Study Of Effects And Mechanism Of MTOR Inhibitors Everolimus On Human Prostate Cancer PC-3Cell Lines In Vitro

Objective: To research of changes of mTOR inhibitors Everolimus(RAD001) on proliferation of human prostate cancer PC-3cell lines in vitro,and search for its possible mechanism.Methods:Human prostate cancer PC-3cell lines were cultivated in vitro by Cellculture technology and then interfered with by1nmol/l Everolimus (RAD001).Changes of Everolimus (RAD001) on the proliferation of PC-3cell lines wasdetected after Everolimus interfere with PC-3cell lines for24hours,36hours,48hours,72hours by MTT assay. The influence of Everolimus (RAD001) onthe cell cycle change was detected after Everolimus interfere with PC-3celllines for24hours,36hours,48hours,72hours by Flow cytometry analysis(FCM). The changes of Everolimus (RAD001) on expression of p-4E-BP1andp-P70-S6K was detected after Everolimus interfere with PC-3cell lines for24hours,36hours,48hours,72hours by Western blot analysis. Experimentaldates were analyzed by SPSS13.0, and P <0.05is defined as a Statisticallysignificant difference.Results:1MTT assays results suggested that Everolimus (RAD001) can inhibit theproliferation of PC-3cell lines, and with the extension of drug interventiontime, the inhibition rate rise gradually (P <0.01).2The results of Flow cytometry analysis (FCM) revealed that after humanprostate cancer PC-3cell lines were treated with by Everolimus (RAD001),with the extension of drug intervention time, the cell proportion of G0/G1phase rises gradually, the cell proportion of S, G2/M phase declinegradually，and make the most of the PC-3cells are interrupted in G0/G1phase of the cell cycle (P <0.01). 3Western blot analysis showed that after human prostate cancer PC-3celllines were interfere with by Everolimus (RAD001), with the extension of drugintervention time, the expression of p-4E-BP1and p-P70-S6K were graduallysuppressed (P<0.01).Conclusions:mTOR inhibitors Everolimus (RAD001) can inhibit the proliferation ofPC3cell lines, its mechanism may be related to the mTOR suppressed ofPI3K/Akt/mTOR signaling pathways, and then which inhibits two importantproteins (i.e., translation inhibitory factor eIF-4E binding protein1(4E-BP1)and ribosomal protein P70-S6K) of its downstream phosphorylation.