KR102177320B1 - A biomarker for each dose of low dose radiation exposure, a diagnostic method using the biomarker, and a diagmositic kit including the biomarker - Google Patents
A biomarker for each dose of low dose radiation exposure, a diagnostic method using the biomarker, and a diagmositic kit including the biomarker Download PDFInfo
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Abstract
본 발명은 저선량방사선 피폭 선량별 바이오마커, 상기 바이오마커를 이용한 저선량방사선 피폭 진단방법 및 상기 바이오마커를 포함하는 저선량방사선 피폭 진단 키트에 관한 것으로서, microRNA-142-5p(miR-142-5p) 또는 microRNA-155-5p(miR-155-5p)를 포함한다.
상기와 같은 본 발명에 따르면, 마우스 저선량방사선 전신 노출에 의한 비장 및 골수에서 아직 보고된바 없는 새로운 miRNA에 대하여 발굴 및 제시함으로써, 의료 및 방사선작업종사 그리고 일반인의 100 mSv 이하의 저선량방사선의 방사선 피폭을 빠르고 효율적으로 진단할 수 있는 효과가 있다. The present invention relates to a biomarker for each dose of low-dose radiation, a method for diagnosing low-dose radiation using the biomarker, and a low-dose radiation exposure diagnostic kit comprising the biomarker, microRNA-142-5p (miR-142-5p) or microRNA-155-5p (miR-155-5p).
According to the present invention as described above, by discovering and presenting new miRNAs that have not yet been reported in the spleen and bone marrow caused by whole-body exposure to low-dose mice, medical and radiation workers and the general public are exposed to radiation of low-dose radiation of less than 100 mSv. There is an effect that can be quickly and efficiently diagnosed.
Description
본 발명은 저선량방사선 피폭 선량별 바이오마커, 상기 바이오마커를 이용한 진단방법 및 상기 바이오마커를 포함하는 진단 키트에 관한 것으로서, 더욱 상세하게는 저선량방사선 노출에 의해 특이적으로 발현이 증가하는 miRNA를 발굴하고 이를 이용하는 것에 관한 것이다.The present invention relates to a biomarker for each dose of low-dose radiation, a diagnostic method using the biomarker, and a diagnostic kit including the biomarker, and more particularly, to discover miRNAs whose expression is specifically increased by exposure to low-dose radiation. And using it.
지난 수십 년간 매우 낮은 수준의 이온화방사선이 인체의 건강에 미치는 영향을 정량화하려고 노력해왔다. 병원에서 방사선치료 또는 처치를 받거나 원자력산업 및 의료산업 근로자의 방사선노출로 발생하는 저선량방사선의 인체 영향에 대한 방사선 노출의 위험을 과학적으로 구체화하는 구체적 수치를 제시하는 것이 필요하다. Over the past decades, efforts have been made to quantify the effects of very low levels of ionizing radiation on human health. It is necessary to present specific values that scientifically specify the risk of radiation exposure to the human effects of low-dose radiation arising from radiation exposure of workers in the nuclear and medical industries, receiving radiation treatment or treatment in hospitals.
저선량방사선 노출에 대한 인체의 즉각적인 증상이나 영향은 관찰되고 있지는 않지만, 인체 영향에 대한 잠재적 불확실성과 위험성에 대하여 방사선 피폭량을 정확하게 측정 가능한 생물학적 특이 반응 지표의 개발이 필요하다.Although no immediate symptoms or effects of the human body have been observed on exposure to low-dose radiation, it is necessary to develop a biologically specific response indicator that can accurately measure the radiation exposure dose for potential uncertainty and risk of human effects.
