KR101245112B1 - Radiation sensitive marker comprising APRIN gene or expression protein thereof, or protein regulated by expression-decreased APRIN gene - Google Patents

Abstract

The present invention relates to a radiation-sensitive marker comprising the APRIN gene or its expression protein, or a protein regulated by the reduction of gene expression, and to a medical use thereof, wherein the radiation sensitivity of the cell is increased when the expression amount of the APRIN gene is decreased. As a result, more cells are killed, and the expression of Chk2 and p-ATM decreases as the expression of APRIN gene decreases as a cause of the increase of cell death, thereby reducing the expression of the APRIN gene or its expression protein and APRIN gene. By using Chk2 and p-ATM, which are proteins whose expression is reduced, as markers for evaluating radiation sensitivity, a composition for enhancing radiation sensitivity can be developed using these inhibitors. Furthermore, by combining radiation therapy with reduced expression of APRIN, Chk2 or p-ATM, cancer cells can be suppressed by increasing the sensitivity of cancer cells to radiation, thereby improving radiotherapy effects in various cancer diseases such as lung cancer or osteosarcoma. Can be promoted.

Description

A radiation sensitive marker comprising APRIN gene or expression protein, or protein regulated by expression-decreased APRIN gene}

The present invention relates to radiation sensitive markers comprising the APRIN gene or its expression protein and medical use thereof.

Cancer treatments can be broadly divided into surgery, radiation therapy and chemotherapy. The treatment shows a cure rate of about 50%. Currently, about 35% of cancer patients in Korea and about 50% in the United States are receiving radiation therapy, and the number of cancer patients receiving radiation therapy in Korea is increasing every year. As a result, the importance of radiation therapy in the treatment of cancer is also increasing.

Radiation therapy is currently known as an essential treatment method for various types of cancers, but radiation resistance of cancer cells and damage to normal tissues during high-dose radiation treatments have become a problem of radiation therapy due to poor treatment efficiency.

Therefore, studies on enhancing agents for increasing the efficiency of such radiation treatments continue to progress, apoptosis stimulating agents that stimulate a variety of secondary signaling materials in the body or substances that stimulate cell death by stimulating cancer cell specific receptors, cells Research is conducted on periodic disturbances and transcription factor inhibitors.

Taxol and 5-FU (5-fluorouracil), which are currently known as anticancer agents, are known to enhance radiation treatment efficiency through co-administration.

However, when the anticancer agents used to enhance the radiotherapy effect in combination with radiation treatment, the toxicity of the anticancer agent, namely inflammation, gastrointestinal disorders, nausea, vomiting, diarrhea, etc. There is a limitation to use because it can appear complex.

In order to maximize the effect of radiation therapy to effectively treat various cancer diseases, the above problems have been solved, and development of radiation sensitive agents with minimal side effects is required.

On the other hand, APRIN is androgen-induced proliferation inhibitor (A ndrogen-induced Pr oliferation In hibitor) from were the name origin AS3; is also known as (androgen shutoff 3 AS3) or PDS5B (PDS5 regulator of cohesion maintenance homolog B), prostate It has been found to be a protein necessary for androgen-induced growth inhibition in cells. It is also known to combine with cohesion complexes to maintain sister chromatid cohesion when cell division and meiosis occur.

Accordingly, the present inventors found that when the expression level of the APRIN gene is decreased, the radiation sensitivity of the cell increases, thereby causing more cells to die, and the expression of Chk2 and p-ATM as the expression level of the APRIN gene decreases due to the increase in cell death. APRIN gene or its expression protein, Chk2 and p-ATM, proteins whose expression is reduced by decreasing the expression of the APRIN gene, are used as markers for evaluating radiation sensitivity, and at the same time, their inhibitors are used to determine the amount of radiation. The present invention has been completed by discovering that a composition for enhancing sensitivity can be developed.

It is an object of the present invention to provide a radiation-sensitive marker comprising the APRIN gene or its expression protein, Chk2 and p-ATM, which are proteins whose expression is regulated according to the expression reduction of the APRIN gene.

Another object of the present invention is to provide a radiation-sensitive diagnostic kit comprising APRIN gene or its expression protein, Chk2 and p-ATM, which are proteins whose expression is regulated according to a decrease in expression of the APRIN gene.

In addition, another object of the present invention is to enhance radiation sensitivity including any one inhibitor selected from the group consisting of APRIN gene or its expression protein, Chk2 and p-ATM, the protein whose expression is regulated according to the expression decrease of the APRIN gene. It is to provide a composition for.

In addition, another object of the present invention is an APRIN gene or its expression protein, a substance for enhancing radiation sensitivity using a radiation sensitive marker selected from the group consisting of Chk2 and p-ATM, the protein whose expression is controlled according to the expression decrease of the APRIN gene. It is to provide a screening method of.

In order to achieve the above object, the present invention is any one or more selected from the group consisting of APRIN gene or its expression protein encoding the amino acid sequence of SEQ ID NO: 1, and a protein whose expression is regulated according to the expression decrease of the APRIN gene It provides a radiation-sensitive marker comprising a.

The protein regulated by decreasing the expression of the APRIN gene is a protein whose expression is reduced by decreasing the expression of the APRIN gene, any one selected from the group consisting of Chk2 (SEQ ID NO: 2) and p-ATM (ATM: SEQ ID NO: 3) Or two or more.

That is, the present invention may be any one or two or more selected from the group consisting of APRIN gene encoding the amino acid sequence of SEQ ID NO: 1 or its expression protein, Chk2 and p-ATM which are proteins whose expression is reduced by decreasing the expression of the APRIN gene. It provides a radiation-sensitive marker comprising.

In addition, the present invention is one or more selected from the group consisting of APRIN gene encoding the amino acid sequence of SEQ ID NO: 1 or its expression protein, Chk2 and p-ATM which is a protein whose expression is reduced by a decrease in the expression of the APRIN gene. It provides a radiation sensitive diagnostic kit comprising.

According to the present invention, it was confirmed that radiation sensitivity is increased by reducing the expression of APRIN by treating APRIN inhibitor such as APRIN shRNA to human lung cancer cell line or human osteosarcoma cell line.

The treatment of human lung cancer cell lines or human osteosarcoma cell lines with APRIN inhibitors such as APRIN shRNA reduced the expression of APRIN and increased radiation sensitivity, confirming that more cancer cells die. As a cause of this apoptosis, cell cycle checkpoint defects and abnormal radiation-derived DNA damage signal transduction were identified, and in particular, the expression of Chk2 and p-ATM decreased with decreasing APRIN expression, which led to the APRIN gene. It can be used as a radiation-sensitive marker due to decreased expression.

The diagnostic kit of the present invention may include a sense and antisense primer of the APRIN gene, a probe complementary to the APRIN gene, or may include an antibody specific for the APRIN, Chk2 or p-ATM protein.

The present invention also provides a radiation-sensitive diagnostic microarray comprising the APRIN gene encoding the amino acid sequence of SEQ ID NO: 1.

In the present invention, "diagnosis" means identifying the presence or characteristic of a pathological condition. For the purposes of the present invention, the diagnosis is to determine the sensitivity to radiation therapy in cancer cells, such as lung cancer cells or osteosarcoma cells. In one embodiment, the present invention measures mRNA expression level of APRIN gene or expression level of APRIN, Chk2 or p-ATM protein from a biological sample obtained from an individual and compares it with the expression level of normal control sample for cancer disease. Radiation sensitivity can be diagnosed during radiation therapy.

In the present invention, "mRNA expression level measurement" is to measure the amount of mRNA in the process of confirming the presence and expression of mRNA of the APRIN gene in a biological sample to diagnose radiation sensitivity. Analytical methods for this purpose include, but are not limited to, RT-PCR, competitive RT-PCR, Real-time RT-PCR, Northern blotting, and DNA chips. It is not.

