Mechanisms Of 16-Acetoxy-7-O-Acetylhorminone Inhibition Of Proliferation Of EGFR Wild-Type Lung Cancer Cells

One of the important therapeutic targets for tumors is the epidermal growth factor receptor(EGFR),the activation of the EGFR signaling pathway promotes tumor growth and progression,including angiogenesis,invasion,metastasis,and inhibition of apoptosis.85% of lung cancer patients have non-small cell lung cancer(NSCLC),and EGFR is frequently overexpressed in NSCLC.Currently,several therapeutic agents have been developed for the treatment of EGFR mutant NSCLC lung cancer,such as tyrosine kinase inhibitors,monoclonal antibodies,as broad-spectrum anti-EGFR inhibitors,all of which have good therapeutic effects on lung cancer cell lines with abnormal EGFR expression,and the introduction of these drugs has prolonged the progression-free survival of lung cancer patients and increased their five-year survival rate.However,there are few types of drugs targeting EGFR wild-type NSCLC lung cancer,as well as poor therapeutic efficacy.16-Acetoxy-7-O-acetylhorminone(16-A)is an active component of chytrid,but its physiological activity is not yet known.The purpose of this study was to evaluate the anticancer activity of 16-A in EGFR WT-type NSCLC and to investigate its mechanism of action.Experimental studies were performed by MTT assay,cell morphological observation,scratch assay,cell colony formation assay,flow cytometry and Western Blot assay methods.The results of the experimental study were as follows: 16-A dose-dependently inhibited the proliferation of three EGFR wild-type NSCLC cell lines;16-A significantly changed the morphology of EGFR wild-type cell lines NCI-H292,NCI-H1299 and NCIH2073 cells;it also inhibited cell migration.The migration ability of NCI-H1299 cells was lost when treated with 16-A 2 μmol/L,and the migration ability of NCI-H2073 cells was lost when treated with 16-A 8 μmol/L.16-A inhibited cell clone formation,and the colony formation ability of NCI-H292 cells was lost by 70% when treated with 16-A 4 μmol/L,and the colony formation ability of NCI-H1299 cells was lost by 70% when treated with16-A 4 μmol/L.NCI-H1299 cells lost 50% of their colony-forming ability when treated with 16-A at 4 μmol/L,and NCI-H2073 cells lost 80% of their colony-forming ability when treated with 16-A at 10 μmol/L.Flow assay of cell cycle showed that 16-A could block the cell cycle of NCI-H292 cells and block the cycle in S/G2 phase;flow assay of apoptosis showed that 16-A could promote apoptosis in three cell lines and increase the rate of apoptosis;Western Blot assay of the effect of 16-A on EGFR signaling pathway protein expression showed that 16-A could inhibit the expression of EGFR signaling pathway protein in NCI-H292 The results showed that 16-A could inhibit the phosphorylation of EGFR and ERK(extracellular signal-regulated kinase)in NCI-H292 cells and block the signal transduction;Western Blot examined the expression of cell cycle proteins,and 16-A could promote the expression of cell cycle-dependent kinase inhibitor protein,and then inhibit the expression of cell cycle protein-dependent kinase,thus inhibiting the expression of cell cycle proteins and Western Blot assay of apoptotic protein expression,16-A can inhibit the expression of anti-apoptotic protein Bcl-2(B-cell lymphoma-2)and promote the expression of pro-apoptotic protein Bax(Bcl-2-related X protein),which in turn promotes the expression of Caspase family proteins,initiates the protein hydrolysis cascade reaction,causes apoptosis and inhibits cell proliferation.In summary,16-A inhibits the proliferation of EGFR wild-type NSCLC lung cancer cells,and 16-A induces apoptosis by regulating the expression of apoptosis proteins and cyclins to inhibit the proliferation of cancer cells.16-A’s discovery initially provides new therapeutic drugs and therapeutic directions for the treatment of EGFR wild-type NSCLC lung cancer patients,and also provides a new basis for the research and development of natural compounds.development.