CN109825568B - Radiation sensitive gene marker and application thereof in identifying low LET (low-LET-rate) ray radiation - Google Patents
Radiation sensitive gene marker and application thereof in identifying low LET (low-LET-rate) ray radiation Download PDFInfo
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Abstract
The invention provides a radiation-sensitive gene marker and application thereof in identifying low LET (Let radiation). The radiation-sensitive gene markers comprise Wnt7b, Tprkb, Pira1, Pde4dip, Limk2, Ctns, Kcnk6, Csf2rb, Cd80 and Sesn 2. Based on detecting the gene expression level of a radiation-sensitive gene marker in a radiated organism, the method is used for identifying whether the radiated organism is exposed to low LET radiation (such as X-ray, beta-ray, gamma-ray and the like) so as to provide a radiation dose reference for the evaluation of the nuclear radiation damage risk.
Description
Technical Field
The invention belongs to the field of radiation biological ray type detection, and particularly relates to a radiation sensitive gene marker and application thereof in identification of low LET ray radiation.
Background
X rays and other low Linear Energy Transfer (LET) rays such as beta rays and gamma rays, electronic wires and the like have wide application bases in various fields of medical treatment and health, industrial and agricultural production, special medicine, scientific research and the like. LET is a parameter for evaluating radiation quality. Compared with high LET rays (such as heavy ions, fast neutrons, carbon, nitrogen, oxygen, neon and the like), the low LET rays have obviously lower ionization density per unit length, and the scattering effect of the rays is very obvious. In addition to the penetration effect and the fluorescence effect which are inherent to electromagnetic waves, X-ray radiation protection is also a problem which is becoming more and more interesting.
In the daily CT examination and tumor radiotherapy which are used in large quantities, particularly in orthopedics, tumor intervention and cardiac conduction beam ablation operations based on digital X-ray photography or digital subtraction angiography, the radiation protection tools of medical personnel, including lead clothes, neckerchief, eyeshades and the like, can not completely block X-rays in the operations. Causing the medical care personnel in related and adjacent departments (such as operating rooms on upper and lower floors) to have obviously raised malignant tumors such as thyroid cancer, breast cancer and the like. In addition to the application of X-rays in medicine, low LET rays are currently widely used in the fields of industrial exploration, weapon armor and metal quality detection, anti-terrorism security inspection, and the like. The health impact of short-term high-dose or long-term low-dose low-LET ray accumulation on practitioners and examination subjects has become a problem of widespread concern today. The low LET line can cause irreversible damage to organisms and possibly cause adverse reactions such as malignant tumors, blood system diseases, inflammation fibrosis and the like.
In addition, low LET radiation is also one of the types of radiation (e.g., beta particles, gamma rays, etc.) that are released during the explosion of nuclear weapons. The electromagnetic weapons protection faced in modern war also involves various low LET rays. Potential radiation threats may also come from industrial or medical sources that may cause low LET radiation in the event of a breach in the containment of the source or loss of the source during transport, storage, or operation. Radiation sanitation and health monitoring for practitioners are oriented, various radiation emergencies are effectively dealt with, and effective distinguishing and judging of exposed rays based on biological effects are increasingly paid more and more attention. The method has key significance for nuclear safety, treatment of radiation accidents and timely and proper treatment of exposed people.
Disclosure of Invention
Based on the problems of the prior art, the first objective of the present invention is to provide a radiation-sensitive genetic marker which is proved by the applicant to have high specific radiation sensitivity only for low-Linear Energy Transfer (LET) rays (such as X-rays) and no radiation sensitivity for high-Linear Energy Transfer (LET) rays (such as heavy ion rays).
The second purpose of the invention is to provide the application of the radiation-sensitive gene marker in identifying low LET radiation. The radiation dose reference is provided based on detecting the gene expression level of a radiation-sensitive gene marker in the irradiated organism, and is used for identifying whether the irradiated organism is exposed to low LET radiation (such as X-ray, beta-ray, gamma-ray and the like).
The third purpose of the invention is to provide a kit for identifying whether an organism is irradiated by low LET rays and application thereof, and the kit containing the radiation sensitive gene marker can be used for identifying the low LET rays more quickly, conveniently and accurately and providing a dosage reference.
