radioimmunoassay, and an immunoprecipitation assay.

8. The method of claim 7,
And determining that lung cancer has occurred if the expression amount or protein amount of Uba6 or Use1 gene present in said biological sample is increased as compared with that of a control.