Construction method of lung cancer radiation resistance cell strain
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A technology of radiation resistance and construction method, which is applied in the field of cell biology to achieve the effect of less chance of cell contamination, shorter time, and fewer times of irradiation
Inactive Publication Date: 2015-10-07
XUZHOU MEDICAL COLLEGE
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However, at present, the total dose of conventional dose-fractionated radiotherapy in clinical practice should not be less than 60G
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Embodiment 1
[0024] like figure 1 As shown, a method for constructing a radiation-resistant cell line of lung cancer includes the following steps:
[0025] ①The human lung cancer cell line A549 was placed in 37℃, 5%CO 2 Conventional culture in the incubator, when it covers 60-70% of the culture flask and is in the logarithmic growth phase, with 5 × 10 per well 5 Place the six-well plate in the center of the irradiation field, cover the bottom surface of the six-well plate with 1.5cm thick tissue equivalent filler, and the irradiation source is Varian23-EX medical linear accelerator, 6MV-X-ray, rack Rotate 180°, the rays are vertically incident from below at room temperature, the dose rate is 300cGy / min, the distance from the source to the skin is SSD=100cm, the size of the irradiation field is 30×30cm, and the first irradiation is 2Gy;
[0026] ②Replace the fresh medium after irradiation, when the cells cover more than 80% of the culture flask, trypsinize, and passage for more than two t...
Embodiment 2
[0037] A method for constructing a radiation-resistant cell line of lung cancer, comprising the following steps:
[0038] ①Put the human lung cancer cell line H1299 at 37℃, 5%CO 2 Conventional culture in the incubator, when it covers 60-70% of the culture flask and is in the logarithmic growth phase, with 5 × 10 per well 5 Place the six-well plate in the center of the irradiation field, cover the bottom surface of the six-well plate with 1.5cm thick tissue equivalent filler, and the irradiation source is Varian23-EX medical linear accelerator, 6MV-X-ray, rack Rotate 180°, the rays are vertically incident from below at room temperature, the dose rate is 300cGy / min, the distance from the source to the skin is SSD=100cm, the size of the irradiation field is 30×30cm, and the first irradiation is 2Gy;
[0039] ②Replace the fresh medium after irradiation, when the cells cover more than 80% of the culture flask, trypsinize, and passage for more than two times;
[0040]③ After passa...
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Abstract
The invention discloses a construction method of a lung cancer radiation resistance cell strain. The method includes the following steps of inoculating lung cancer cells in a six-hole plate when the lung cancer cells are conventionally bred to the logarithmic phase, placing the six-hole plate at the center of an irradiation field, covering the bottom face of the six-hole plate with tissue equivalent filler 1.5 cm thick, perpendicularly transmitting 6MV-X rays according to the dose rate of 300 cGy/min and the source skin distance of 100 cm, sequentially conducting irradiation according to the dose of 2 Gy, 4 Gy, 6 Gy, 8 Gy and 10 Gy each for two times till the total dose reaches 60 Gy, continuing to conduct breeding and passage to five or more generations till the cellular morphology and multiplication are stable, and conducting cryopreservation for standby application. The construction method is small in irradiation time number, short in construction time, small in cell contamination probability and superior to a small-dose equal-segmentation irradiation method; the colony formation experimental result shows that the radiation resistance of the lung cancer radiation resistance cell strain constructed according to the method is higher than that of lung cancer radiation resistance cell strains constructed according to the sublethal dose irradiation method.
Description
technical field [0001] The invention belongs to the field of cell biology, and in particular relates to a method for constructing a radiation-resistant cell line of lung cancer with stable radiation resistance. Background technique [0002] Lung cancer is one of the most common malignant tumors in my country. In 2014, the National Cancer Center released the "Annual Report on Cancer Registration in China", which reported that lung cancer ranks first in the incidence of malignant tumors in my country, with about 600,000 new cases and 49 deaths each year. Ten thousand. Radiation therapy is one of the most important means of multidisciplinary comprehensive treatment of lung cancer. Statistics show that more than 70% of patients receive radiation therapy during the diagnosis and treatment process. However, despite comprehensive treatment by various means, the overall efficacy of lung cancer is unsatisfactory, and the 5-year survival rate has been hovering around 15% for a long ti...
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