The fourth object of the present invention is to provide a method for obtaining the radiation-sensitive gene marker of the present invention.
The purpose of the invention is realized by the following technical means:
in a first aspect, the present invention provides a radiation-sensitive gene marker comprising 10 radiation-sensitive genes: wnt7B (Wnt Family Member7B, Wnt Family Protein 7B), Tprkb (TP53-Related Protein Kinase Binding Protein ), Pira1(Paired-Ig-Like Receptor A1, Paired-Ig-Like Receptor A1), Pde4dip (Phosphodiesterase 4 DINTracting Protein, Phosphodiesterase 4D interacting Protein), Limk2(LIM Domain Kinase 2), lysosomes 865ns (Cystinon, lysomal Cyctine Transporter, Cystine Transporter), Kcnk6 (PotasCd Two-way Domain Receptor K466, Potassium diplomane Domain Subfamily K6), Sttif 2 (Sttins 2) 4 (Molecular Receptor 5, Molecular Receptor 80), lysosome Cd 2, Molecular Receptor Binding Protein, Colony Stimulating Factor 3625 (Colony 80).
In the above-mentioned radiation-sensitive gene marker, preferably, the nucleotide sequence of Wnt7b is as follows:
AGAGAGGTGGTTAGTGGACCCAGGCAGGGCACTGGCTGTCCCAATGCTGT (shown in SEQ ID NO: 1) (5 '-3');
the nucleotide sequence of Tprkb is as follows:
GCGCCAAGTCTGCAAAGCCAGGTGCTCTCATAGTGCAGTTCTGGGGTTGT (shown in SEQ ID NO: 2) (5 '-3');
the nucleotide sequence of Pira1 is as follows:
CCAGGATCTGTGATCGCCTCCAAAAGAGCAATGACCATCTGGTGTCAGGG (shown in SEQ ID NO: 3) (5 '-3');
the nucleotide sequence of the Pde4dip is as follows:
GGGGGGAAGGAACTAATGACATCGTCTCAGACGTTCATCTCTAACCAGCC (shown in SEQ ID NO: 4) (5 '-3');
the nucleotide sequence of Limk2 is as follows:
TGAGTATGCTTGCACTGTCCCCAGCAAGTGTGGGAGTGGGGCCTGCACTA (shown in SEQ ID NO: 5) (5 '-3');
the nucleotide sequence of Ctns is as follows:
GCCTTCAGAACCAAGTCCTGGGGGCTTAGAGGACCTTGCTTACCTATGTC (shown in SEQ ID NO: 6) (5 '-3');
the nucleotide sequence of the Kcnk6 is as follows:
CAGAGCCCAAGCCACATCTACTACTGTGTGCCTAGCACAGAAAAGCATGG (SEQ ID NO: 7) (5 '-3');
the nucleotide sequence of the Csf2rb is as follows:
TGAGCACACATTCCAGGTCCAGTACAAGAAGAAATCGGACAGCTGGGAGG (shown in SEQ ID NO: 8) (5 '-3');
the nucleotide sequence of Cd80 is as follows:
GCTCTTTGGGGCAGGATTCGGCGCAGTAATAACAGTCGTCGTCATCGTTG (shown in SEQ ID NO: 9) (5 '-3');
the nucleotide sequence of the Sesn2 is as follows:
TGGCTGCCTGTGTGGGAGAGGAGTAAGGACCTCCAGGGACTAGCACTCCA (shown in SEQ ID NO: 10) (5 '-3').
In a second aspect, the present invention also provides the use of a radiation-sensitive gene marker as described above for the identification of low LET radiation.
In the above application, preferably, the application comprises the following steps:
detecting the gene expression levels of radiation-sensitive gene markers Wnt7b, Tprkb, Pira1, Pde4dip, Limk2, Ctns, Kcnk6, Csf2rb, Cd80 and Sesn2 in the irradiated organism, and identifying the irradiated organism as low LET ray radiation exposure when the expression level of the genes is obviously high-regulated.
In the above applications, the organism may include a human, an animal, and the like.