Through the above detection methods, it is possible to compare the mRNA expression level in the normal control group and the mRNA expression level in the biological sample obtained from the individual, and to determine whether the expression level of the APRIN gene to the mRNA is increased in the radiotherapy treatment for cancer disease. Diagnosis of radiation sensitivity can be made. mRNA expression level measurement may preferably be carried out using the RT-PCR method using a primer specific for the APRIN gene. RT-PCR is a method introduced to analyze RNA by P. Seeburg (Cold Spring Harb Symp Quant Biol 1986, Pt 1: 669-677). The cDNA obtained by reverse transcription of mRNA is amplified and analyzed by PCR. At this time, the primer pair prepared specifically for the APRIN gene is used in the amplification step, and electrophoresis after RT-PCR confirms the band pattern and the thickness of the APRIN gene, thereby confirming the mRNA expression and expression level of the APRIN gene and the normal control group. By comparing with, it is possible to easily diagnose the radiation sensitivity.

A "primer" of the present invention is a nucleic acid sequence having a short free 3 'hydroxyl group, which forms a complementary template and base pair and serves as a starting point for template strand copying. it means. Primers can initiate DNA synthesis in the presence of four different nucleotide triphosphates and reagents for polymerization (ie, DNA polymerase or reverse transcriptase) at appropriate buffers and temperatures. The primers of the invention can be sense and antisense nucleic acids having 7 to 50 nucleotide sequences as APRIN gene specific primers, and as long as they do not change the basic properties of the primers that serve as the starting point for DNA synthesis, Can be mixed.

Nucleic acid sequences of the invention can also be modified using a label that can provide a detectable signal directly or indirectly. Examples of labels include radioisotopes, fluorescent molecules, biotin, and the like.

In the present invention, "measurement of protein expression level" refers to a process for confirming the presence and expression level of APRIN, Chk2 or p-ATM protein in a biological sample in order to diagnose radiation sensitivity, and for APRIN, Chk2 or p-ATM protein. The amount of protein can be determined by using an antibody that binds specifically.

The protein expression level may be preferably measured by using an enzyme-linked immunosorbent assay (ELISA). ELISA is a direct ELISA using a labeled antibody that recognizes an antigen attached to a solid support, an indirect ELISA using a labeled antibody that recognizes a capture antibody in a complex of antibodies that recognize an antigen attached to a solid support, attached to a solid support Direct sandwich ELISA using another labeled antibody that recognizes the antigen in the antibody-antigen complex, a labeled antibody that recognizes the antibody after reacting with another antibody that recognizes the antigen in the complex of the antigen with the antibody attached to the solid support Various ELISA methods include indirect sandwich ELISA using secondary antibodies. More preferably, the antibody is enzymatically developed by attaching the antibody to the solid support, reacting the sample, and then attaching a labeled antibody that recognizes the antigen of the antigen-antibody complex, or to an antibody that recognizes the antigen of the antigen-antibody complex. It can be detected by a sandwich ELISA method in which the labeled secondary antibody is attached and developed enzymatically, and the degree of complex formation of the APRIN, Chk2 or p-ATM protein and the antibody can be confirmed to confirm radiation sensitivity.

Also preferably, Western blot analysis using one or more antibodies against APRIN, Chk2 or p-ATM protein can be used. The entire protein is isolated from the sample, electrophoresed to separate the protein according to size, and then transferred to a nitrocellulose membrane to react with the antibody. The amount of the generated antigen-antibody complex can be confirmed by radiation sensitivity by confirming the amount of APRIN, Chk2 or p-ATM protein by using a labeled antibody.

The detection method may be performed by examining the expression level of APRIN, Chk2 or p-ATM in the normal control group and the expression level thereof in the biological sample, and the mRNA or protein level may be absolute (eg, μg / Ml) or relative (e.g., the relative intensity of the signal).

"Antibody" in the present invention means a specific protein molecule directed against an antigenic site. For the purposes of the present invention, an antibody means an antibody that specifically binds to an APRIN, Chk2 or p-ATM protein, and includes both polyclonal antibodies, monoclonal antibodies and recombinant antibodies, and APRIN, Chk2 or p-ATM proteins. Antibodies to can be readily prepared using techniques well known in the art. Antibodies of the present invention can be prepared as monoclonal antibodies or polyclonal antibodies according to conventional methods known in the art, using these antibodies known enzyme linked immunosorbent assay (ELISA), radiation Diagnostics can be made by confirming that APRIN, Chk2 or p-ATM protein is expressed in a biological sample by methods such as radioimmunoassay (RIA), sandwich assay, western blotting or immunoblotting on polyacryl gel. Can be.

The microarray according to the present invention can be easily prepared by a manufacturing method commonly used in the art using the APRIN gene.

In addition, the present invention includes any one inhibitor selected from the group consisting of APRIN gene encoding the amino acid sequence of SEQ ID NO: 1 or its expression protein, Chk2 and p-ATM is a protein whose expression is reduced by a decrease in the expression of the APRIN gene. It provides a composition for enhancing radiation sensitivity.

The inhibitor may enhance sensitivity of cancer cells to radiation, and the cancer cells may be any one selected from lung cancer cells or osteosarcoma cells.

The inhibitor may be any one selected from antisense nucleic acids or small interfering RNAs that complementarily bind to mRNA of the APRIN gene, and compounds that bind to APRIN, Chk2 or p-ATM proteins, It may be any one selected from the group consisting of peptides and antibodies.

In one embodiment, the antisense nucleic acid of the present invention can be introduced into a cell to enhance the radiation sensitivity of cancer cells, where "introduction" introduces foreign DNA into the cell by transfection or transduction. Means that. Transfections include sugars such as calcium phosphate-DNA co-precipitation, DEAE-dextran-mediated transfection, polybrene-mediated transfection, electroshock, microinjection, liposome fusion, lipofectamine and protoplast fusion. It can be carried out by various methods known in the art. Transduction refers to the transfer of genes into cells using viral or viral vector particles by means of infection.

In the present invention, "radiation sensitivity enhancement" means to improve the sensitivity of the cells to radiation in the treatment of diseases using radiation. Through this, the radiation treatment efficiency can be increased. In particular, when the cancer treatment is performed in parallel, the radiation sensitivity of the cancer cells is enhanced, and the killing effect and the proliferation inhibitory effect of the cancer cells can be obtained.

The composition for enhancing radiation sensitivity of the present invention may further include a pharmaceutically acceptable carrier, and may be formulated with the carrier.

By "pharmaceutically acceptable carrier" is meant a carrier or diluent that does not stimulate the organism and does not inhibit the biological activity and properties of the administered compound. Examples of the pharmaceutical carrier which is acceptable for the composition to be formulated into a liquid solution include sterilized and sterile water suitable for the living body such as saline, sterilized water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, One or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like.

The composition for enhancing radiation sensitivity of the present invention can be applied in any dosage form, can be prepared in oral or parenteral dosage form, and can be formulated in unit dosage form for ease of administration and uniformity of dosage. Pharmaceutical formulations of the invention may be oral, rectal, nasal, topical (including the cheek and sublingual), subcutaneous, vaginal or parenteral (intramuscular, subcutaneous). And forms suitable for administration by inhalation or insufflation.

The oral dosage form may be formulated, for example, in tablets, troches, lozenges, water-soluble or oily suspensions, prepared powders or granules, emulsions, hard or soft capsules, syrups or elixirs. For formulation into tablets and capsules, lactose, saccharose, sorbitol, mannitol, starch, amylopectin, binders such as cellulose or gelatin, excipients such as dicalcium phosphate, disintegrants such as corn starch or sweet potato starch, stearic acid masne It may include a lubricating oil such as calcium, calcium stearate, sodium stearyl fumarate or polyethylene glycol wax, and in the case of a capsule, it may further contain a liquid carrier such as fatty oil in addition to the above-mentioned materials.