microRNA(miRNA)는 수많은 유전자의 발현을 조절하는 짧은 단편(18-22개 뉴클레오티드 길이)의 non-coding RNA를 말하며, 유전자의 전사 후 단계에서 번역을 억제함으로써 타겟 mRNA를 코딩하는 유전자의 발현을 억제하는 조절자 역할을 한다. 최근에 다양한 miRNA들이 저선량방사선 노출에 의해 조절되며, 특히 miR-193b-3p 및 miR-134을 포함하는 몇몇 miRNA는 간, 뇌 등 다양한 기관에서 조절되는 것으로 보고된바 있다. microRNA (miRNA) refers to a short fragment (18-22 nucleotides in length) of non-coding RNA that controls the expression of many genes. It suppresses the expression of the gene encoding the target mRNA by inhibiting translation in the post-transcription stage of the gene. To act as a mediator. Recently, various miRNAs have been reported to be regulated by exposure to low-dose radiation, and in particular, some miRNAs, including miR-193b-3p and miR-134, have been reported to be regulated in various organs such as liver and brain.
따라서 본 발명은 이전에 공지된 저선량방사선 노출에 의해 조절되는 miRNA가 아닌, 마우스의 저선량방사선 전신 노출에 대한 비장 및 골수에서 아직 보고된바 없는 새로운 miRNA에 대하여 발굴 및 제시하고자 한다. Therefore, the present invention intends to discover and present new miRNAs that have not yet been reported in the spleen and bone marrow for systemic exposure to low-dose radiation in mice, not miRNAs controlled by previously known low-dose radiation exposure.
본 발명의 목적은, 마우스 저선량방사선 전신 노출에 의한 비장 및 골수에서 아직 보고된바 없는 새로운 miRNA에 대하여 발굴 및 제시함으로써, 의료 및 방사선작업종사 그리고 일반인의 100 mSv 이하의 저선량방사선의 방사선 피폭을 빠르고 효율적으로 진단할 수 있는 바이오진단 마커로 활용함에 있다.It is an object of the present invention to discover and present new miRNAs that have not yet been reported in the spleen and bone marrow by whole-body exposure to low-dose mice, so that radiation exposure of low-dose radiation of less than 100 mSv of medical and radiation workers and the general public can be quickly It is to be used as a biodiagnostic marker that can be efficiently diagnosed.
상기 목적을 달성하기 위하여, 본 발명은 microRNA-142-5p(miR-142-5p) 또는 microRNA-155-5p(miR-155-5p)를 포함하는 저선량방사선 피폭 선량별 바이오마커, 상기 바이오마커를 이용한 저선량방사선 피폭 진단방법 및 상기 바이오마커를 포함하는 저선량방사선 피폭 진단 키트를 제공한다.In order to achieve the above object, the present invention comprises a biomarker for each dose of low-dose radiation, including microRNA-142-5p (miR-142-5p) or microRNA-155-5p (miR-155-5p), the biomarker. A method for diagnosing low-dose radiation using and a kit for diagnosing low-dose radiation including the biomarker.
상기와 같은 본 발명에 따르면, 마우스 저선량방사선 전신 노출에 의한 비장 및 골수에서 아직 보고된바 없는 새로운 miRNA에 대하여 발굴 및 제시함으로써, 의료 및 방사선작업종사 그리고 일반인의 100 mSv 이하의 저선량방사선의 방사선 피폭을 빠르고 효율적으로 진단할 수 있는 효과가 있다.According to the present invention as described above, by discovering and presenting new miRNAs that have not yet been reported in the spleen and bone marrow caused by whole-body exposure to low-dose mice, medical and radiation workers and the general public are exposed to radiation of low-dose radiation of less than 100 mSv. There is an effect that can be quickly and efficiently diagnosed.
도 1은 C57BL/6 마우스의 비장에서 naive CD4+ T 세포를 분리하여 활성화시킨 CD4+ T 세포를 대조군으로 저선량방사선을 조사하여 24시간 후에 total RNA를 분리하여 miRNA microarray를 수행한 결과로, 대조군과 비교하여 저선량방사선에 의해 특이적으로 조절되는 (A) mir-142-5p, (B) mir-155-5p를 도시하였다.