In the above application, preferably, the low LET radiation includes one or more of X-ray, β -ray, γ -ray, and the like.
In the above application, preferably, the gene expression level of the sensitive gene marker is an average value of the transcription level difference expression fold.
In a third aspect, the present invention also provides a kit for identifying whether an organism is irradiated by low LET radiation, the kit comprising the above-described radiation-sensitive gene marker.
In a fourth aspect, the invention also provides the use of a kit as described above for the identification of low LET radiation.
In a fifth aspect, the present invention also provides a method for obtaining the above-mentioned radiation-sensitive gene marker for non-therapeutic purposes, comprising the steps of:
after the mice are irradiated by single gradient doses and graded gradient doses with different ray types and different doses, fresh lung tissues are extracted to carry out gene chip or real-time quantitative polymerase chain reaction, the whole genome expression level of the 24-week lung tissues is analyzed on the basis of a whole genome gene expression array, the trend relation between different ray irradiation doses and gene expression is analyzed, and an expression gene combination which is obtained, wherein the gene expression level is increased along with the increase of the low LET ray irradiation dose and is not changed along with the change of the high LET ray irradiation dose, namely the radiation sensitive gene marker, is obtained.
In the above method, the single gradient dose irradiation and the fractionated gradient dose irradiation with different radiation types and different doses may be performed in the following manners, for example: x-ray gradient dose fraction irradiation (0, 10.5, 12.5, 14.5, 17.5Gy), X-ray gradient dose fraction irradiation (5 irradiation in 0, 10, 20, 30, 40 Gy), Carbon ion (Carbon-ions) gradient dose fraction irradiation (0, 7.5, 10.5, 12.5, 14.5Gy), Carbon ion (Carbon-ions) gradient dose fraction irradiation (5 irradiation in 0, 5, 10, 15, 20 Gy).
In the above method, preferably, the different ray types include a low LET ray and a high LET ray; more preferably, the high LET rays include one or more of fast neutron rays, negative pi meson rays, heavy ion rays and the like.
The invention relates to a biological dose evaluation means established according to the stress change of the transcription level of relevant radiation sensitive genes of mouse lung tissues after X-ray or other low LET rays are irradiated.
The invention screens out related sensitive gene combinations by utilizing whole genome transcriptomics analysis 24 weeks after mice receive different LET rays and are irradiated on the whole lung. The specific gene combination has no response to high LET rays, but can be driven to generate obvious high regulation after being irradiated by low LET rays, and the X-ray radiation identification is quickly carried out according to a gene expression level-X-ray prediction model through the biological effect.
The mouse model adopted by the invention is a mouse radioactive lung injury model, is an important tool for researching the relation between radiation pneumonitis, pulmonary fibrosis and radiation dose, normal tissue radiation reaction, respiratory system diseases and immune response reaction mechanism, and has irreplaceable effects in radiobiology, radiotherapy and research and development of lung injury resistant drugs. The early stages of radiation lung injury are primarily manifested as radiation pneumonitis (usually secondary to within 3 months after irradiation), a lymphocytic alveolitis, due to the immune response generated by the activation of a large number of T lymphocytes; advanced stage can develop into interstitial pulmonary fibrosis (about 6 months after irradiation). The pathophysiological process of the radioactive lung injury of the mouse is very similar to that of the human, so that the radioactive lung injury of the mouse can be used as a reliable reference experiment model.
In the invention, if the organism receives mixed rays, namely, the mixed rays contain high LET rays and low LET rays, the method can accurately identify the low LET ray part.
The invention has the beneficial effects that:
(1) the radiation-sensitive gene marker has high specific radiation sensitivity only aiming at low-Linear Energy Transfer (LET) rays (such as X rays and the like), has no radiation sensitivity to high-Linear Energy Transfer (LET) rays (such as heavy ion rays and the like), and is used for identifying whether a radiated organism is exposed to low LET rays (such as X rays, beta rays, gamma rays and the like) based on detecting the gene expression level of the radiation-sensitive gene marker in the radiated organism, thereby providing a radiation dose reference for the risk evaluation of nuclear radiation damage.
(2) The kit containing the radiation-sensitive gene marker can be used for identifying low LET rays more quickly, conveniently and accurately and providing a dosage reference.