In addition, the parenteral formulation may be formulated for injection such as subcutaneous injection, intravenous injection or intramuscular injection, suppository injection method, or aerosol for inhalation through the respiratory system. To formulate injectable formulations, the compositions of the present invention may be mixed in water with stabilizers or buffers to prepare solutions or suspensions, which may be formulated for unit administration of ampoules or vials. For infusion into suppositories, it may be formulated as a rectal composition such as suppositories or body enema including conventional suppository bases such as cocoa butter or other glycerides. When formulated for spraying such as aerosols, a propellant or the like may be combined with the additives to disperse the dispersed dispersion or wet powder.

In addition, the present invention includes any one inhibitor selected from the group consisting of APRIN gene encoding the amino acid sequence of SEQ ID NO: 1 or its expression protein, Chk2 and p-ATM is a protein whose expression is reduced by a decrease in the expression of the APRIN gene. It provides a method for enhancing the radiation sensitivity of cancer cells using a composition for enhancing radiation sensitivity. In addition, there is provided a radiation therapy for the treatment of cancer comprising the step of administering the composition of the present invention.

By “administration” in the present invention is meant introducing the composition of the invention to a patient in any suitable way, including for example by delivery of an antisense nucleic acid molecule by viral or nonviral technology. When the composition of the present invention is introduced into cancer cells, the radiation sensitivity is enhanced by lowering the expression level or inhibiting the activity of APRIN.

The route of administration of the composition of the present invention can be administered via various routes orally or parenterally, as long as it can reach the target tissue, and specifically, oral, rectal, topical, intravenous, intraperitoneal, intramuscular, intraarterial It may be administered in a conventional manner via transdermal, nasal, inhalation, intraocular or intradermal routes. Preferably it is administered topically to the tissue.

Radiation therapy for the treatment of cancer of the present invention comprises administering a therapeutically effective amount of the composition for enhancing radiation sensitivity of the present invention. The therapeutically effective amount means an amount that effectively enhances the sensitivity of the tumor in cancer cells to radiation. It will be apparent to those skilled in the art that the appropriate total daily dose may be determined by the practitioner within the scope of sound medical judgment. The specific therapeutically effective amount for a particular patient may be based on the specific composition, including the type and severity of the reaction to be achieved, whether or not other agents are used in some cases, the age, body weight, general health, sex and diet, time of administration, It is desirable to apply differently depending on various factors including the route of administration and the rate of release of the composition, the duration of treatment, and the radiation dose to be irradiated and similar factors well known in the pharmaceutical art. Therefore, the effective amount of the composition for enhancing radiation sensitivity suitable for the purpose of the present invention is preferably determined in consideration of the above-mentioned matters. In addition, in some cases, by using a known anticancer agent in combination with the radiation sensitivity enhancing composition of the present invention can increase the anticancer effect including the radiation treatment effect.

In addition, the present invention provides a method of treating a biological sample comprising the steps of: treating any compound with a biological sample; Identifying the expression level of any one selected from the group consisting of APRIN gene or its expression protein encoding the amino acid sequence of SEQ ID NO: 1 from the biological sample, Chk2 and p-ATM, a protein whose expression is reduced by a decrease in the expression of the APRIN gene Making; And comparing the expression level with an expression level of an APRIN gene or an expression protein thereof in a normal control group not treated with any compound, thereby providing a compound screening method for enhancing sensitivity to radiation in cancer cells.

According to the present invention, when the expression level of the APRIN gene decreases, the radiation sensitivity of the cells increases, thereby killing more cells, and the expression of Chk2 and p-ATM due to the decrease in the expression amount of the APRIN gene as a cause of the increase of cell death. APRIN gene or its expression protein, Chk2 and p-ATM, proteins whose expression is reduced by decreasing the expression of the APRIN gene, are used as markers for evaluating radiation sensitivity, and at the same time, their inhibitors are used to determine the amount of radiation. A composition for enhancing sensitivity can be developed. In particular, the Chk2 and p-ATM are useful for evaluating radiation sensitivity in cancer cell lines with reduced APRIN expression. Furthermore, by combining radiation therapy with reduced expression of APRIN, Chk2 or p-ATM, cancer cells can be suppressed by increasing the sensitivity of cancer cells to radiation, thereby improving radiotherapy effects in various cancer diseases such as lung cancer or osteosarcoma. Can be promoted.

Figure 1 shows the increase in radiation sensitivity according to the decrease in the expression of APRIN,
Figure 2 examines the effect of cell proliferation after irradiation with reduced expression of APRIN,
Figure 3 shows the change in G2 / M checkpoint with reduced expression of APRIN,
4 shows the S-phase change according to the decrease in the expression of APRIN,
Figure 5 shows the radiation-induced DNA damage signaling changes with decreasing expression of APRIN.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the present invention is not limited by these examples.

Example 1 Cell Line Treatment

The human lung cancer cell line NCI-H460 cells and the human osteosarcoma cell line U2OS cells were dispensed in 2 × 10 ⁴ cells in a 24-well plate and incubated at 37 ° C. and 5% CO ₂ for 24 hours, followed by shRNA plasmid and control for APRIN. Lentiviruses containing shRNA plasmids were purchased from Santa Cruz (sc-61984-v) and infected with each. After 24 hours of infection, the culture medium was replaced for 48 hours, and then infected cells were re-inoculated into 4-8 wells in 24-well plates for 24 hours. After 24 hours, the cells were replaced with a culture solution containing 2 μg / ml of puromycin, and then the cultures were changed every 2-3 days, and only surviving cells were selected and used in subsequent experiments.

Example 2 Examination of Radiation Sensitivity Change with Reduction of APRIN Expression

In order to confirm the radiation sensitivity of the cells according to the decrease in the expression of APRIN, a colony assay was performed. That is, H460 cells expressing the control shRNA and shAPRIN on a 60 mm plate were divided into 100, 200, 500, 1000, 2000 cells, and cultured for 24 hours. After 24 hours, each cell was irradiated with 0, 1, 3, 4 Gy and incubated for 10-14 days. When colonies of the appropriate size were formed, they were stained with 0.4% trypan blue and the number of colonies was measured.

As a result, as shown in FIG. 1, the cells in which APRIN expression was reduced by using the shAPRIN plasmid were confirmed to have reduced colony formation as compared to the control cells. Thus, cells with reduced APRIN expression increased radiation sensitivity compared to control cells. I could see that.

Example 3 Examination of Cell Proliferation after Irradiation with Reduction of APRIN Expression

When the expression of APRIN was reduced, WST-1 analysis was performed to confirm the cell proliferation change after irradiation. That is, 1 × 10 ³ U2OS cells and H460 cells expressing the control shRNA and shAPRIN in 96-well plate were dispensed and cultured for 24 hours. After 24 hours of incubation, 5 Gy of radiation was applied, and 1/10 volume of WST-1 reagent was added and reacted at 37 ° C. for 1 hour, and then the absorbance was measured at 450 nm and set to 0 days. After 1, 2, 4, and 6 days Absorbance at 450 nm was measured by adding 1/10 volume of WST-1 reagent.

As a result, as shown in FIG. 2, when the U2OS and H460 cells were irradiated with radiation, the proliferation of cells was reduced as compared with the unirradiated cells. In the case of H460 cells, the irradiated or non-irradiated cells showed more proliferation of cells with reduced APRIN expression, and the difference was not significantly affected by irradiation. However, in the case of U2OS cells, the cells that inhibited the expression of APRIN showed almost the same cell proliferation as the control group when irradiated with radiation. Therefore, the effect of APRIN expression inhibition on the cell proliferation by radiation was different depending on the cell lines. It was found out.

Example 4 Examining Changes in G2 / M Checkpoints with Reduction of APRIN Expression

Ser10 phosphorylation of intracellular Histone H3, an M phase marker, was irradiated with 0.5 Gy of control cells and APRIN-reduced cells to confirm the change in cell cycle checkpoints when irradiated with reduced APRIN expression. The change of was confirmed. That is, H460 cells expressing the control shRNA and shAPRIN on a 60 mm plate were divided into 6 × 10 ⁵ cells and incubated for 24 hours. After incubation with 0.5 Gy of radiation, the cells were recovered at 0, 0.5, 1 and 2 hours and fixed with 90% cold methanol. The immobilized cells were washed with 1 × PBS and reacted for 1 hour by diluting 1/100 of the FITC-conjugated p-Histone H3 (Ser10) antibody, followed by 25 μg / ml of RNase A and 25 μg / ml of propidium iodine. Id was added and FACS analysis was performed.

