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Anti-crizotinib human non-small cell lung cancer cell strain H3122-CR23 and application thereof

A technology of H3122-CR23 and non-small cell lung cancer, which is applied in the field of tumor biology to achieve high scientific research and production application value

Active Publication Date: 2017-10-20
THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In order to explore the mechanism of acquired resistance to crizotinib, we established a crizotinib-resistant non-small cell lung cancer cell line H3122-CR23, which has a F1174C mutation in the ALK kinase region, which has not been seen at home and abroad There are reports of such crizotinib-resistant non-small cell lung cancer cell lines

Method used

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  • Anti-crizotinib human non-small cell lung cancer cell strain H3122-CR23 and application thereof
  • Anti-crizotinib human non-small cell lung cancer cell strain H3122-CR23 and application thereof
  • Anti-crizotinib human non-small cell lung cancer cell strain H3122-CR23 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Preparation of crizotinib-resistant human non-small cell lung cancer cell line H3122-CR23

[0021] The steps are: (1) Human ALK-positive non-small cell lung cancer cell line H3122 (hereinafter referred to as human non-small cell lung cancer cell line H3122) was purchased from the American Type Culture Collection (ATCC). Place the cell H3122 strain in RPMI1640 culture medium containing 25 mmol / L 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) and 10% fetal bovine serum (FBS) (hereinafter referred to as H3122 medium) at 37°C, 5% CO 2 , cultured in an incubator with saturated humidity, digested and passaged with 0.25% trypsin and ethylenediaminetetraacetic acid (0.25% Trypsin-EDTA);

[0022] (2) Take the H3122 cells in the logarithmic growth phase or when the H3122 cells are cultured until the cell density reaches 70%, change the fresh H3122 medium, add crizotinib to form a culture solution containing crizotinib, which contains crizotinib The concent...

Embodiment 2

[0024] Example 2 Cryopreservation of crizotinib-resistant human non-small cell lung cancer cell line H3122-CR23 of the present invention

[0025] The drug-resistant cells obtained in Example 1 were placed in the cryopreservation solution of the cryopreservation tube, and then the cryopreservation tube was placed in liquid nitrogen for preservation. The cryopreservation medium consists of 90% H3122 medium and 10% dimethyl sulfoxide (DMSO). The cryopreservation process takes the steps of placing the cryopreservation tube in a nalgene programmed cooling box, overnight at -80°C, and finally putting it into liquid nitrogen.

Embodiment 3

[0026] Example 3 Recovery of crizotinib-resistant human non-small cell lung cancer cell line H3122-CR23

[0027]Take out the cryopreservation tube containing the cells from the liquid nitrogen, immediately put it in a 37-40°C water bath and shake it gently, so that the frozen storage will thaw within 1 minute, wipe the outer wall of the cryopreservation tube with an alcohol cotton ball and put it into the ultra-clean tower. Place the thawed cell suspension in a 15ml sterile centrifuge tube, add 5ml of phosphate buffered saline (PBS buffer), centrifuge at 1000 rpm for 5 minutes, discard the supernatant, add 5ml of crizotinib containing 1000nmol / L H3122 culture medium, slightly pipetting and mixing to form a mixed solution, suck the mixed solution and put it into a 25cm 2 In the cell culture bottle, unscrew the bottle cap and put it in a carbon dioxide incubator at 37°C, 5% CO 2 1. Cultivate under the condition of saturated humidity, replace the medium once every 3-4 days, dig...

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Abstract

The invention discloses a novel anti-crizotinib human non-small cell lung cancer cell strain which is named as H3122-CR23 and has an accession number of CCTCC No. C201780. The cell strain has F1174C mutation in an ALK kinase area. The cell strain H3122-CR23 is applicable to research on the morphological and biological characteristics of human anti-crizotinib non-small cell lung cancer cells, research on the drug resistance mechanisms of tumors, development of drugs for drug resistance reversal of tumors, analysis of the susceptibility of antitumor drugs, screening and assessment of antitumor drugs, research on more efficient tumor treatment methods, etc., has high application value in scientific research and production, and is expected to produce good scientific research, economic and social benefits.

Description

technical field [0001] The invention relates to the field of tumor biology, in particular to a crizotinib-resistant human non-small cell lung cancer cell line H3122-CR23, and also relates to the application of the non-small cell lung cancer cell line H3122-CR23. Background technique [0002] Lung cancer is a common malignant tumor that seriously endangers human life and health. Lung cancer is divided into small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC), of which 80% to 85% are non-small cell lung cancer. Traditional treatment for NSCLC includes surgery, chemotherapy, and radiation. With the in-depth study of tumor pathogenesis and its biological behavior, people are now focusing on molecular targeted therapy with high specificity and mild adverse reactions. [0003] In recent years, the field of molecular targeted therapy for lung cancer has focused on targets such as EGFR, K-ras and VEGF, among which EGRF targeted therapy has achieved gratifying resul...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09A61K35/13A61P35/00C12Q1/02
CPCA61K35/13C07K14/71C12N5/0688C12N5/0693G01N33/5044
Inventor 胡野荣周杨钊周新民
Owner THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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