도 2는 C57BL/6 마우스에 저선량방사선을 전신조사하고 4시간 후 골수세포를 분리하여 total RNA로부터 (A) mir-142-5p, (B) mir-155-5p의 발현량을 RT-PCR을 이용하여 확인하고, 이를 그래프로 도시하였다.
도 3은 인간 면역 T세포 (HuT78)에 저선량방사선을 조사하고 24시간 후 total RNA로부터 (A) mir-142-5p, (B) mir-155-5p의 발현량을 RT-PCR을 이용하여 확인하고, 이를 그래프로 도시하였다. 1 is a result of performing miRNA microarray by separating total RNA after 24 hours by irradiating low-dose radiation on CD4+ T cells activated by separating naive CD4+ T cells from the spleen of C57BL/6 mice as a control. (A) mir-142-5p and (B) mir-155-5p, which are specifically regulated by low-dose radiation, are shown.
FIG. 2 is a C57BL/6 mouse systemically irradiated with low-dose radiation, and after 4 hours, bone marrow cells were isolated, and the expression levels of (A) mir-142-5p, (B) mir-155-5p were measured from total RNA by RT-PCR. It was confirmed using, and this was shown in a graph.
3 is a human immune T cell (HuT78) irradiated with low-dose radiation and 24 hours later, the expression levels of (A) mir-142-5p and (B) mir-155-5p from total RNA were confirmed using RT-PCR. And, it is shown as a graph.
이하, 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명의 일 형태에 따라 microRNA-142-5p(miR-142-5p) 또는 microRNA-155-5p(miR-155-5p)를 포함하는 저선량방사선 피폭 선량별 바이오마커를 제공한다. According to one embodiment of the present invention, a biomarker for each dose of low-dose radiation is provided, including microRNA-142-5p (miR-142-5p) or microRNA-155-5p (miR-155-5p).
상기 microRNA-142-5p(miR-142-5p)는 저선량방사선 노출시 저선량방사선 비노출에 비해 특이적으로 발현이 증가할 수 있다. The microRNA-142-5p (miR-142-5p) may have a specific increase in expression when exposed to low-dose radiation compared to non-exposed to low-dose radiation.
상기 microRNA-155-5p(miR-155-5p)는 저선량방사선 노출시 저선량방사선 비노출에 비해 특이적으로 발현이 증가할 수 있다. The microRNA-155-5p (miR-155-5p) may specifically increase expression when exposed to low-dose radiation compared to non-exposed to low-dose radiation.
상기 저선량방사선은 10 내지 100 mSv의 선량이다. The low-dose radiation is a dose of 10 to 100 mSv.
본 발명의 다른 형태에 따라 저선량방사선 피폭 진단방법을 제공한다. According to another aspect of the present invention, a method for diagnosing low-dose radiation exposure is provided.
상기 저선량방사선 피폭 진단방법은 사람을 제외한 동물의 생체시료를 준비하는 단계, 상기 생체시료의 miRNA(microRNA)를 분석하는 단계 및 상기 miRNA(microRNA) 중에 miR-142-5p 또는 miR-155-5p를 저선량방사선에 비노출된 대조군과 비교하는 단계를 포함할 수 있다. The method for diagnosing low-dose radiation exposure includes preparing a biological sample of animals other than humans, analyzing miRNA (microRNA) of the biological sample, and using miR-142-5p or miR-155-5p in the miRNA (microRNA). It may comprise the step of comparing with a control group that is not exposed to low-dose radiation.
상기 생체시료는 혈액, 비장 또는 골수 일 수 있다. The biological sample may be blood, spleen or bone marrow.
상기 miRNA(microRNA)를 분석하는 방법은 정량 RT-PCR 분석 일 수 있다. The method of analyzing the miRNA (microRNA) may be quantitative RT-PCR analysis.
상기 대조군과 비교하는 단계는 miR-142-5p 또는 miR-155-5p가 대조군에 비해 특이적으로 발현이 증가되는 것을 확인하는 것 일 수 있다. The step of comparing with the control may be confirming that the expression of miR-142-5p or miR-155-5p is specifically increased compared to the control.