As a result, as shown in FIG. 3, when 30 minutes after irradiation, the decrease in the expression of phospho-H3 (Ser10) of the cells that reduced the expression of APRIN was less than that of the control group. It was found that less G2 / M arrest occurred.

<Example 5> Examination of S-phase change by decreasing expression of APRIN

When irradiated cells with reduced expression of APRIN were treated with BrdU to control and cells with reduced expression of APRIN to confirm the change in S-phase, the amount of BrdU was checked after irradiation after irradiation. That is, H460 cells expressing the control shRNA and shAPRIN on a 100 mm plate were divided into 1 × 10 ⁶ cells and cultured for 24 hours. One hour prior to irradiation, 10 μM of bromodeoxyuridine (BrdU) was pretreated and irradiated with 5 Gy of radiation, cells were recovered after 0, 3, 8, and 24 hours and fixed in 70% ethanol. Fixed cells were washed with 1 × PBS and then treated with 1 ml of 2 M HCl (+ 0.5% BSA) for 20 minutes. After the cells were recovered by centrifugation, 1 ml of 0.1 M Na-borate (pH 8.5) was treated for 2 minutes and centrifuged to recover the cells. After washing with PBS once, the FITC-conjugated BrdU antibody was diluted 1/100 and reacted for 1 hour. Then, 25 μg / ml of RNase A and 25 μg / ml of propiodium iodide were added and FACS analysis was performed.

As a result, as shown in FIG. 4, the difference between the two cells could not be confirmed after 3 hours of irradiation, but at 8 and 24 hours, the S-phase was reduced in the cells in which APRIN expression was reduced compared to the control group. We can see that more exists. As a result, the decrease in the expression of APRIN increased the radiation sensitivity by affecting the regulation of S-phase.

<Example 6> Examination of radiation-induced DNA damage signaling changes with decreased expression of APRIN

To investigate the effect of decreased expression of APRIN on signal transduction, DNA damage-related signals were detected at 1 and 4 hours after irradiation of 2 Gy and 5 Gy of control cells and cells with reduced expression of APRIN. Confirmed. That is, H460 cells expressing the control shRNA and shAPRIN were divided into 100 mm plates with 1 × 10 ⁶ cells and cultured for 24 hours. The cultured cells were irradiated with 2 Gy and 5 Gy, and then washed twice with cold 1 × PBS after 1 and 4 hours, and scraped off the plate to collect the cells. The cells were lysed with RIPA buffer (50 mM Tris-Cl (pH8.0), 150 mM NaCl, 1% NP-40, 0.1% SDS, 0.1% deoxycholate, protease inhibitor, phosphatase inhibitor) Intracellular protein was extracted. The extracted protein was quantified by Bradford method, and the expression level of the protein was confirmed by Western blot analysis.

As a result, as shown in FIG. 5, the phosphorylation of Chk2 and ATM was reduced in the cells which decreased the expression of APRIN compared to the control group, and it was confirmed that the phosphorylation of p53 was slightly decreased compared to the control group, although weaker than these. Irradiation with 2 Gy also reduced the expression of γ-H2AX, a marker of DNA damage. From these results, it was determined that the reduction of APRIN affects the cellular radiosensitivity by affecting the DNA damage signaling such as Chk2, p53, ATM.

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereby. something to do. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

