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Method for screening nasopharyngeal carcinoma radiotherapy resistant cells by using radioactive irradiation mode

A technology of radiotherapy and irradiation mode for nasopharyngeal carcinoma, which is applied in the field of screening nasopharyngeal carcinoma radiotherapy-resistant cells by radiation irradiation mode, which can solve the problems of difficulty in establishing radiotherapy resistance, difficulty in surviving cells, and increased chance of contamination, so as to increase the generation of radiotherapy resistance Chance, shortened recovery time, shortened growth recovery time

Inactive Publication Date: 2014-09-03
XIANGYA HOSPITAL CENT SOUTH UNIV
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Problems solved by technology

[0002]At present, most of the construction methods of radiotherapy-resistant cells of nasopharyngeal carcinoma at home and abroad adopt the mode of sublethal dose (10-12 Gy) to the parental cells of the tumor (such as "Cancer "Identification of biomarkers for predicting nasopharyngeal carcinoma response to radiotherapy by proteomics" published in Volume 70 of Issue 9 of Res in 2010;) or small-dose fractionated irradiation mode (1.8-2 Gy) (such as "Radiat Res" 2001 No. Volume 156 of Issue 6 published "The generation and characterization of a radiation-resistant model system to study radioresistance in human breast cancer cells "A paper) for screening methods, but the sublethal dose screening mode has cells that are difficult to survive and prone to degeneration Deficiencies such as death and long screening period; low-dose fractionated irradiation method has problems such as high number of irradiations, difficulty in establishing radiotherapy resistance, and increased chance of contamination

Method used

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Embodiment 1

[0016] This embodiment includes the following steps:

[0017] (1) Dilute CNE-2 nasopharyngeal carcinoma cells in exponential growth phase to about 1000 cells / ml after digested with trypsin. Cells at this concentration can disperse the cells evenly, prevent them from forming clusters, and easily form colonies;

[0018] (2) Add the diluted cells into the culture flask and place at 37°C, 5% CO 2 , saturated humidity, cultivated under sterile conditions for 6 hours, so that the cells are in a state of adherence but no proliferation or a small amount of proliferation;

[0019] (3) Radiation exposure (radiotherapy rays are X-rays): first time: 2Gy, second time: 2Gy, third time: 4Gy, fourth time: 4Gy, fifth time: 6Gy, sixth time: 6Gy, The seventh time: 8Gy, the eighth time: 8Gy, the ninth time: 10Gy, the tenth time: 10Gy, the eleventh time: 10Gy; the total radiotherapy dose is 70Gy; the cells formed after radiotherapy are named as radiotherapy resistant cells.

[0020] (4) After ea...

Embodiment 2

[0023] This embodiment includes the following steps:

[0024] (1) Dilute the 6-10B nasopharyngeal carcinoma cell line in the exponential growth phase to 500 cells / ml after digested with trypsin. The cells at this concentration can make the cells disperse evenly and prevent them from agglomerating, which is easy to form colonies;

[0025] (2) After dilution, the cells are added to the culture dish and placed at 37°C, 5% CO 2 , saturated humidity, cultivated under sterile conditions for 8 hours, so that the cells are in a state of adherence but no proliferation or a small amount of proliferation;

[0026] (3) Radiation exposure: the first time: 2Gy, the second time: 2Gy, the third time: 4Gy, the fourth time: 4Gy, the fifth time: 6Gy, the sixth time: 6Gy, the seventh time: 8Gy, the The eighth time: 8Gy, the ninth time: 10Gy, the tenth time: 10Gy; the eleventh time: 10Gy; the total radiotherapy dose is 70Gy; the cells formed after radiotherapy are named as radiotherapy resistant ...

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Abstract

The invention discloses a method for screening nasopharyngeal carcinoma radiotherapy resistant cells by using a radioactive irradiation mode. The method adopts a screening mode of radioactive gradient increasing. By adopting the method, cells can adapt to high-dose radioactive irradiation more rapidly within a relatively short time, the time of growth recovery after single high-dose radioactive irradiation can be shortened, the possibility of generating radiotherapy resistance can be increased, and meanwhile, the investment of manpower, material resources and time can be reduced. The constructed nasopharyngeal carcinoma radiotherapy resistant cells are used for providing a good cell model for the study on nasopharyngeal carcinoma radiotherapy resistance. The method disclosed by the invention is suitable for nasopharyngeal carcinoma cell strains cultured in vitro, and also can be used for providing references for the establishment of other tumor radiotherapy resistant cells.

Description

technical field [0001] The invention relates to a method for screening radiotherapy-resistant cells of nasopharyngeal carcinoma, in particular to a method for screening radiotherapy-resistant cells of nasopharyngeal carcinoma by radiation irradiation mode. Background technique [0002] At present, most of the construction methods of radiotherapy-resistant cells in nasopharyngeal carcinoma at home and abroad adopt the sublethal dose mode (10-12 Gy) to the tumor parental cells (such as "Identification of biomarkers for predicting nasopharyngeal carcinoma response to radiotherapy by proteomics" article;) or small-dose fractionated irradiation mode (1.8-2 Gy) (such as "The generation and characterization of a radiation" published in Volume 156 of Issue 6 of Radiat Res in 2001 -resistant model system to study radioresistance in human breast cancer cells "), but the sub-lethal dose screening mode has disadvantages such as difficult cell survival, easy denaturation and death of cel...

Claims

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Application Information

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IPC IPC(8): C12N13/00C12N5/09
Inventor 邱元正刘勇李果粟忠武任舒灵田勇泉
Owner XIANGYA HOSPITAL CENT SOUTH UNIV
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