Drawings
FIG. 1 is a graph of Venn diagram of the subset of dose-dependent up-regulated genes (dose-dependent up-regulation genes) [ X-ray gradient dose single irradiation (0, 10.5, 12.5, 14.5, 17.5Gy), X-ray gradient dose fraction irradiation (0, 10, 20, 30, 40Gy divided by 5 irradiation), Carbon ion (Carbon-ions) gradient dose single irradiation (0, 7.5, 10.5, 12.5, 14.5Gy), Carbon ion (Carbon-ions) gradient dose fraction irradiation (0, 5, 10, 15, 20Gy divided by 5 irradiation) ], obtained after 24 weeks of analysis of the whole genome transcription level of lung tissue in example 1 of the present invention in C57BL/6 mice.
FIG. 2 is a chart showing the heatmap of 126X-ray specific genes obtained by analysis of a Venn diagram in example 1 of the present invention.
FIG. 3 is a gene heatmap of low LET radiation biology identification gene combinations for the most significant (Top 10) X-ray specific gene composition of the Venn diagram analysis in example 1 of the present invention.
FIG. 4 shows the expression levels (log2 fold-change) of 10 specific sensitive genes obtained by screening in example 2 of the present invention 24 weeks after X-ray irradiation.
FIG. 5 is the gene expression values (P <0.0001) of 10 specific sensitive genes screened in example 2 of the present invention at 24 weeks after gradient dose X-ray and heavy ion irradiation.
FIG. 6 is a diagram of the receiver operating characteristic curve (ROC).
Detailed Description
The technical solutions of the present invention will be described in detail below in order to clearly understand the technical features, objects, and advantages of the present invention, but the present invention is not limited to the practical scope of the present invention.
The chemical reagents and the like used in the following examples are commercially available unless otherwise specified.
Example 1 screening acquisition of radiation sensitive Gene markers
1. Animal experiment irradiation and grouping: c57BL6 female mice 8-10 weeks old are bred in SPF grade under the conditions of temperature (23 +/-1) DEG C, relative humidity 55 +/-5%, pressure difference less than or equal to 10Pa, light irradiation for 12h every day, and free drinking water.
2. After isoflurane induced anesthesia is carried out before irradiation, the mouse breast is transferred to a special mouse breast irradiation device and is properly fixed. Irradiation of the whole lung field was performed using X-rays and heavy ion (carbon-ions) rays. X-rays are representative of low LET rays and are irradiated after accelerated by a medical linear accelerator (energy 6-10 MV). Heavy ion rays, which are the main representatives of high LET rays, are carbon particle beams (energy 280MeV) which are drained after being accelerated by a cyclotron (the diameter reaches 20-30 meters), and have extremely high energy and speed.
3. The irradiation and grouping includes X-ray gradient dose single irradiation: 0. 10.5, 12.5, 14.5, 17.5Gy, X-ray gradient dose fractionated irradiation: 0. 10, 20, 30 and 40Gy are divided into 5 times of irradiation and single irradiation with heavy ion gradient dose: 0. 7.5, 10.5, 12.5, 14.5Gy, heavy ion gradient dose fractionated irradiation: 0.5, 10, 15 and 20Gy irradiation times were divided into 5.
4. The blank control group received no radiation (0 Gy). Control and dose groups were randomly assigned 3 mice per group.
5. Fresh lung tissue was taken 24 weeks after irradiation, treated with total RNA extraction reagent (Trizol), and total RNA was isolated and extracted using RNeasy Mini kit. To avoid DNA contamination, RNA was treated with DNase I. Purified RNA was eluted in 45.0. mu.l nuclease-free water and stored at-20.0 ℃. RNA concentration and purity were assessed using a NanoDrop ND-1000 spectrophotometer. The integrity and purity of the RNA samples were determined using a 2100Bioanalyzer and the corresponding RNA Nano Chip. Samples were analyzed based on whole genome gene expression arrays. And generating an expression matrix, annotating data and processing by using a self-contained software package. In subsequent statistical tests, the data were log2 transformed to account for increased or decreased gene expression. In order to analyze the relation between radiation and gene expression trend at different time points, R language software package is adopted for carrying out cluster analysis, including principal component analysis, hierarchical cluster, K mean cluster and self-organizing mapping to identify the gene expression related to the dosage.