<110> KOREA INSTITUTE OF RADIOLOGICAL & MEDICAL SCIENCES <120> Radiation sensitive marker comprising APRIN gene or expression          protein according, or protein regulated by expression-decreased          APRIN gene <130> DP-2010-0592 <160> 3 <170> Kopatentin 1.71 <210> 1 <211> 302 <212> PRT <213> Homo sapiens <400> 1 Ile Lys Gln Thr Lys Asp Ala Gln Gly Pro Asp Asp Ala Lys Met Asn   1 5 10 15 Glu Lys Leu Tyr Thr Val Cys Asp Val Ala Met Asn Ile Met Ser              20 25 30 Lys Ser Thr Thr Tyr Ser Leu Glu Ser Pro Lys Asp Pro Val Leu Pro          35 40 45 Ala Arg Phe Phe Thr Gln Pro Asp Lys Asn Phe Ser Asn Thr Lys Asn      50 55 60 Tyr Leu Pro Pro Glu Met Lys Ser Phe Phe Thr Pro Gly Lys Pro Lys  65 70 75 80 Thr Thr Asn Val Leu Gly Ala Val Asn Lys Pro Leu Ser Ser Ala Gly                  85 90 95 Lys Gln Ser Gln Thr Lys Ser Ser Arg Met Glu Thr Val Ser Asn Ala             100 105 110 Ser Ser Ser Ser Asn Pro Ser Ser Pro Gly Arg Ile Lys Gly Arg Leu         115 120 125 Asp Ser Ser Glu Met Asp His Ser Glu Asn Glu Asp Tyr Thr Met Ser     130 135 140 Ser Pro Leu Pro Gly Lys Lys Ser Asp Lys Arg Asp Asp Ser Asp Leu 145 150 155 160 Ser Glu Leu Glu Lys Pro Arg Gly Arg Lys Lys Thr Pro Val Thr Glu                 165 170 175 Gln Glu Glu Lys Leu Gly Met Asp Asp Leu Thr Lys Leu Val Gln Glu             180 185 190 Gln Lys Pro Lys Gly Ser Gln Arg Ser Arg Lys Arg Gly His Thr Ala         195 200 205 Ser Glu Ser Asp Glu Gln Gln Trp Pro Glu Glu Lys Arg Leu Lys Glu     210 215 220 Asp Ile Leu Glu Asn Glu Asp Glu Gln Asn Ser Pro Pro Lys Lys Gly 225 230 235 240 Lys Arg Gly Arg Pro Pro Lys Pro Leu Gly Gly Gly Thr Pro Lys Glu                 245 250 255 Glu Pro Thr Met Lys Thr Ser Lys Lys Gly Ser Lys Lys Lys Ser Gly             260 265 270 Pro Pro Ala Pro Glu Glu Glu Glu Glu Glu Glu Arg Gln Ser Gly Asn         275 280 285 Thr Glu Gln Lys Ser Lys Ser Lys Gln His Arg Val Ser Arg     290 295 300 <210> 2 <211> 543 <212> PRT <213> Homo sapiens <400> 2 Met Ser Arg Glu Ser Asp Val Glu Ala Gln Gln Ser His Gly Ser Ser   1 5 10 15 Ala Cys Ser Gln Pro His Gly Ser Val Thr Gln Ser Gln Gly Ser Ser              20 25 30 Ser Gln Ser Gln Gly Ile Ser Ser Ser Ser Thr Ser Thr Met Pro Asn          35 40 45 Ser Ser Gln Ser Ser His Ser Ser Ser Gly Thr Leu Ser Ser Leu Glu      50 55 60 Thr Val Ser Thr Gln Glu Leu Tyr Ser Ile Pro Glu Asp Gln Glu Pro  65 70 75 80 Glu Asp Gln Glu Pro Glu Glu Pro Thr Pro Ala Pro Trp Ala Arg Leu                  85 90 95 Trp Ala Leu Gln Asp Gly Phe Ala Asn Leu Glu Cys Val Asn Asp Asn             100 105 110 Tyr Trp Phe Gly Arg Asp Lys Ser Cys Glu Tyr Cys Phe Asp Glu Pro         115 120 125 Leu Leu Lys Arg Thr Asp Lys Tyr Arg Thr Tyr Ser Lys Lys His Phe     130 135 140 Arg Ile Phe Arg Glu Val Gly Pro Lys Asn Ser Tyr Ile Ala Tyr Ile 145 150 155 160 Glu Asp His Ser Gly Asn Gly Thr Phe Val Asn Thr Glu Leu Val Gly                 165 170 175 Lys Gly Lys Arg Arg Pro Leu Asn Asn Asn Ser Glu Ile Ala Leu Ser             180 185 190 Leu Ser Arg Asn Lys Val Phe Val Phe Phe Asp Leu Thr Val Asp Asp         195 200 205 Gln Ser Val Tyr Pro Lys Ala Leu Arg Asp Glu Tyr Ile Met Ser Lys     210 215 220 Thr Leu Gly Ser Gly Ala Cys Gly Glu Val Lys Leu Ala Phe Glu Arg 225 230 235 240 Lys Thr Cys Lys Lys Val Ala Ile Lys Ile Ile Ser Lys Arg Lys Phe                 245 250 255 Ala Ile Gly Ser Ala Arg Glu Ala Asp Pro Ala Leu Asn Val Glu Thr             260 265 270 Glu Ile Glu Ile Leu Lys Lys Leu Asn His Pro Cys Ile Ile Lys Ile         275 280 285 Lys Asn Phe Phe Asp Ala Glu Asp Tyr Tyr Ile Val Leu Glu Leu Met     290 295 300 Glu Gly Gly Glu Leu Phe Asp Lys Val Val Gly Asn Lys Arg Leu Lys 305 310 315 320 Glu Ala Thr Cys Lys Leu Tyr Phe Tyr Gln Met Leu Leu Ala Val Gln                 325 330 335 Tyr Leu His Glu Asn Gly Ile Ile His Arg Asp Leu Lys Pro Glu Asn             340 345 350 Val Leu Leu Ser Ser Gln Glu Glu Asp Cys Leu Ile Lys Ile Thr Asp         355 360 365 Phe Gly His Ser Lys Ile Leu Gly Glu Thr Ser Leu Met Arg Thr Leu     370 375 380 Cys Gly Thr Pro Thr Tyr Leu Ala Pro Glu Val Leu Val Ser Val Gly 385 390 395 400 Thr Ala Gly Tyr Asn Arg Ala Val Asp Cys Trp Ser Leu Gly Val Ile                 405 410 415 Leu Phe Ile Cys Leu Ser Gly Tyr Pro Pro Phe Ser Glu His Arg Thr             420 425 430 Gln Val Ser Leu Lys Asp Gln Ile Thr Ser Gly Lys Tyr Asn Phe Ile         435 440 445 Pro Glu Val Trp Ala Glu Val Ser Glu Lys Ala Leu Asp Leu Val Lys     450 455 460 Lys Leu Leu Val Val Asp Pro Lys Ala Arg Phe Thr Thr Glu Glu Ala 465 470 475 480 Leu Arg His Pro Trp Leu Gln Asp Glu Asp Met Lys Arg Lys Phe Gln                 485 490 495 Asp Leu Leu Ser Glu Glu Asn Glu Ser Thr Ala Leu Pro Gln Val Leu             500 505 510 Ala Gln Pro Ser Thr Ser Arg Lys Arg Pro Arg Glu Gly Glu Ala Glu         515 520 525 Gly Ala Glu Thr Thr Lys Arg Pro Ala Val Cys Ala Ala Val Leu     530 535 540 <210> 3 <211> 3055 <212> PRT <213> Homo sapiens <400> 3 Met Ser Leu Val Leu Asn Asp Leu Leu Ile Cys Cys Arg Gln Leu Glu   1 5 10 15 His Asp Arg Ala Thr Glu Arg Lys Lys Glu Val Glu Lys Phe Lys Arg              20 25 30 Leu Ile Arg Asp Pro Glu Thr Ile Lys His Leu Asp Arg His Ser Asp          35 40 45 Ser Lys Gln Gly Lys Tyr Leu Asn Trp Asp Ala Val Arg Phe Leu Gln      50 55 60 Lys Tyr Ile Gln Lys Glu Thr Glu Cys Leu Arg Ile Ala Lys Pro Asn  65 70 75 80 Val Ser Ala Ser Thr Gln Ala Ser Arg Gln Lys Lys Met Gln Glu Ile                  85 90 95 Ser Ser Leu Val Lys Tyr Phe Ile Lys Cys Ala Asn Arg Arg Ala Pro             100 105 110 Arg Leu Lys Cys Gln Glu Leu Leu Asn Tyr Ile Met Asp Thr Val Lys         115 120 125 Asp Ser Ser Asn Gly Ala Ile Tyr Gly Ala Asp Cys Ser Asn Ile Leu     130 135 140 Leu Lys Asp Ile Leu Ser Val Arg Lys Tyr Trp Cys Glu Ile Ser Gln 145 150 155 160 Gln Gln Trp Leu Glu Leu Phe Ser Val Tyr Phe Arg Leu Tyr Leu Lys                 165 170 175 Pro Ser Gln Asp Val His Arg Val Leu Val Ala Arg Ile Ile His Ala             180 185 190 Val Thr Lys Gly Cys Cys Ser Gln Thr Asp Gly Leu Asn Ser Lys Phe         195 200 205 Leu Asp Phe Phe Ser Lys Ala Ile Gln Cys Ala Arg Gln Glu Lys Ser     210 215 220 Ser Ser Gly Leu Asn His Ile Leu Ala Ala Leu Thr Ile Phe Leu Lys 225 230 235 240 Thr Leu Ala Val Asn Phe Arg Ile Arg Val Cys Glu Leu Gly Asp Glu                 245 250 255 Ile Leu Pro Thr Leu Leu Tyr Ile Trp Thr Gln His Arg Leu Asn Asp             260 265 270 Ser Leu Lys Glu Val Ile Ile Glu Leu Phe Gln Leu Gln Ile Tyr Ile         275 280 285 His His Pro Lys Gly Ala Lys Thr Gln Glu Lys Gly Ala Tyr Glu Ser     290 295 300 Thr Lys Trp Arg Ser Ile Leu Tyr Asn Leu Tyr Asp Leu Leu Val Asn 305 310 315 320 Glu Ile Ser His Ile Gly Ser Arg Gly Lys Tyr Ser Ser Gly Phe Arg                 325 330 335 Asn Ile Ala Val Lys Glu Asn Leu Ile Glu Leu Met Ala Asp Ile Cys             340 345 350 His Gln Val Phe Asn Glu Asp Thr Arg Ser Leu Glu Ile Ser Gln Ser         355 360 365 Tyr Thr Thr Thr Gln Arg Glu Ser Ser Asp Tyr Ser Val Pro Cys Lys     370 375 380 Arg Lys Lys Ile Glu Leu Gly Trp Glu Val Ile Lys Asp His Leu Gln 385 390 395 400 Lys Ser Gln Asn Asp Phe Asp Leu Val Pro Trp Leu Gln Ile Ala Thr                 405 410 415 Gln Leu Ile Ser Lys Tyr Pro Ala Ser Leu Pro Asn Cys Glu Leu