Method of detecting radiosensitivity of lung cancer cells on x ray, chip and application

A lung cancer cell, x-ray technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as no relationship between microRNA and tumor radiotherapy sensitivity.

Active Publication Date: 2017-08-29
THE BEIJING PREVENTION & TREATMENT HOSPITAL OF OCCUPATIONAL DISEASE FOR CHEM IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most studies on the relationship between microRNA and tumors focus on the relationship between tumor occurrence, development and prognosis. There are no research reports on the relationship between microRNA and tumor radiotherapy sensitivity at home and abroad.

Method used

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  • Method of detecting radiosensitivity of lung cancer cells on x ray, chip and application
  • Method of detecting radiosensitivity of lung cancer cells on x ray, chip and application
  • Method of detecting radiosensitivity of lung cancer cells on x ray, chip and application

Examples

Experimental program
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Effect test

Embodiment 1

[0064] Conventional molecular biology, microbiology and recombinant DNA techniques within the skill of the art can be used in this example. Such techniques are explained fully in the literature. See, eg, Sambrook, Fritsch & Maniatis, Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press, 1989; DNA Cloning: A Practical Approach, Volumes I and II (ed. D.N. Glover, 1985 ); Oligonucleotide Synthesis (M.J.Gait, ed., 1984); Nucleic Acid Hybridization [B.D.Hames & S.J.Higgins, ed. (1985)]; Transcription and Translation [B.D.Hames & S.J.Higgins, ed. (1984)]; B.Perbal, A Practical Guide to Molecular Cloning (1984); Ausubel, F.M. et al. (eds.); A Current Handbook of Molecular Biology, John Wiley & Sons, Inc., 1994.

[0065] 1. Specimen: select lung cancer cell lines (N-A549) sensitive to radiotherapy (S-A549) and insensitive to radiotherapy, and store them in a -80°C low-temperature refrigerator.

[0066] 2. Total RNA extra...

Embodiment 2

[0110] 1. Construct the overexpression vector of miR-126 (that is, miRNA-126), transfect the SK-MES-1 lung cancer cells, and use the empty load and the mother line as a reference, and use the MTT experiment to detect the changes in the cell proliferation ability after 5Gyx ray irradiation , found that high expression of miR-126 significantly inhibited the proliferation of SK-MES-1 cells (see image 3 ).

[0111] The changes in the radiosensitivity of cells after 0, 2, 4, and 8 Gy x-ray irradiation were detected by colony formation experiments, and it was found that high expression of miR-126 significantly increased the radiosensitivity of SK-MES-1 cells (see Figure 4 ).

[0112] Subsequently, the effect of high expression of miR-126 on radiation-induced apoptosis of SK-MES-1 cells was detected. We found that overexpression of miR-126 could significantly promote the apoptosis of lung cancer cells induced by 4Gy and 8Gy x-ray irradiation (P Figure 5 ).

[0113] The above exp...

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Abstract

The invention discloses a method of detecting radiosensitivity of lung cancer cells on x ray, a chip and an application. The method comprises the following steps: selecting lung cancer cells which are abnormally sensitive to and tolerant to radiotherapy; detecting expression difference of micro RNA in two groups of cells by using a micro RNA expression profile chip; verifying the chip result by means of a Real-Time PCR technology; and finally, screening 11 microRNAs which has remarkable difference. The method disclosed by the invention can be used for predicting sensitivity of the lung cancer cells on radiotherapy.

Description

technical field [0001] The invention relates to a method, chip and application for detecting the sensitivity of lung cancer cells to radiotherapy by using miRNA. Background technique [0002] Lung cancer is one of the most common malignant tumors in my country, and its incidence is increasing year by year. According to the report of the National Cancer Prevention and Research Office, the morbidity and mortality of lung cancer in Beijing, Shanghai, Tianjin, Harbin and other cities are the first among all malignant tumors, accounting for 20% to 30% of all malignant tumors. The 5-year survival rate of surgical treatment of clinical stage Ⅰ and Ⅱ lung cancer is about 40%. Radiation therapy is the main means of lung cancer treatment, about 70% of patients need to receive radiation therapy. In the radiotherapy of lung cancer, the consistent treatment mode for decades is still being applied, 2Gy / time, 5 times a week, with a total dose of 60-70Gy / 6-7 weeks. In fact, different ind...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/106C12Q2600/158C12Q2600/178
Inventor 王小春李珏丁晓文方芳盛文涛
Owner THE BEIJING PREVENTION & TREATMENT HOSPITAL OF OCCUPATIONAL DISEASE FOR CHEM IND
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