Analyzing the whole genome expression level of 24-week lung tissues based on a whole genome gene expression array, analyzing the trend relation between different radiation doses and gene expression under the irradiation of rays, and obtaining an expression gene combination which is the radiation sensitive gene marker and the gene expression level of which is increased along with the increase of the radiation dose of low LET rays and is not changed along with the change of the radiation dose of high LET rays.
The specific screening process is as follows:
referring to fig. 1 and 2, a subset of dose-dependent up-regulated genes (dose-dependent up-regulation genes) was obtained after analysis of the whole genome transcript level of lung tissue 24 weeks after X-ray gradient dose bolus irradiation (0, 10.5, 12.5, 14.5, 17.5Gy), X-ray gradient dose fractionated irradiation (0, 10, 20, 30, 40Gy divided by 5 irradiations), Carbon ion (Carbon-ions) gradient dose bolus irradiation (0, 5, 10, 15, 20Gy divided by 5 irradiations). Analysis of the Venn diagram yielded 126X-ray specific genes.
Referring to fig. 3, the heat map analysis of the lung tissue gene expression levels of the mice after different radiation irradiation respectively shows that: the gene expression level is increased along with the increase of X-ray dose, and is in dose correlation; the gene expression level is not changed with the change of heavy ion radiation dose. 10 radiation-sensitive gene combinations were obtained by screening: wnt7b (Wnt Family Member7B, Wnt Family Protein 7b), Tprkb (TP53-Related Protein Kinase Binding Protein ), Pira1(Paired-Ig-Like Receptor A1, Paired-Ig-Like Receptor A1), Pde4dip (Phosphodiesterase 4D Interacting Protein ), Limk2(LIM Domain Kinase 2, LIM Domain Kinase 2), lysosomes (Ctstinsin, Lysosomal cysteine Transporter, Cystine Transporter), Kcnk6 (Potassyl Two Domain scaffold K Receptor K6, Potassium diplopore Domain Channel Family Subfamily K6), Cstmill 2rb (Protein Kinase 2 Cd 2), Molecular gene 7372, Cd 2Receptor Subunit 2, Colony Stimulating Factor Cd 2, 2Receptor 80, and Syngnac 2Receptor molecules (Protein).
In the above-mentioned radiation-sensitive gene marker, preferably, the nucleotide sequence of Wnt7b is as follows:
AGAGAGGTGGTTAGTGGACCCAGGCAGGGCACTGGCTGTCCCAATGCTGT (shown in SEQ ID NO: 1) (5 '-3');
the nucleotide sequence of Tprkb is as follows:
GCGCCAAGTCTGCAAAGCCAGGTGCTCTCATAGTGCAGTTCTGGGGTTGT (shown in SEQ ID NO: 2) (5 '-3');
the nucleotide sequence of Pira1 is as follows:
CCAGGATCTGTGATCGCCTCCAAAAGAGCAATGACCATCTGGTGTCAGGG (shown in SEQ ID NO: 3) (5 '-3');
the nucleotide sequence of the Pde4dip is as follows:
GGGGGGAAGGAACTAATGACATCGTCTCAGACGTTCATCTCTAACCAGCC (shown in SEQ ID NO: 4) (5 '-3');
the nucleotide sequence of Limk2 is as follows:
TGAGTATGCTTGCACTGTCCCCAGCAAGTGTGGGAGTGGGGCCTGCACTA (shown in SEQ ID NO: 5) (5 '-3');
the nucleotide sequence of Ctns is as follows:
GCCTTCAGAACCAAGTCCTGGGGGCTTAGAGGACCTTGCTTACCTATGTC (shown in SEQ ID NO: 6) (5 '-3');
the nucleotide sequence of the Kcnk6 is as follows:
CAGAGCCCAAGCCACATCTACTACTGTGTGCCTAGCACAGAAAAGCATGG (SEQ ID NO: 7) (5 '-3');
the nucleotide sequence of the Csf2rb is as follows:
TGAGCACACATTCCAGGTCCAGTACAAGAAGAAATCGGACAGCTGGGAGG (shown in SEQ ID NO: 8) (5 '-3');
the nucleotide sequence of Cd80 is as follows:
GCTCTTTGGGGCAGGATTCGGCGCAGTAATAACAGTCGTCGTCATCGTTG (SEQ ID NO: 9) (5 '-3');
the nucleotide sequence of the Sesn2 is as follows:
TGGCTGCCTGTGTGGGAGAGGAGTAAGGACCTCCAGGGACTAGCACTCCA (shown in SEQ ID NO: 10) (5 '-3').