본 발명의 다른 형태에 따라 microRNA-142-5p(miR-142-5p) 또는 microRNA-155-5p(miR-155-5p)인 바이오마커를 포함하는 저선량방사선 피폭 진단 키트를 제공한다. According to another aspect of the present invention, a diagnostic kit for low-dose radiation exposure comprising a biomarker of microRNA-142-5p (miR-142-5p) or microRNA-155-5p (miR-155-5p) is provided.
상기 microRNA-142-5p(miR-142-5p)는 저선량방사선 노출시 저선량방사선 비노출에 비해 특이적으로 발현이 증가할 수 있다. The microRNA-142-5p (miR-142-5p) may have a specific increase in expression when exposed to low-dose radiation compared to non-exposed to low-dose radiation.
상기 microRNA-155-5p(miR-155-5p)는 저선량방사선 노출시 저선량방사선 비노출에 비해 특이적으로 발현이 증가할 수 있다. The microRNA-155-5p (miR-155-5p) may specifically increase expression when exposed to low-dose radiation compared to non-exposed to low-dose radiation.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail through examples. These examples are for illustrative purposes only, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not construed as being limited by these examples.
실시예 1. 마우스 비장에서 저선량방사선에 의해 특이적으로 반응하는 miRNA들의 확인 (miRNA microarray) Example 1. Identification of miRNAs reacting specifically by low-dose radiation in mouse spleen (miRNA microarray)
6주령 C57BL/6 마우스 수컷의 비장에서 naive CD4+ T 세포를 분리하기 위해 분리 키트 (isolation kit) 매뉴얼에 따라 음성선택법 (negative selection)에 의해 분리하였다. 분리된 CD4+ T 세포를 항-CD3 항체 (5ug/ml)와 항-CD28 항체 (5ug/ml)를 이용하여 1시간동안 활성화시킨 후, 저선량 방사선을 조사하였다. In order to isolate naive CD4+ T cells from the spleen of a 6-week-old male C57BL/6 mouse, the cells were isolated by negative selection according to the isolation kit manual. The isolated CD4+ T cells were activated for 1 hour with anti-CD3 antibody (5ug/ml) and anti-CD28 antibody (5ug/ml), and then irradiated with low-dose radiation.
저선량방사선 조사는 이온화방사선 (137Cs, 930 mGy/min, Grammacell 40 Exactor, Best Theratronics Ltd.)을 80% 차폐 처리하여 선량률 (186 mGy/min)로 4가지 선량별 (10, 25, 50, 100 mGy)로 실시하였다. 저선량방사선을 10, 25, 50, 100 mGy로 각각의 실험군에 조사하고 24시간 동안 배양 후 세포에서 total RNA를 분리하였다. 이때 저선량방사선을 조사하지 않은 활성화된 CD4+ T 세포를 대조군으로 사용하기 위해 total RNA를 분리하였다. Total RNA 분리는 Trizol 시약 (Invitrogen, Carlsbad, CA)을 사용하여 제조사의 방법에 따라 추출하였다. 추출된 각 RNA들은, 이바이오젠 (한국)에서 제공하는 miRNA 탐색기술인 miRNA micro-array 방법을 이용하여, 활성화된 CD4+ T 세포에서 저선량방사선이 조사되었을 때 변화되는 miRNA들을 검색하였다. For low-dose radiation irradiation, 80% of ionizing radiation (137Cs, 930 mGy/min, Grammacell 40 Exactor, Best Theratronics Ltd.) is shielded at a dose rate (186 mGy/min) for 4 doses (10, 25, 50, 100 mGy). ). Low-dose radiation was irradiated to each experimental group at 10, 25, 50, and 100 mGy, and after incubation for 24 hours, total RNA was isolated from the cells. At this time, total RNA was isolated to use activated CD4+ T cells that were not irradiated with low-dose radiation as a control. Total RNA was isolated using Trizol reagent (Invitrogen, Carlsbad, CA) and extracted according to the manufacturer's method. Each extracted RNA was searched for miRNAs that were changed when low-dose radiation was irradiated in activated CD4+ T cells using the miRNA micro-array method, a miRNA search technology provided by eBiogen (Korea).
