Method for rapidly predicting and improving sensitivity of non-small-cell lung carcinoma cells to epidermis cell growth factor receptor inhibitor

A non-small cell lung cancer and growth factor receptor technology, applied in the biological field, can solve the problems of enhancing the sensitivity of EGFR inhibitors, unclear and lack of EGFR inhibitor resistance mechanisms, etc.

Active Publication Date: 2017-08-25
HANGZHOU BIOLYNX TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the resistance mechanism of more than 80% of NSCLC patients to EGFR inhibitors is still uncle

Method used

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  • Method for rapidly predicting and improving sensitivity of non-small-cell lung carcinoma cells to epidermis cell growth factor receptor inhibitor
  • Method for rapidly predicting and improving sensitivity of non-small-cell lung carcinoma cells to epidermis cell growth factor receptor inhibitor
  • Method for rapidly predicting and improving sensitivity of non-small-cell lung carcinoma cells to epidermis cell growth factor receptor inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Under cell culture conditions in vitro, the semi-lethal concentration (EC50) of 9 NSCLC cell lines treated with Erlotinib was determined by "concentration-effect" method.

[0031] experimental method:

[0032] 1.9 strains of NSCLC cells H322, H358, H441, Calu3, A549, H1703, SW1573, H460 and H23 were cultured with RPMI1640 medium (while adding 10% fetal bovine serum, 10 unit / ml penicillin and 10 unit / ml streptomycin). Culture conditions are 37°C, 5% CO 2 . Erlotinib was dissolved in DMSO at a stock solution concentration of 10 mM.

[0033] 2. Plant 10 cell lines at a density of 3000 cells / well on a 96-well cell culture plate. Twenty-four hours after cell seeding, the cells were treated with Erlotinib at concentrations of 10 μM, 5 μM, 2.5 μM, 1.25 μM, 0.625 μM, 0.3125 μM, 0.156 μM, 0.078 μM and 0 μM (DMSO). After 72 hours of treatment, CellTiter Glo reagent was used to measure cell activity, and Prism software was used to calculate the "concentration-effect" to obtain...

Embodiment 2

[0035] The data of eIF2α phosphorylation, eIF2α overall level, ATF4 expression level and internal reference GAPDH in 9 NSCLC cell lines were obtained by Western blotting. Then, the expression of the above proteins was correlated with the EC50 concentration of the respective cell lines to Erlotinib, and it was found that the levels of phosphorylated eIF2α and ATF4 were significantly correlated with the EC50 of cells to Erlotinib: the higher the phosphorylation level of eIF2α, the higher the expression level of ATF4 The higher the , the lower the sensitivity of lung cancer cells to Erlotinib (the higher the EC50).

[0036] experimental method:

[0037] 1.9 strains of NSCLC cells H322, H358, H441, Calu3, A549, H1703, SW1573, H460 and H23 were cultured with RPMI1640 medium (while adding 10% fetal bovine serum, 10 unit / ml penicillin and 10 unit / ml streptomycin). Culture conditions are 37°C, 5% CO 2 .

[0038] 2. Use standard RIPA buffer (20mM Tris-HCl (pH 7.5), 150mM NaCl, 1mM N...

Embodiment 3

[0050] Four cell lines H322, H441, A549, and H460 were planted subcutaneously in nude mice to form tumors, and treated with Erlotinib two weeks after implantation, and the data (TGI value) of the degree to which the four cell lines were inhibited by Erlotinib in the process of tumor formation were obtained. . Then the correlation analysis was carried out between the phosphorylation level of eIF2a and the expression level of ATF4 in the four cell lines and the inhibition degree of the four cell lines by Erlotinib in nude mice. The conclusion is that the phosphorylation level of eIF2a and the expression level of ATF4 are significantly correlated with the response of cells to Erlotinib in nude mice.

[0051] experimental method:

[0052] The method in Example 1 was adopted to culture, maintain and expand H322, H441, A549, and H460 cells. Collection 10 6 The cells were injected subcutaneously into the right groin of 8-week-old nude mice, and 10 nude mice were injected with each...

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Abstract

The invention provides a method for rapidly predicting and improving sensitivity of non-small-cell lung carcinoma cells to epidermis cell growth factor receptor inhibitor. The activation level of a PERK passage in UPR cells is detected, namely the phosphorylation degree of an eIF2a protein and the expression level of an ATF4 protein are detected, so that the sensitivity of an NSCLC to an EGFR inhibitor is predicted, and the activation level of the PERK passage in the UPR cells is suppressed, so that the sensitivity of the lung carcinoma cells to the EGFR inhibitor is improved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for rapidly predicting and improving the sensitivity of non-small cell lung cancer cells to epidermal growth factor receptor inhibitors. Background technique [0002] Lung cancer is a malignant tumor with the highest morbidity and mortality in the world. Although there are chemotherapy, radiotherapy, targeted therapy and immunotherapy at this stage, its 5-year survival rate is still less than 20% (Allemani C, Weir HK, Carreira H, et al. Global surveillance of cnacer survival 1995-2009: Lancel, 2015, 385(9972), 977-1010). Non-small cell lung cancer (NSCLC) is the most common type, accounting for about 80% (Lin Y, Wang X, Jin H. EGFR-TKI resistance on NSCLC patients, mechanisms and strategies, Am J Cancer Res, 2014, 4(5), 411-435). Epidermal growth factor receptor (EGFR) inhibitors are currently one of the most important drugs for targeted therapy of non-small cell lung canc...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/574
CPCG01N33/57423G01N33/6803G01N2333/4704G01N2800/12
Inventor 冯宇雄郭箭
Owner HANGZHOU BIOLYNX TECH CO LTD
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