Ser             420 425 430 Pro Leu Leu Met Ile Leu Ser Gln Leu Leu Pro Gln Gln Arg His Gly         435 440 445 Glu Arg Thr Pro Tyr Val Leu Arg Cys Leu Thr Glu Val Ala Leu Cys     450 455 460 Gln Asp Lys Arg Ser Asn Leu Glu Ser Ser Gln Lys Ser Asp Leu Leu 465 470 475 480 Lys Leu Trp Asn Lys Ile Trp Cys Ile Thr Phe Arg Gly Ile Ser Ser                 485 490 495 Glu Gln Ile Gln Ala Glu Asn Phe Gly Leu Leu Gly Ala Ile Ile Gln             500 505 510 Gly Ser Leu Val Glu Val Asp Arg Glu Phe Trp Lys Leu Phe Thr Gly         515 520 525 Ser Ala Cys Arg Pro Ser Cys Pro Ala Val Cys Cys Leu Thr Leu Ala     530 535 540 Leu Thr Thr Ser Ile Val Pro Gly Thr Val Lys Met Gly Ile Glu Gln 545 550 555 560 Asn Met Cys Glu Val Asn Arg Ser Phe Ser Leu Lys Glu Ser Ile Met                 565 570 575 Lys Trp Leu Leu Phe Tyr Gln Leu Glu Gly Asp Leu Glu Asn Ser Thr             580 585 590 Glu Val Pro Pro Ile Leu His Ser Asn Phe Pro His Leu Val Leu Glu         595 600 605 Lys Ile Leu Val Ser Leu Thr Met Lys Asn Cys Lys Ala Ala Met Asn     610 615 620 Phe Phe Gln Ser Val Pro Glu Cys Glu His His Gln Lys Asp Lys Glu 625 630 635 640 Glu Leu Ser Phe Ser Glu Val Glu Glu Leu Phe Leu Gln Thr Thr Phe                 645 650 655 Asp Lys Met Asp Phe Leu Thr Ile Val Arg Glu Cys Gly Ile Glu Lys             660 665 670 His Gln Ser Ser Ile Gly Phe Ser Val His Gln Asn Leu Lys Glu Ser         675 680 685 Leu Asp Arg Cys Leu Leu Gly Leu Ser Glu Gln Leu Leu Asn Asn Tyr     690 695 700 Ser Ser Glu Ile Thr Asn Ser Glu Thr Leu Val Arg Cys Ser Arg Leu 705 710 715 720 Leu Val Gly Val Leu Gly Cys Tyr Cys Tyr Met Gly Val Ile Ala Glu                 725 730 735 Glu Glu Ala Tyr Lys Ser Glu Leu Phe Gln Lys Ala Lys Ser Leu Met             740 745 750 Gln Cys Ala Gly Glu Ser Ile Thr Leu Phe Lys Asn Lys Thr Asn Glu         755 760 765 Glu Phe Arg Ile Gly Ser Leu Arg Asn Met Met Gln Leu Cys Thr Arg     770 775 780 Cys Leu Ser Asn Cys Thr Lys Lys Ser Pro Asn Lys Ile Ala Ser Gly 785 790 795 800 Phe Phe Leu Arg Leu Leu Thr Ser Lys Leu Met Asn Asp Ile Ala Asp                 805 810 815 Ile Cys Lys Ser Leu Ala Ser Phe Ile Lys Lys Pro Phe Asp Arg Gly             820 825 830 Glu Val Glu Ser Met Glu Asp Asp Thr Asn Gly Asn Leu Met Glu Val         835 840 845 Glu Asp Gln Ser Ser Met Asn Leu Phe Asn Asp Tyr Pro Asp Ser Ser     850 855 860 Val Ser Asp Ala Asn Glu Pro Gly Glu Ser Gln Ser Thr Ile Gly Ala 865 870 875 880 Ile Asn Pro Leu Ala Glu Glu Tyr Leu Ser Lys Gln Asp Leu Leu Phe                 885 890 895 Leu Asp Met Leu Lys Phe Leu Cys Leu Cys Val Thr Thr Ala Gln Thr             900 905 910 Asn Thr Val Ser Phe Arg Ala Ala Asp Ile Arg Arg Lys Leu Leu Met         915 920 925 Leu Ile Asp Ser Ser Thr Leu Glu Pro Thr Lys Ser Leu His Leu His     930 935 940 Met Tyr Leu Met Leu Leu Lys Glu Leu Pro Gly Glu Glu Tyr Pro Leu 945 950 955 960 Pro Met Glu Asp Val Leu Glu Leu Leu Lys Pro Leu Ser Asn Val Cys                 965 970 975 Ser Leu Tyr Arg Arg Asp Gln Asp Val Cys Lys Thr Ile Leu Asn His             980 985 990 Val Leu His Val Val Lys Asn Leu Gly Gln Ser Asn Met Asp Ser Glu         995 1000 1005 Asn Thr Arg Asp Ala Gln Gly Gln Phe Leu Thr Val Ile Gly Ala Phe    1010 1015 1020 Trp His Leu Thr Lys Glu Arg Lys Tyr Ile Phe Ser Val Arg Met Ala 1025 1030 1035 1040 Leu Val Asn Cys Leu Lys Thr Leu Leu Glu Ala Asp Pro Tyr Ser Lys                1045 1050 1055 Trp Ala Ile Leu Asn Val Met Gly Lys Asp Phe Pro Val Asn Glu Val            1060 1065 1070 Phe Thr Gln Phe Leu Ala Asp Asn His His Gln Val Arg Met Leu Ala        1075 1080 1085 Ala Glu Ser Ile Asn Arg Leu Phe Gln Asp Thr Lys Gly Asp Ser Ser    1090 1095 1100 Arg Leu Leu Lys Ala Leu Pro Leu Lys Leu Gln Gln Thr Ala Phe Glu 1105 1110 1115 1120 Asn Ala Tyr Leu Lys Ala Gln Glu Gly Met Arg Glu Met Ser His Ser                1125 1130 1135 Ala Glu Asn Pro Glu Thr Leu Asp Glu Ile Tyr Asn Arg Lys Ser Val            1140 1145 1150 Leu Leu Thr Leu Ile Ala Val Val Leu Ser Cys Ser Pro Ile Cys Glu        1155 1160 1165 Lys Gln Ala Leu Phe Ala Leu Cys Lys Ser Val Lys Glu Asn Gly Leu    1170 1175 1180 Glu Pro His Leu Val Lys Lys Val Leu Glu Lys Val Ser Glu Thr Phe 1185 1190 1195 1200 Gly Tyr Arg Arg Leu Glu Asp Phe Met Ala Ser His Leu Asp Tyr Leu                1205 1210 1215 Val Leu Glu Trp Leu Asn Leu Gln Asp Thr Glu Tyr Asn Leu Ser Ser            1220 1225 1230 Phe Pro Phe Ile Leu Leu Asn Tyr Thr Asn Ile Glu Asp Phe Tyr Arg        1235 1240 1245 Ser Cys Tyr Lys Val Leu Ile Pro His Leu Val Ile Arg Ser His Phe    1250 1255 1260 Asp Glu Val Lys Ser Ile Ala Asn Gln Ile Gln Glu Asp Trp Lys Ser 1265 1270 1275 1280 Leu Leu Thr Asp Cys Phe Pro Lys Ile Leu Val Asn Ile Leu Pro Tyr                1285 1290 1295 Phe Ala Tyr Glu Gly Thr Arg Asp Ser Gly Met Ala Gln Gln Arg Glu            1300 1305 1310 Thr Ala Thr Lys Val Tyr Asp Met Leu Lys Ser Glu Asn Leu Leu Gly        1315 1320 1325 Lys Gln Ile Asp His Leu Phe Ile Ser Asn Leu Pro Glu Ile Val Val    1330 1335 1340 Glu Leu Leu Met Thr Leu His Glu Pro Ala Asn Ser Ser Ala Ser Gln 1345 1350 1355 1360 Ser Thr Asp Leu Cys Asp Phe Ser Gly Asp Leu Asp Pro Ala Pro Asn                1365 1370 1375 Pro Pro His Phe Pro Ser His Val Ile Lys Ala Thr Phe Ala Tyr Ile            1380 1385 1390 Ser Asn Cys His Lys Thr Lys Leu Lys Ser Ile Leu Glu Ile Leu Ser        1395 1400 1405 Lys Ser Pro Asp Ser Tyr Gln Lys Ile Leu Leu Ala Ile Cys Glu Gln    1410 1415 1420 Ala Ala Glu Thr Asn Asn Val Tyr Lys Lys His Arg Ile Leu Lys Ile 1425 1430 1435 1440 Tyr His Leu Phe Val Ser Leu Leu Leu Lys Asp Ile Lys Ser Gly Leu                1445 1450 1455 Gly Gly Ala Trp Ala Phe Val Leu Arg Asp Val Ile Tyr Thr Leu Ile            1460 1465 1470 His Tyr Ile Asn Gln Arg Pro Ser Cys Ile Met Asp Val Ser Leu Arg        1475 1480 1485 Ser Phe Ser Leu Cys Cys Asp Leu Leu Ser Gln Val Cys Gln Thr Ala    1490 1495 1500 Val Thr Tyr Cys Lys Asp Ala Leu Glu Asn His Leu His Val Ile Val 1505 1510 1515 1520 Gly Thr Leu Ile Pro Leu Val Tyr Glu Gln Val Glu Val Gln Lys Gln                1525 1530 1535 Val Leu Asp Leu Leu Lys Tyr Leu Val Ile Asp Asn Lys Asp Asn Glu            1540 1545 1550 Asn Leu Tyr Ile Thr Ile Lys Leu Leu Asp Pro Phe Pro Asp His Val        1555 1560 1565 Val Phe Lys Asp Leu Arg Ile Thr Gln Gln Lys Ile Lys Tyr Ser Arg    1570 1575 1580 Gly Pro Phe Ser Leu Leu Glu Glu Ile Asn His Phe Leu Ser Val Ser 1585 1590 1595 1600 Val Tyr Asp Ala Leu Pro Leu Thr Arg Leu Glu Gly Leu Lys Asp Leu                1605 1610 1615 Arg Arg Gln Leu Glu Leu His Lys Asp Gln Met Val Asp Ile Met Arg            1620 1625 1630 Ala Ser Gln Asp Asn Pro Gln Asp Gly Ile Met Val Lys Leu Val Val        1635 1640 1645 Asn Leu Leu Gln Leu Ser Lys Met Ala Ile Asn His Thr Gly Glu Lys    1650 1655 1660 Glu Val Leu Glu Ala Val Gly Ser Cys Leu Gly Glu Val Gly Pro Ile 1665 1670 1675 1680 Asp Phe Ser Thr Ile Ala Ile Gln His Ser Lys Asp Ala Ser Tyr Thr                1685 1690 1695 Lys Ala Leu Lys Leu Phe Glu Asp Lys Glu Leu Gln Trp Thr Phe Ile            1700 1705 1710 Met Leu Thr Tyr Leu Asn Asn Thr Leu Val Glu Asp Cys Val Lys Val        1715 1720 1725 Arg Ser Ala Ala Val Thr Cys Leu Lys Asn Ile Leu Ala Thr Lys Thr    1730 1735 1740 Gly His Ser Phe Trp Glu Ile Tyr