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Application of Radiation Sensitive Gene as Radiation Biological Dosimeter

A radiation biological dose and radiation sensitive technology, applied in the field of radiation biological dosimetry, can solve the problems of low accuracy, high analysis technology requirements, long time-consuming, etc., and achieve the effect of high accuracy

Active Publication Date: 2022-07-12
NAT INST FOR RADIOLOGICAL PROTECTION & NUCLEAR SAFETY CHINESE CENT FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to solve the above problems, the present invention preliminarily screens out hundreds of radiation-responsive genes by detecting the changes in the expression of genes contained in the peripheral blood of normal people after being subjected to ionizing radiation; furthermore, the present invention obtains Further research was carried out on genes that showed a good dose-response relationship after exposure to ionizing radiation and could be plotted for R 2 Higher (R of the fitted curve 2 13 radiosensitive genes (PCNA, CCNG1, CDKN1A, XPC, TNFSF4, DDB2, ASTN2, PHPT1, GDF15, TNFRSF10B, BBC3, BAX and FDXR) of the dose-response curve whose value is more than 0.6); Based on the above-mentioned 13 radiation-sensitive genes, the combination of radiation-sensitive genes used as a radiation biological dosimeter to estimate radiation dose was determined, and the accuracy of estimating radiation dose was confirmed by in vivo and in vitro experiments. In the dose estimation method, there are problems such as high requirements for analysis technology, long time consumption, and low accuracy, thus completing the present invention

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  • Application of Radiation Sensitive Gene as Radiation Biological Dosimeter
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  • Application of Radiation Sensitive Gene as Radiation Biological Dosimeter

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Preliminary screening of radiation-sensitive genes

[0032] The gene chip technology was used to detect the changes in the expression of genes contained in normal human peripheral blood after being subjected to ionizing radiation, and hundreds of genes with radiation response were preliminarily screened. Furthermore, the genes obtained from the above preliminary screening were further studied, and a good dose-response relationship was obtained after being exposed to ionizing radiation, and R was drawn. 2 Thirteen radiation-sensitive genes (ie, PCNA, CCNG1, CDKN1A, XPC, TNFSF4, DDB2, ASTN2, PHPT1, GDF15, TNFRSF10B, BBC3, BAX, and FDXR) with higher dose-response curves.

[0033] The details of the studies on these 13 radiation-sensitive genes in the above preliminary screening are as follows:

[0034] 1. Primer Design and Synthesis

[0035] According to the gene information in the GenBank database, 13 radiation-sensitive genes (PCNA, CCNG1, CDKN1A, XPC, TNFSF...

Embodiment 2

[0048] Example 2: Radiation Sensitive Gene Combination as Radiation Biological Dosimeter for Estimating Radiation Dose

[0049] 1. Primer Design and Synthesis

[0050] The same primers as in Example 1 were used for the 13 radiation-sensitive genes and the 2 reference genes (shown in Table 1).

[0051] 2. The blood sample collection and irradiation were performed in the same manner as in Example 1.

[0052] 3. Cell total RNA extraction and cDNA synthesis were performed in the same manner as in Example 1.

[0053] 4. Real-time quantitative PCR detection was performed in the same manner as in Example 1.

[0054] 5. The PCR data processing was carried out in the same manner as in Example 1.

[0055] 6. Establishment of Radiation Sensitive Gene Portfolio

[0056] The multivariate stepwise regression method is used to establish the radiation-sensitive gene combination. The basic principle is: according to the effect of the independent variable on the dependent variable, the regres...

Embodiment 3

[0066] Example 3: Validation of radiation-sensitive gene combinations in double-blind in vitro experiments

[0067] 1. Blood sample collection and irradiation

[0068] On the premise of signing the informed consent, peripheral blood was collected from 10 healthy people, half male and female, 8 ml of venous blood samples were collected from each person, and aliquoted into two 4 ml lithium heparin anticoagulation vacuum blood collection tubes for use at room temperature. 60 Coγ irradiation (double-blind method), the dose rate was 1 Gy / min, and the dose points were 0.8, 1.5, 2.5, 3.5, 4.5 and 5 Gy, respectively. After exposure, 2 ml of blood samples at each dose were inoculated into RPMI 1640 culture flasks containing 8 ml of 10% fetal bovine serum, and placed in a constant temperature incubator at 37 °C for 24 hours; the other 2 ml of whole blood were added to two flasks containing 4 ml of RPMI 1640. (containing 20% ​​fetal bovine serum, 100 U / ml penicillin, 100 μg / ml streptomy...

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Abstract

The invention relates to the application of a combination of multiple radiation sensitive genes as a radiation biological dosimeter in radiation dose estimation. Specifically, the present invention relates to the application of a combination of 5-13 different radiation-sensitive genes as a radiation biological dosimeter in radiation dose estimation, including preparing a combination of 5-13 different radiation-sensitive genes for radiation dose estimation based on the combination of the 5-13 different radiation-sensitive genes A kit for estimating dose, a microarray, and a method for estimating radiation dose using the kit and microarray. Compared with traditional methods including chromosomal aberration analysis, the radiation biological dosimeter and application method have the advantages of simplicity, rapidity, high throughput, etc., and the accuracy of dose estimation is high, especially suitable for nuclear accidents and radiation accidents. Radiation dose estimation for a range population sample.

Description

technical field [0001] The invention belongs to the field of radiation biological dosimetry, and relates to the application of a combination of multiple radiation sensitive genes as a radiation biological dosimeter in radiation dose estimation. Specifically, the present invention relates to the application of a combination of 5-13 different radiation-sensitive genes as a radiation biological dosimeter in radiation dose estimation, including preparing a combination of 5-13 different radiation-sensitive genes for radiation dose estimation based on the combination of the 5-13 different radiation-sensitive genes A kit for estimating dose, a microarray, and a method and application for estimating radiation dose. Background technique [0002] With the wide application of nuclear energy and ionizing radiation, nuclear accidents and radiation accidents occur from time to time. In the early stage of the accident, many people may be exposed. In this case, how to judge a large range of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/166C12Q2600/158
Inventor 刘青杰李爽陆雪封江彬田梅
Owner NAT INST FOR RADIOLOGICAL PROTECTION & NUCLEAR SAFETY CHINESE CENT FOR DISEASE CONTROL & PREVENTION
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