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A radiotherapy-resistant lung cancer cell line and its construction method and application

A lung cancer cell, radiotherapy-resistant technology, applied in the direction of tumor/cancer cells, animal cells, vertebrate cells, etc., can solve the problem of lack of radiotherapy-resistant lung cancer cell lines, etc., achieve good application prospects, strong radiotherapy tolerance, Radiation Tolerant and Stable Outcomes

Active Publication Date: 2020-03-27
HANGZHOU FIRST PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Aiming at the problem of lack of radiotherapy-resistant lung cancer cell lines in the prior art, the present invention provides a radiotherapy-resistant lung cancer cell line and its construction method and application

Method used

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  • A radiotherapy-resistant lung cancer cell line and its construction method and application
  • A radiotherapy-resistant lung cancer cell line and its construction method and application
  • A radiotherapy-resistant lung cancer cell line and its construction method and application

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Experimental program
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Effect test

Embodiment 1

[0024] 1. Cell culture: use RPMI-1640 medium (which contains 10% fetal bovine serum, 100U / ml penicillin, 100 μg / ml streptomycin and 2mM glutamine), and placed in 5% CO 2 , Cultured in a constant temperature incubator at 37°C.

[0025] 2. Cell irradiation: Cells in the exponential growth phase are irradiated by 6MV X-rays from the Swedish Elekta Precise linear accelerator, the source-to-skin distance is 100cm, the irradiation field is 30cm×30cm, and the dose rate is 500Gy / min. compensation block.

[0026] 3. Establishment of radiotherapy-resistant cell line PC9-RR: Human lung adenocarcinoma cell line PC9 (purchased from the Cell Resource Center, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences) cells (about 35×10 4 1) were inoculated in T25 culture flasks, and were irradiated by X-Ray 4Gy when the cells grew to 60% density; fresh culture medium was replaced immediately after irradiation, and the medium was changed every 2 days. Continue to culture unti...

Embodiment 2

[0028] Observation of cell morphology: Take monolayer cells in the exponential growth phase and observe their morphological differences under an inverted phase-contrast microscope.

[0029] refer to figure 1 As shown, under the phase-contrast microscope, the normal growth of PC9 and PC9-RR cells were similar in shape, and they both showed monolayer adherent growth. Among them, PC9 cells were relatively uniform in size, and PC9-RR elongated spindle cells increased.

Embodiment 3

[0031] Cell proliferation kinetics: Take 1×10 PC9 and PC9-RR cells each 4 Inoculated in 24-well plates (21 wells each), digested and counted the cells after inoculation and culture for 24h, 48h, 72h, 96h, 120h, 144h, and 168h respectively, counted the cells in 3 wells at each time node, and calculated the average value to draw the cell growth curve, and calculate the population doubling time PDT=t×lg2 / (1gN t -lgN 0 ), where N0 is the initial cell number, Nt is the final cell number, and t is N t -N 0 time.

[0032] refer to figure 2 As shown, the cell doubling time was calculated according to the growth curve, and the PC9 and PC9-RR cells were 34h and 37h respectively, and the PC9-RR cell doubling time was longer than that of the parental cell PC9.

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Abstract

The invention discloses a radiotherapy tolerance lung cancer cell line, a construction method thereof and an application of the cell line. The radiotherapy tolerance lung cancer cell line is named asa human lung adenocarcinoma radiation tolerance cell strain PC9-RR, and the collection number is CGMCC No. 14314. The radiotherapy tolerance lung cancer cell line comprises radiotherapy tolerance cells acquired by performing repeated X-ray irradiation and culture on a human lung adenocarcinoma cell strain PC9, is strong and stable in radiotherapy tolerance and has a good application prospect in terms of serving as a cell model for lung cancer radiotherapy resistance mechanism research and the like.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a radiotherapy-resistant lung cancer cell line and its construction method and application. Background technique [0002] Lung cancer is one of the malignant tumors that seriously endanger human health. The overall prognosis is poor, and the 5-year survival rate is about 17%. At present, radiation therapy is still an important part of the comprehensive treatment of lung cancer, and about 60%-70% of lung cancer patients need to receive radiation therapy at different stages of the disease. With the continuous improvement of imaging and radiotherapy technology, radiotherapy is widely used as a main radical method for inoperable stage I, II, III, and limited stage IV patients. For stage IV patients with multiple metastases, the targeted and immune drug therapy that has appeared in recent years has also brought more survival time and opportunities for patients with advanced lung cancer, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09C12N13/00C12Q1/02A01K67/027
CPCA01K67/0271A01K2207/12A01K2227/105A01K2267/03C12N5/0693C12N13/00C12N2503/02G01N33/5011G01N2500/10
Inventor 张仕蓉张静静赵妍妍马胜林吴琼许雅思黄海琇
Owner HANGZHOU FIRST PEOPLES HOSPITAL
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