Dual mTORC1/2 inhibitor AZD2014 inhibits proliferation of HCCLM3 cells through induction of autophagy

Objective To investigate the effect of mTORC1/2 inhibitor AZD2014 induced autophagy on cell proliferation in HCCLM3 cell lines in vitro. Methods HCCLM3 cells were cultured in the presence or absence of AZD2014 (100 nmol/L) or rapamycin (100 nmol/L) or 3-methyladenine (10 nmol/L). To detect autophagy vacuoles, HCCLM3 cells were divided into the control group, rapamycin group, AZD2014 group, control+3-MA group, rapamycin+3-MA group, and AZD2014+3-MA group, and auto-fluorescent dye monodansylcadaverine (MDC) was used to monitor autophagy vacuoles. To semi-quantify the expression of autophagy-related proteins, HCCLM3 cells were divided into the control group, rapamycin group, and AZD2014 group, and Western blotting analysis was carried out to detect the expression of autophagy-related proteins, LC3B-II and Beclin-1. To evaluate the effect of autophagy induced by AZD2014 on cell proliferation, HCCLM3 cells were divided into the control group, 3-MA group, AZD2014 group, and AZD2014+3-MA group, Cell Counting Kit-8 was used. Results AZD2014-treated HCC cells showed an increase in the number of MDC-labeled vacuoles as well as in their size. AZD2014 treatment increased the levels of LC3B-Ⅱ and Beclin-1 (P