분석 결과, 약 3100종류의 선량별로 발현 변화에 차이가 있는 miRNA를 확인 할 수 있었으며, 그중에 저선량방사선에 특이적으로 반응하는 miRNA를 도1에 도시하였다. As a result of the analysis, about 3100 types of miRNAs having a difference in expression change for each dose were identified, and among them, miRNAs that specifically respond to low-dose radiation are shown in FIG. 1.
도 1을 참고하면 2종의 miRNA(mir-142-5p. mir-155-5p)가 저선량방사선 10, 25, 50, 100 mGy의 노출에 의해 특이적으로 반응이 증가 되는 것이 확인 할 수 있었다. Referring to FIG. 1, it was confirmed that the reaction of two miRNAs (mir-142-5p. mir-155-5p) was specifically increased by exposure to low-dose radiation of 10, 25, 50, and 100 mGy.
실시예 2. 저선량방사선 전신조사된 마우스의 골수세포에서 저선량방사선 특이적 miRNA의 발현변화 검증 Example 2. Verification of changes in the expression of low-dose-specific miRNA in bone marrow cells of mice irradiated with low-dose radiation systemically
상기 실시예 1에서와 같이 방사선에 민감한 골수세포에서 확인된 2종의 miRNA를 추가적으로 확인 검증하기 위해 10주령 Balb/C 마우스 수컷에 저선량방사선(10, 25, 50 mGy)을 조사하였다. 저선량방사선 조사양에 따라 각 그룹당 3마리씩 전신 조사하여 4시간 후에 대퇴골(femurs) 및 경골(tibias)로부터 26 게이지 주사기 바늘을 이용하여 페니실린 (100 U/ml)과 스트렙토마이신 (100ug/ml)이 포한된 RPMI1640 배지를 뼈사이로 통과시키고 40 um nylon mesh (BD Biosciences, San Jose, CA)의 세포 여과기(strainer)을 사용하여 세포를 1차로 분리하였다. 추가적으로 적혈구를 제거하기 위해 RBC lysis 용액인 ACK lysing buffer (Lonza, Gampel, Switzerland)를 상온에서 5분 동안 처리하고, 차가운 1XPBS로 1회 세척하여 최종적으로 골수세포를 분리하였다. As in Example 1, low-dose radiation (10, 25, 50 mGy) was irradiated to males of 10-week-old Balb/C mice to additionally confirm and verify the two types of miRNAs identified in radiation-sensitive bone marrow cells. Depending on the amount of low-dose irradiation, 3 animals per group were whole-body irradiated and after 4 hours, penicillin (100 U/ml) and streptomycin (100 ug/ml) were contained from the femurs and tibias using a 26 gauge syringe needle. The RPMI1640 medium was passed through the bones and cells were first separated using a cell strainer of 40 um nylon mesh (BD Biosciences, San Jose, CA). In order to additionally remove red blood cells, ACK lysing buffer (Lonza, Gampel, Switzerland), an RBC lysis solution, was treated for 5 minutes at room temperature and washed once with cold 1XPBS to finally isolate bone marrow cells.
분리한 골수세포에서 Trizol 시약 (Invitrogen, Carlsbad, CA)를 사용하여 total RNA를 추출 하였고, cDNA는 miScript II RT Kit (Qiagen, Hilden, Germany)를 사용하여 합성하였다. 정량 PCR은 ABI PRISM 7500 Sequence Detection System (ABI, Foster city, CA) 및 miScript SYBR Green Kit (Qiagen, Hilden, Germany)를 사용하여 분석하였다. 이때 내부 대조군으로 RNU6-2을 사용하여 3번 반복하여 측정 하였으며, PCR 프라이머의 서열은 하기 표 1에 도시하였다. Total RNA was extracted from the isolated bone marrow cells using Trizol reagent (Invitrogen, Carlsbad, CA), and cDNA was synthesized using miScript II RT Kit (Qiagen, Hilden, Germany). Quantitative PCR was analyzed using an ABI PRISM 7500 Sequence Detection System (ABI, Foster city, CA) and miScript SYBR Green Kit (Qiagen, Hilden, Germany). At this time, the measurement was repeated 3 times using RNU6-2 as an internal control, and the sequence of the PCR primers is shown in Table 1 below.