Lys Met Thr Thr Asp Pro Met Leu 1745 1750 1755 1760 Ala Tyr Leu Gln Pro Phe Arg Thr Ser Arg Lys Lys Phe Leu Glu Val                1765 1770 1775 Pro Arg Phe Asp Lys Glu Asn Pro Phe Glu Gly Leu Asp Asp Ile Asn            1780 1785 1790 Leu Trp Ile Pro Leu Ser Glu Asn His Asp Ile Trp Ile Lys Thr Leu        1795 1800 1805 Thr Cys Ala Phe Leu Asp Ser Gly Gly Thr Lys Cys Glu Ile Leu Gln    1810 1815 1820 Leu Leu Lys Pro Met Cys Glu Val Lys Thr Asp Phe Cys Gln Thr Val 1825 1830 1835 1840 Leu Pro Tyr Leu Ile His Asp Ile Leu Leu Gln Asp Thr Asn Glu Ser                1845 1850 1855 Trp Arg Asn Leu Leu Ser Thr His Val Gln Gly Phe Phe Thr Ser Cys            1860 1865 1870 Leu Arg His Phe Ser Gln Thr Ser Arg Ser Thr Thr Pro Ala Asn Leu        1875 1880 1885 Asp Ser Glu Ser Glu His Phe Phe Arg Cys Cys Leu Asp Lys Lys Ser    1890 1895 1900 Gln Arg Thr Met Leu Ala Val Val Asp Tyr Met Arg Arg Gln Lys Arg 1905 1910 1915 1920 Pro Ser Ser Gly Thr Ile Phe Asn Asp Ala Phe Trp Leu Asp Leu Asn                1925 1930 1935 Tyr Leu Glu Val Ala Lys Val Ala Gln Ser Cys Ala Ala His Phe Thr            1940 1945 1950 Ala Leu Leu Tyr Ala Glu Ile Tyr Ala Asp Lys Lys Ser Met Asp Asp        1955 1960 1965 Gln Glu Lys Arg Ser Leu Ala Phe Glu Glu Gly Ser Gln Ser Thr Thr    1970 1975 1980 Ile Ser Ser Leu Ser Glu Lys Ser Lys Glu Glu Thr Gly Ile Ser Leu 1985 1990 1995 2000 Gln Asp Leu Leu Leu Glu Ile Tyr Arg Ser Ile Gly Glu Pro Asp Ser                2005 2010 2015 Leu Tyr Gly Cys Gly Gly Gly Lys Met Leu Gln Pro Ile Thr Arg Leu            2020 2025 2030 Arg Thr Tyr Glu His Glu Ala Met Trp Gly Lys Ala Leu Val Thr Tyr        2035 2040 2045 Asp Leu Glu Thr Ala Ile Pro Ser Ser Thr Arg Gln Ala Gly Ile Ile    2050 2055 2060 Gln Ala Leu Gln Asn Leu Gly Leu Cys His Ile Leu Ser Val Tyr Leu 2065 2070 2075 2080 Lys Gly Leu Asp Tyr Glu Asn Lys Asp Trp Cys Pro Glu Leu Glu Glu                2085 2090 2095 Leu His Tyr Gln Ala Ala Trp Arg Asn Met Gln Trp Asp His Cys Thr            2100 2105 2110 Ser Val Ser Lys Glu Val Glu Gly Thr Ser Tyr His Glu Ser Leu Tyr        2115 2120 2125 Asn Ala Leu Gln Ser Leu Arg Asp Arg Glu Phe Ser Thr Phe Tyr Glu    2130 2135 2140 Ser Leu Lys Tyr Ala Arg Val Lys Glu Val Glu Glu Met Cys Lys Arg 2145 2150 2155 2160 Ser Leu Glu Ser Val Tyr Ser Leu Tyr Pro Thr Leu Ser Arg Leu Gln                2165 2170 2175 Ala Ile Gly Glu Leu Glu Ser Ile Gly Glu Leu Phe Ser Arg Ser Val            2180 2185 2190 Thr His Arg Gln Leu Ser Glu Val Tyr Ile Lys Trp Gln Lys His Ser        2195 2200 2205 Gln Leu Leu Lys Asp Ser Asp Phe Ser Phe Gln Glu Pro Ile Met Ala    2210 2215 2220 Leu Arg Thr Val Ile Leu Glu Ile Leu Met Glu Lys Glu Met Asp Asn 2225 2230 2235 2240 Ser Gln Arg Glu Cys Ile Lys Asp Ile Leu Thr Lys His Leu Val Glu                2245 2250 2255 Leu Ser Ile Leu Ala Arg Thr Phe Lys Asn Thr Gln Leu Pro Glu Arg            2260 2265 2270 Ala Ile Phe Gln Ile Lys Gln Tyr Asn Ser Val Ser Cys Gly Val Ser        2275 2280 2285 Glu Trp Gln Leu Glu Glu Ala Gln Val Phe Trp Ala Lys Lys Glu Gln    2290 2295 2300 Ser Leu Ala Leu Ser Ile Leu Lys Gln Met Ile Lys Lys Leu Asp Ala 2305 2310 2315 2320 Ser Cys Ala Ala Asn Asn Pro Ser Leu Lys Leu Thr Tyr Thr Glu Cys                2325 2330 2335 Leu Arg Val Cys Gly Asn Trp Leu Ala Glu Thr Cys Leu Glu Asn Pro            2340 2345 2350 Ala Val Ile Met Gln Thr Tyr Leu Glu Lys Ala Val Glu Val Ala Gly        2355 2360 2365 Asn Tyr Asp Gly Glu Ser Ser Asp Glu Leu Arg Asn Gly Lys Met Lys    2370 2375 2380 Ala Phe Leu Ser Leu Ala Arg Phe Ser Asp Thr Gln Tyr Gln Arg Ile 2385 2390 2395 2400 Glu Asn Tyr Met Lys Ser Ser Glu Phe Glu Asn Lys Gln Ala Leu Leu                2405 2410 2415 Lys Arg Ala Lys Glu Glu Val Gly Leu Leu Arg Glu His Lys Ile Gln            2420 2425 2430 Thr Asn Arg Tyr Thr Val Lys Val Gln Arg Glu Leu Glu Leu Asp Glu        2435 2440 2445 Leu Ala Leu Arg Ala Leu Lys Glu Asp Arg Lys Arg Phe Leu Cys Lys    2450 2455 2460 Ala Val Glu Asn Tyr Ile Asn Cys Leu Leu Ser Gly Glu Glu His Asp 2465 2470 2475 2480 Met Trp Val Phe Arg Leu Cys Ser Leu Trp Leu Glu Asn Ser Gly Val                2485 2490 2495 Ser Glu Val Asn Gly Met Met Lys Arg Asp Gly Met Lys Ile Pro Thr            2500 2505 2510 Tyr Lys Phe Leu Pro Leu Met Tyr Gln Leu Ala Ala Arg Met Gly Thr        2515 2520 2525 Lys Met Met Gly Gly Leu Gly Phe His Glu Val Leu Asn Asn Leu Ile    2530 2535 2540 Ser Arg Ile Ser Met Asp His Pro His His Thr Leu Phe Ile Ile Leu 2545 2550 2555 2560 Ala Leu Ala Asn Ala Asn Arg Asp Glu Phe Leu Thr Lys Pro Glu Val                2565 2570 2575 Ala Arg Arg Ser Arg Ile Thr Lys Asn Val Pro Lys Gln Ser Ser Gln            2580 2585 2590 Leu Asp Glu Asp Arg Thr Glu Ala Ala Asn Arg Ile Ile Cys Thr Ile        2595 2600 2605 Arg Ser Arg Arg Pro Gln Met Val Arg Ser Val Glu Ala Leu Cys Asp    2610 2615 2620 Ala Tyr Ile Ile Leu Ala Asn Leu Asp Ala Thr Gln Trp Lys Thr Gln 2625 2630 2635 2640 Arg Lys Gly Ile Asn Ile Pro Ala Asp Gln Pro Ile Thr Lys Leu Lys                2645 2650 2655 Asn Leu Glu Asp Val Val Val Pro Thr Met Glu Ile Lys Val Asp His            2660 2665 2670 Thr Gly Glu Tyr Gly Asn Leu Val Thr Ile Gln Ser Phe Lys Ala Glu        2675 2680 2685 Phe Arg Leu Ala Gly Gly Val Asn Leu Pro Lys Ile Ile Asp Cys Val    2690 2695 2700 Gly Ser Asp Gly Lys Glu Arg Arg Gln Leu Val Lys Gly Arg Asp Asp 2705 2710 2715 2720 Leu Arg Gln Asp Ala Val Met Gln Gln Val Phe Gln Met Cys Asn Thr                2725 2730 2735 Leu Leu Gln Arg Asn Thr Glu Thr Arg Lys Arg Lys Leu Thr Ile Cys            2740 2745 2750 Thr Tyr Lys Val Val Pro Leu Ser Gln Arg Ser Gly Val Leu Glu Trp        2755 2760 2765 Cys Thr Gly Thr Val Pro Ile Gly Glu Phe Leu Val Asn Asn Glu Asp    2770 2775 2780 Gly Ala His Lys Arg Tyr Arg Pro Asn Asp Phe Ser Ala Phe Gln Cys 2785 2790 2795 2800 Gln Lys Lys Met Met Glu Val Gln Lys Lys Ser Phe Glu Glu Lys Tyr                2805 2810 2815 Glu Val Phe Met Asp Val Cys Gln Asn Phe Gln Pro Val Phe Arg Tyr            2820 2825 2830 Phe Cys Met Glu Lys Phe Leu Asp Pro Ala Ile Trp Phe Glu Lys Arg        2835 2840 2845 Leu Ala Tyr Thr Arg Ser Val Ala Thr Ser Ser Ile Val Gly Tyr Ile    2850 2855 2860 Leu Gly Leu Gly Asp Arg His Val Gln Asn Ile Leu Ile Asn Glu Gln 2865 2870 2875 2880 Ser Ala Glu Leu Val His Ile Asp Leu Gly Val Ala Phe Glu Gln Gly                2885 2890 2895 Lys Ile Leu Pro Thr Pro Glu Thr Val Pro Phe Arg Leu Thr Arg Asp            2900 2905 2910 Ile Val Asp Gly Met Gly Ile Thr Gly Val Glu Gly Val Phe Arg Arg        2915 2920 2925 Cys Cys Glu Lys Thr Met Glu Val Met Arg Asn Ser Gln Glu Thr Leu    2930 2935 2940 Leu Thr Ile Val Glu Val Leu Leu Tyr Asp Pro Leu Phe Asp Trp Thr 2945 2950 2955 2960 Met Asn Pro Leu Lys Ala Leu Tyr Leu Gln Gln Arg Pro Glu Asp Glu                2965 2970 2975 Thr Glu Leu His Pro Thr Leu Asn Ala Asp Asp Gln Glu Cys Lys Arg            2980 2985 2990 Asn Leu Ser Asp Ile Asp Gln Ser Phe Asn Lys Val Ala Glu Arg Val        2995 3000 3005 Leu Met Arg Leu Gln Glu Lys Leu Lys Gly Val Glu Glu Gly Thr Val    3010 3015 3020 Leu Ser Val Gly Gly Gln Val Asn Leu Leu Ile Gln Gln Ala Ile Asp 3025 3030 3035 3040 Pro Lys Asn Leu Ser Arg Leu Phe Pro Gly Trp Lys Ala Trp Val                3045 3050 3055

