The Research Of Inhibitory Effect Of EGFR Tyrosine Kinase Inhibitor On Lung Cancer Cells

Lung cancer has become the cancer with the highest morbidity and mortality.Studies have shown that overexpression of EGFR causes abnormal activation of downstream signals and are closely related to biological processes such as differentiation,proliferation,migration,and apoptosis of tumor cells.Non-small cell lung cancer has the phenomenon of overexpression of EGFR.As EGFR plays an important role in the formation of cancer,EGFR has been becoming an important target for anti-cancer therapy.Gefitinib and Erlotinib have been approved for marketing,but cancer cells have shown resistance to Gefitinib and Erlotinib,especially resistance caused by the T790M mutation.AZD9291 is currently available as an effective drug against T790M resistant mutations.However,at present,there are not many studies on the third generation EGFR inhibitors in China,and imported drugs are expensive,which results in the limited clinical application of drugs,and thus indirectly increase the mortality rate of lung cancer.In this study,lung cancer cell lines were used and target a series of synthetic compounds,compounds with strong T790M-mutant EGFR activity were screened out and studied in cell functions and mechanisms.1.The antitumor activity of quinazoline compounds 7a-7u was studied at the cell level,and the quinazoline compound 7d with high activity,high selectivity,and low toxicity were screened out.EGFR^L858R/T790M H1975 human lung adenocarcinoma cells,EGFR-wild-type H1563 human lung cancer cells and normal 16HBE human bronchial epithelial cells were selected.MTT assay was used to determine inhibitory activity of self-synthesis quinazoline compounds 7a-7u to lung cancer cells and toxicity of self-synthesis quinazoline compounds 7a-7u to normal cells 16HBE cells,using Erlotinib as a positive control drug,screening out highly active,highly selective,low toxicity quinazoline compound 7d as further research subjects.2.To study the inhibitory effect of the quinazoline compound 7d on proliferation,migration and invasion of lung cancer cellsH1975 cells and H1563 cells were used to study the inhibitory effect of quinazoline compound 7d on the proliferation,migration and invasion of lung cancer cells.The results showed that the quinazoline 7d could effectively inhibit the proliferation of H1975 cells using Erlotinib as a positive control drug.However,it had little effect on H1563 cells;it had a significant inhibitory effect on the migration and invasion of H1975 cells,but had little effect on the migration and invasion of H1563 cells.Mechanism studies have shown that the quinazoline compound 7d inhibits the invasion and migration of lung cancer cells by inhibiting the expression of the MMP2 and MMP9 proteins involved in invasion and migration.3.To investigate the mechanism of inhibition of H1975 cells by the quinazoline compound 7dH1975 cells were selected and flow cytometry was used to analyze the cell cycle distribution of H1975 cells before and after treatment with the quinazoline compound7d.It was found that the quinazoline compound 7d can block H1975 cells in G0/G1phase,and it was proved at the protein level.The quinazoline compound 7d down-regulated the expression of CyclinD1 and CyclinE1 proteins,which caused cell cycle arrest in G0/G1 phase.In summary,the quinazoline compound 7d has a good effect on EGFR T790M mutant lung cancer cells,not only can inhibit cell proliferation,but also can inhibit the invasion and migration of lung cancer cells by inhibiting the expression of the proteins MMP2 and MMP9 involved in invasion and migration.The quinazoline compound 7d can induce G0/G1 phase arrest by down-regulating the expression of CyclinD1 and CyclinE1 protein to induce G0/G1 phase arrest in H1975 cells.The quinazoline compound 7d has significant antitumor activity and good selectivity,and has good inhibitory activity against the T790M EGFR mutant non-small cell lung cancer.