분석 결과는 도 2에 도시하였다. The analysis results are shown in FIG. 2.
도 2를 참조하면, 도 1에서 확인된 바와 같이, 저선량방사선에 의해 miRNAs (mir-142-5p. mir-155-5p)가 유사하게 조절되는 것이 추가적으로 검증 되었다. Referring to FIG. 2, as confirmed in FIG. 1, it was additionally verified that miRNAs (mir-142-5p. mir-155-5p) are similarly regulated by low-dose radiation.
실시예3. 인간 면역세포 (HuT78)에서 저선량방사선 특이적 miRNA의 발현변화 검증 Example 3. Verification of changes in expression of low-dose-specific miRNA in human immune cells (HuT78)
상기 실시예 2에서 확인된 miRNAs (mir-142-5p. mir-155-5p)의 반응조절인자를 인간 세포주에서 확인하기 위해, 인간 피부 T 세포 림프종 (Cutaneous T Lymphocyte) 세포주 HuT78를 사용하여 저선량방사선에 의한 반응조절인자를 추가적으로 검증하고자 하였다. HuT78 세포주는 20% FBS, 100U/ml 페니실린 및 100ug/ml 스트렙토마이신 (Gibco, USA)이 첨가된 RPMI1640 배지에서 37℃, 5% CO2 조건에서 배양되었다. 저선량방사선 조사는 이온화방사선 (137Cs, 930 mGy/min, Grammacell 40 Exactor, Best Theratronics Ltd.)을 80% 차폐 처리하여 선량률 (186 mGy/min)로 4가지 선량별 (10, 25, 50, 100 mGy)로 시행하였다. 세포주 HuT78을 항-CD3 항체 (5ug/ml)와 항-CD28 항체 (5ug/ml)를 이용하여 1시간동안 활성화시킨 후, 저선량방사선을 10, 25, 50, 100 mGy로 조사하고 24시간 동안 배양 후 세포에서 total RNA를 분리하였다. 이때 저선량방사선을 조사하지 않은 활성화된 CD4+ T 세포를 대조군으로 사용하기위해 total RNA를 분리하였다. 추출된 RNA를 이용하여 miScript II RT Kit (Qiagen, Hilden, Germany)를 이용하여 cDNA를 합성하였으며, 이를 이용하여 정량 PCR을 ABI PRISM 7500 Sequence Detection System (ABI, Foster city, CA) 및 miScript SYBR Green Kit (Qiagen, Hilden, Germany)를 사용하여 발현을 분석하였다. 정량 PCR에 사용된 프라이머의 서열은 표 1과 같으며, 내부 대조군으로 RNU6-2을 사용하였다. In order to confirm the response regulator of miRNAs (mir-142-5p.mir-155-5p) identified in Example 2 in a human cell line, low-dose radiation using the human cutaneous T Lymphocyte cell line HuT78 It was attempted to additionally verify the response regulator by. HuT78 cell line was cultured in RPMI1640 medium supplemented with 20% FBS, 100U/ml penicillin and 100ug/ml streptomycin (Gibco, USA) at 37°C and 5% CO 2 . For low-dose radiation irradiation, 80% of ionizing radiation ( 137 Cs, 930 mGy/min, Grammacell 40 Exactor, Best Theratronics Ltd.) is shielded at a dose rate (186 mGy/min) for each of 4 doses (10, 25, 50, 100). mGy). Cell line HuT78 was activated with anti-CD3 antibody (5ug/ml) and anti-CD28 antibody (5ug/ml) for 1 hour, then irradiated with low-dose radiation at 10, 25, 50, 100 mGy and incubated for 24 hours. After, total RNA was isolated from the cells. At this time, total RNA was isolated to use activated CD4+ T cells that were not irradiated with low-dose radiation as a control. CDNA was synthesized using the miScript II RT Kit (Qiagen, Hilden, Germany) using the extracted RNA, and quantitative PCR was performed using this ABI PRISM 7500 Sequence Detection System (ABI, Foster city, CA) and miScript SYBR Green Kit (Qiagen, Hilden, Germany) was used to analyze the expression. The sequence of the primers used in quantitative PCR is shown in Table 1, and RNU6-2 was used as an internal control.