Claims (9)

A radiation sensitive marker composition comprising an APRIN gene or an expression protein thereof encoding an amino acid sequence of SEQ ID NO: 1 inversely proportional to radiation sensitivity. The method according to claim 1,
Further comprising a protein whose expression is reduced by decreasing the expression of the APRIN gene, wherein the protein is any one or two or more selected from the group consisting of Chk2 (SEQ ID NO: 2) and p-ATM (ATM: SEQ ID NO: 3) Phosphorus radiation sensitive marker composition. A radiation sensitive diagnostic kit comprising an APRIN gene encoding the amino acid sequence of SEQ ID NO: 1 or an expression protein thereof. A composition for enhancing radiation sensitivity comprising an inhibitor of the APRIN gene or its expression protein encoding the amino acid sequence of SEQ ID NO: 1. The composition of claim 4, wherein the inhibitor enhances the sensitivity of cancer cells to radiation. The composition for enhancing radiation sensitivity according to claim 5, wherein the cancer cells are any one selected from lung cancer cells or osteosarcoma cells. The composition of claim 4 or 5, wherein the inhibitor is any one selected from antisense nucleotides or small interfering RNAs complementarily binding to mRNA of the APRIN gene. The composition for enhancing radiation sensitivity according to claim 4 or 5, wherein the inhibitor is any one selected from the group consisting of a compound, a peptide, and an antibody complementarily binding to the APRIN protein. Treating any compound with a biological sample; Identifying the expression level of the APRIN gene or its expressed protein encoding the amino acid sequence of SEQ ID NO: 1 from the biological sample; And comparing the expression level with the expression level of the same gene or protein in the normal control group not treated with any compound.

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