Example 2
This example provides the use of the radiation sensitive gene markers obtained by screening in example 1 for the identification of low LET radiation.
Referring to fig. 4, the expression fold difference of 10 genes after the gradient dose X-ray and carbon ion ray irradiation, the heavy ion ray irradiation group had almost no radiation response.
Referring to fig. 5, the gene expression values of 10 genes were different in 24 weeks of gradient X-ray and carbon ion irradiation, and the difference was statistically significant (P < 0.0001).
Referring to fig. 6, a receiver operating characteristic curve (ROC) graph shows the discrimination ability of the average gene expression value of 10 genes to X-ray after irradiation of gradient dose X-ray and carbon ion ray. The Area under the Curve (Area under the Curve, AUC) was equal to 0.984, the 95% Confidence Interval (CI) was 0.952-1.000, and when the average of the above 10 gene expression values was equal to 1.033, the sensitivity to X-rays was identified to be 95.83%, the specificity was 100%, indicating that the above 10 gene expression values had good discriminative ability for X-ray radiation.
The radiation sensitive gene marker is made into a kit for identifying whether the low LET rays are radiated or not, so that the low LET rays can be identified more quickly, conveniently and accurately, the dosage reference is provided, and the kit is favorable for marketing and using.
The above-mentioned embodiments are only for illustrating the technical ideas and features of the present invention, and the purpose thereof is to enable those skilled in the art to understand the contents of the present invention and to implement the same, and the present invention shall not be limited to the embodiments, i.e. the spirit of the present invention disclosed, and equivalent changes or modifications shall still fall within the scope of the present invention.
Sequence listing
<110> military medical research institute of military science institute of people's liberation force of China
<120> radiation-sensitive gene marker and application thereof in identification of low LET (low-LET-rate) radiation
<130> GAI18CN6324
<160> 10
<170> PatentIn version 3.5
<210> 1
<211> 50
<212> DNA
<213> Wnt7b
<400> 1
agagaggtgg ttagtggacc caggcagggc actggctgtc ccaatgctgt 50
<210> 2
<211> 50
<212> DNA
<213> Tprkb
<400> 2
gcgccaagtc tgcaaagcca ggtgctctca tagtgcagtt ctggggttgt 50
<210> 3
<211> 50
<212> DNA
<213> Pira1
<400> 3
ccaggatctg tgatcgcctc caaaagagca atgaccatct ggtgtcaggg 50
<210> 4
<211> 50
<212> DNA
<213> Pde4dip
<400> 4
ggggggaagg aactaatgac atcgtctcag acgttcatct ctaaccagcc 50
<210> 5
<211> 50
<212> DNA
<213> Limk2
<400> 5
tgagtatgct tgcactgtcc ccagcaagtg tgggagtggg gcctgcacta 50
<210> 6
<211> 50
<212> DNA
<213> Ctns
<400> 6
gccttcagaa ccaagtcctg ggggcttaga ggaccttgct tacctatgtc 50
<210> 7
<211> 50
<212> DNA
<213> Kcnk6
<400> 7
cagagcccaa gccacatcta ctactgtgtg cctagcacag aaaagcatgg 50
<210> 8
<211> 50
<212> DNA
<213> Csf2rb
<400> 8
tgagcacaca ttccaggtcc agtacaagaa gaaatcggac agctgggagg 50
<210> 9
<211> 50
<212> DNA
<213> Cd80
<400> 9
gctctttggg gcaggattcg gcgcagtaat aacagtcgtc gtcatcgttg 50
<210> 10
<211> 50
<212> DNA
<213> Sesn2
<400> 10
tggctgcctg tgtgggagag gagtaaggac ctccagggac tagcactcca 50
Claims (8)
1. A radiation-sensitive gene marker combination characterized by: the radiation-sensitive gene marker combination consists of Wnt7b, Tprkb, Pira1, Pde4dip, Limk2, Ctns, Kcnk6, Csf2rb, Cd80 and Sesn2, wherein the radiation is X-ray radiation, and the radiation-sensitive gene marker is an expression gene of which the gene expression quantity is increased along with the increase of the X-ray radiation dose and is not changed along with the change of the carbon ion radiation dose.