분석결과는 도 3에 도시하였다. The analysis results are shown in FIG. 3.
도 3을 참고하면, 도 1 및 2에서 확인된 바와 같이, miRNA (mir-142-5p. mir-155-5p)들이 인간 면역세포주에서도 실시예1 및 2와 유사하게 저선량방사선에 반응하여 발현이 변화하는 것을 확인하였다. Referring to Figure 3, as confirmed in Figures 1 and 2, miRNA (mir-142-5p. mir-155-5p) is expressed in response to low-dose radiation in a human immune cell line similar to Examples 1 and 2 It was confirmed to change.
상기 실시예 1 내지 3의 결과는 저선량방사선 피폭에 대하여 특이적 반응 조절인자 mir-142-5p 및 mir-155-5p가 마우스의 면역조직 (비장, 골수)과 인간세포에서 유사하게 조절된다는 것을 확인하였다. 이에 저선량방사선 피폭에 대하여 특이적 반응 조절인자 mir-142-5p 및 mir-155-5p를 저선량방사선 피폭에 대한 선량별 바이오마커로 사용 가능하여 의료 및 방사선 작업 종사자 그리고 일반인의 방사선 피폭을 빠르고 효율적으로 진단할 수 있는 방법 및 선량지표의 진단용 조성물로 제공하고자 한다. The results of Examples 1 to 3 confirmed that specific response modulators mir-142-5p and mir-155-5p were similarly regulated in immune tissues (spleen, bone marrow) of mice and human cells to low-dose radiation exposure. I did. Therefore, specific response regulators mir-142-5p and mir-155-5p for low-dose radiation exposure can be used as dose-specific biomarkers for low-dose radiation exposure, so that radiation exposure of medical and radiation workers and the general public can be quickly and efficiently. It is intended to be provided as a diagnostic method and a composition for diagnosis of a dose index.
이상, 본 발명내용의 특정한 부분을 상세히 기술하였는바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적인 기술은 단지 바람직한 실시양태일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의해 정의된다고 할 것이다. As described above, specific parts of the present invention have been described in detail, and for those of ordinary skill in the art, it is obvious that these specific techniques are only preferred embodiments, and the scope of the present invention is not limited thereby. something to do. Therefore, it will be said that the practical scope of the present invention is defined by the appended claims and their equivalents.
