Brigatinib Induced The PI3K/AKt/mTOR Signaling Pathway In The Progression Of NSCLC

Background: Lung cancer is one of the main cause of death from cancer worldwide.The prognosis of advanced lung cancer is very bad.As the limitation of traditional chemotherapy and radiotherapy,the new targeted gene therapy has become a new research direction of NSCLC.In the last few years,ALK inhibitors,such as crizotinib,ceritinib and alectinib have become the treatment of EML4-ALK fusion gene positive NSCLC patients.The new generation of ALK inhibitors,Brigatinib,has shown a striking prospect in the EML4-ALK positive NSCLC patients.Objective:To investigate the role of PI3K/AKT/mTOR signaling pathway and cell growth in EML4-ALK fusion gene positive NSCLC induced by Brigatinib.Methods : H2228 cells were processed with different concentrations of Brigatinib according to distinct requirements.Brigatinib-induced H2228 cell inhibition rates were tested by MTT assay.Brigatinib-induced apoptosis inhibition of H2228 at different doses of Brigatinib was measured using flow cytometry test.After the treatment of Brigatinib,the key genes mRNA and protein levels of p-ALK,p-PI3K,p-AKt,and p-mTOR in H2228 were detected by qRT-PCR technique and Western blot Results: By MTT test,IC50 values for H2228 were 115.9nM,the proliferation of H2228 were inhibited by Brigatinib,and had dose-dependent,with a statistically significant difference(p<0.05).Brigatinib can siganificantly reduced H2228 cells apoptosis by flow cytometry detection,and have dose-dependent,and the cell line was blocked in G1 phase,compared with control group,there is a statistical difference(p<0.05).Western blot and qRT-PCR showed that Brigatinib significantly inhibits the expression of p-ALK,p-PI3K,p-AKt,and p-mTOR in H2228 treated by different doses of Brigatinib,compared with control group,there is a statistical difference(p<0.05).Conclusions: In conclusion,Brigatinib promotes H2228 cells proliferation and inhibits cell apoptosis,in a dose-dependent.Bragatinib adjust H2228 cells proliferation and apoptosis by down-regulating p-ALK,p-PI3K,p-AKT and p-mTOR.It indicates that PI3K/AKT/mTOR signaling pathway has potential clinicalsignificance in the progression of Brigatinib-induced EML4-ALK+ NSCLC.