2. The radiation-sensitive gene marker combination of claim 1, wherein:
the nucleotide sequence of Wnt7b is shown in SEQ ID NO: 1 is shown in the specification;
the nucleotide sequence of the Tprkb is shown as SEQ ID NO: 2 is shown in the specification;
the nucleotide sequence of Pira1 is shown as SEQ ID NO: 3 is shown in the figure;
the nucleotide sequence of the Pde4dip is shown as SEQ ID NO: 4 is shown in the specification;
the nucleotide sequence of the Limk2 is shown as SEQ ID NO: 5 is shown in the specification;
the nucleotide sequence of the Ctns is shown as SEQ ID NO: 6 is shown in the specification;
the nucleotide sequence of the Kcnk6 is shown as SEQ ID NO: 7 is shown in the specification;
the nucleotide sequence of the Csf2rb is shown as SEQ ID NO: 8 is shown in the specification;
the nucleotide sequence of the Cd80 is shown as SEQ ID NO: 9 is shown in the figure;
the nucleotide sequence of the Sesn2 is shown as SEQ ID NO: shown at 10.
3. Use of a reagent for detecting the gene expression level of the radiation-sensitive gene marker combination of claim 1 or 2 in the preparation of a kit for identifying X-ray radiation exposure and carbon ion radiation exposure.
4. Use according to claim 3, characterized in that it comprises the following steps:
detecting the gene expression levels of radiation-sensitive gene markers Wnt7b, Tprkb, Pira1, Pde4dip, Limk2, Ctns, Kcnk6, Csf2rb, Cd80 and Sesn2 in the irradiated organism, and determining that the irradiated organism is exposed to X-ray radiation when the expression level of the genes is remarkably high, wherein the organism is a mouse.
5. Use according to claim 3, characterized in that: the gene expression level of the sensitive gene marker is the average value of the transcription level difference expression fold.
6. A kit for identifying X-ray radiation exposure from carbon ionizing radiation exposure, comprising: the kit contains a reagent for detecting the gene expression level of the radiation-sensitive gene marker combination according to claim 1 or 2 in an organism irradiated, wherein the organism is a mouse.
7. Use of a kit according to claim 6 for the manufacture of a kit for discriminating between X-ray radiation exposure and carbon ion radiation exposure.
8. A method for obtaining the radiation sensitive gene marker combination of claim 1 or 2 for non-therapeutic purposes, comprising the steps of:
after mice are irradiated with different ray types, different doses of single gradient doses and graded gradient doses of whole lung fields, extracting fresh lung tissues to perform gene chip or real-time quantitative polymerase chain reaction, analyzing the whole genome expression level of 24-week lung tissues based on a whole genome gene expression array, analyzing the trend relation of different ray irradiation, radiation dose and gene expression, and obtaining an expression gene combination, namely a radiation sensitive gene marker combination, of which the gene expression quantity is increased along with the increase of the X-ray radiation dose and is not changed along with the change of the carbon ion ray radiation dose;
wherein the different ray types, different doses of single gradient dose irradiation and fractionated gradient dose irradiation are: the X-ray gradient dose irradiation is 0, 10.5, 12.5, 14.5 and 17.5Gy, the X-ray gradient dose irradiation is 0, 10, 20, 30 and 40Gy for 5 times, the carbon ion ray gradient dose irradiation is 0, 7.5, 10.5, 12.5 and 14.5Gy, and the carbon ion ray gradient dose irradiation is 0, 5, 10, 15 and 20Gy for 5 times.
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