<110> Korea Hydro & Nuclear Power Co. Ltd. <120> A BIOMARKER FOR EACH DOSE OF LOW DOSE RADIATION EXPOSURE, A DIAGNOSTIC METHOD USING THE BIOMARKER, AND A DIAGMOSITIC KIT INCLUDING THE BIOMARKER <130> P19-E053 <160> 4 <170> KoPatentIn 3.0 <210> 1 <211> 21 <212> RNA <213> homo sapiens <400> 1 cataaagtag aaagcactac t 21 <210> 2 <211> 23 <212> RNA <213> homo sapiens <400> 2 ttaatgctaa tcgtgatagg ggt 23 <210> 3 <211> 21 <212> RNA <213> mus musculus <400> 3 cataaagtag aaagcactac t 21 <210> 4 <211> 23 <212> RNA <213> Mus musculus <400> 4 ttaatgctaa ttgtgatagg ggt 23 <110> Korea Hydro & Nuclear Power Co. Ltd. <120> A BIOMARKER FOR EACH DOSE OF LOW DOSE RADIATION EXPOSURE, A DIAGNOSTIC METHOD USING THE BIOMARKER, AND A DIAGMOSITIC KIT INCLUDING THE BIOMARKER <130> P19-E053 <160> 4 <170> KoPatentIn 3.0 <210> 1 <211> 21 <212> RNA <213> homo sapiens <400> 1 cataaagtag aaagcactac t 21 <210> 2 <211> 23 <212> RNA <213> homo sapiens <400> 2 ttaatgctaa tcgtgatagg ggt 23 <210> 3 <211> 21 <212> RNA <213> mus musculus <400> 3 cataaagtag aaagcactac t 21 <210> 4 <211> 23 <212> RNA <213> Mus musculus <400> 4 ttaatgctaa ttgtgatagg ggt 23
Claims (11)
microRNA-142-5p(miR-142-5p) 또는 microRNA-155-5p(miR-155-5p)를 바이오마커로 하여, 상기 바이오마커의 발현 증가 여부로 10 내지 100 mSv 의 저선량방사선 피폭 여부를 진단하는 저선량방사선 피폭 진단 키트.
A diagnostic kit for low-dose radiation exposure using CD4+ T cells in the spleen, bone marrow cells from which red blood cells have been removed, or human T cell lymphoma cell lines as a specimen,
Using microRNA-142-5p (miR-142-5p) or microRNA-155-5p (miR-155-5p) as a biomarker, diagnosis of low-dose radiation exposure of 10 to 100 mSv by increasing the expression of the biomarker Low-dose radiation exposure diagnostic kit.
상기 microRNA-155-5p(miR-155-5p)는 10 내지 100 mSv의 저선량방사선 노출시 10 내지 100 mSv 의 저선량방사선 비노출에 비해 특이적으로 발현이 증가하는 것을 특징으로 하는 저선량방사선 피폭 진단 키트.
The method of claim 1,
The microRNA-155-5p (miR-155-5p) is a low-dose radiation exposure diagnostic kit, characterized in that when exposed to low-dose radiation of 10 to 100 mSv, the expression is specifically increased compared to non-exposed to low-dose radiation of 10 to 100 mSv.
상기 세포의 miRNA(microRNA)를 분석하는 단계; 및
상기 miRNA(microRNA) 중에 microRNA-142-5p(miR-142-5p) 또는 microRNA-155-5p(miR-155-5p)를 10 내지 100 mSv 의 저선량방사선에 비노출된 대조군과 비교하는 단계를 포함하는 저선량방사선 피폭 진단방법.
Preparing bone marrow cells from which CD4+ T cells or red blood cells have been removed from the spleen from animals other than humans as a specimen;
Analyzing the cell miRNA (microRNA); And
Comprising the step of comparing microRNA-142-5p (miR-142-5p) or microRNA-155-5p (miR-155-5p) in the miRNA (microRNA) with a control unexposed to low-dose radiation of 10 to 100 mSv. Low-dose radiation exposure diagnostic method.
상기 miRNA(microRNA)를 분석하는 방법은 정량 RT-PCR(Real-time PCR) 분석 인 것을 특징으로 하는 저선량방사선 피폭 진단방법.
The method of claim 5,
The method of analyzing the miRNA (microRNA) is a diagnostic method for low-dose radiation exposure, characterized in that quantitative RT-PCR (Real-time PCR) analysis.
상기 대조군과 비교하는 단계는 microRNA-142-5p(miR-142-5p) 또는 microRNA-155-5p(miR-155-5p)가 대조군에 비해 특이적으로 발현이 증가되는 것을 확인하는 저선량방사선 피폭 진단방법. The method of claim 5,
The step of comparing with the control group is the diagnosis of low-dose radiation exposure to confirm that the expression of microRNA-142-5p (miR-142-5p) or microRNA-155-5p (miR-155-5p) is specifically increased compared to the control group. Way.
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