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Osimertinib drug-resistant cell line NCI-H1975/AR and application thereof

A technology of NCI-H1975 and drug-resistant cells, which is applied in the field of biomedicine to achieve an important role and a good application prospect.

Active Publication Date: 2020-06-02
THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, many drug-resistant cell lines of tumors have been established at home and abroad, but there is no first-line drug-resistant cell line of non-small cell lung cancer to AZD9291

Method used

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  • Osimertinib drug-resistant cell line NCI-H1975/AR and application thereof
  • Osimertinib drug-resistant cell line NCI-H1975/AR and application thereof
  • Osimertinib drug-resistant cell line NCI-H1975/AR and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Induction and establishment of embodiment 1 drug-resistant cell line

[0022] Induction construction of drug-resistant human lung adenocarcinoma cell line NCI-H1975 / AR:

[0023] (1) Culture non-small cell lung cancer cells H1975 in a medium containing 10% inactivated fetal bovine serum, 1% double antibody (100 U / ml penicillin, 100 μg / ml streptomycin), and place at 37°C, 5% CO 2 1. Cultivate in an electric heating constant temperature incubator under the condition of 96% humidity;

[0024] (2) The non-small cell lung cancer cells in the logarithmic growth phase were divided into 5×10 5 / mL inoculated in a culture flask, and when cultured to 75% adherence rate, discard the supernatant, add 100nM AZD9291 drug culture solution to continue to act on the cells, passage when the cell growth accounted for 80-90% of the culture dish, and aspirate For the old culture medium, add 2ml PBS to wash once, add an appropriate amount of trypsin-EDTA digestion solution, and then wash wi...

Embodiment 2

[0026] Morphological observation of embodiment 2 drug-resistant cell lines

[0027] 1. Material of test cell line:

[0028] The non-small cell lung cancer drug-resistant cell line constructed in Example 1, that is, the human lung adenocarcinoma drug-resistant cell line NCI-H1975 / AR, and the parental non-small cell lung cancer cell line H1975 / pt.

[0029] 2. Test method:

[0030] The two kinds of cells were seeded in 6-well plates, cultured to the logarithmic growth phase, and the cell morphology was observed under an inverted phase-contrast microscope.

[0031] 3. The test results are as follows: figure 1 shown.

[0032] Under the microscope, both H1975 / pt and H1975 / AR cells showed epithelioid single-layer arrangement and adherent growth. H1975 / pt cells had clear borders, mostly long fusiform and even polygonal, and the growth was loose with gaps; while H1975 / AR after drug resistance The cells became larger in size, with ill-defined borders, gathered and grown in clusters,...

Embodiment 3

[0034] The detection of drug sensitivity and drug resistance of embodiment 3 drug-resistant cell lines

[0035] 1. Material of test cell line:

[0036] The non-small cell lung cancer drug-resistant cell line constructed in Example 1, that is, the human lung adenocarcinoma drug-resistant cell line NCI-H1975 / AR, and the parental non-small cell lung cancer cell line H1975 / pt.

[0037] 2. Test method:

[0038]The logarithmic non-small cell lung cancer drug-resistant cell line H1975 / AR and the parental non-small cell lung cancer cell line H1975 / pt were respectively prepared into cell suspensions with a concentration of 30,000 cells / ml, and seeded in 96-well cell culture plates, each well Add 200 μl of cell suspension (6000 cells); place the 96-well cell culture plate at 37°C, 5% CO 2 , in an incubator with a humidity of 96%, after culturing the cells for 24 hours to adhere to the wall, carefully suck out the old medium, and add 200 μl of prepared drug-containing medium solutions ...

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Abstract

The invention discloses an osimertinib drug-resistant cell line NCI-H1975 / AR and an application thereof. The drug-resistant cell line is a non-small-cell lung cancer-resistant AZD9291 cell line, namely human lung adenocarcinoma cell NCI-H1975 / AR, the drug-resistant cell line is preserved on June 14, 2019 at China Center for Type Culture Collection, the preservation number is CCTCC NO:C201982. Thedrug resistance index of the drug-resistant line is 10 times or more, and the drug-resistant line can be stably grown and passaged in a culture system with an action concentration of 100 nmol AZD9291,the drug resistance index (RI) to the AZD9291 is 17.7 times, and the drug-resistant cell line provides a drug-resistant cell model for the drug-resistance mechanism of non-small cell lung cancer to atargeted drug, finding of effective treatment methods to overcome drug resistance in the non-small cell lung cancer, and guiding of clinical medication, and has important effects and good applicationprospects.

Description

technical field [0001] The invention belongs to the technical field of biomedicine. More specifically, it relates to an osimertinib-resistant cell line NCI-H1975 / AR and its application. Background technique [0002] Lung cancer ranks first in cancer mortality worldwide, and non-small-cell lung cancer (NSCLC) accounts for 80% of lung cancer. The 5-year survival rate of non-small cell lung cancer is less than 15%. The main reason is that about 70% of patients with non-small cell lung cancer are already at an advanced stage when they are first diagnosed with distant metastasis. Lung cancer is one of the first types of cancer to use targeted therapy in the clinic. In the past decade, epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) has been an important drug for the treatment of advanced non-small cell lung cancer (NSCLC), and its target is Epidermal Growth Factor Receptor (EGFR). This type of drug has the characteristics of definite curative effect, mil...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/02
CPCC12N5/0693G01N33/5011G01N2500/10
Inventor 李谨
Owner THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT)
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