Effect Of GNAI2 On Biological Function Of Prostate Cancer Cell PC-3

Objectives To investigate the effect of silenceing GNAI2 on proliferation,migration,cell cycle and mitochondrial membrane potential of prostate cancer cells PC-3.Methods Recombinant plasmids capable of silencing GNAI2 gene and its control recombinant plasmid were constructed.Lentivirus in which two packaging were used contain GNAI2 si RNA and its control were packaged by PEI.This Two lentiviruses were transfected into PC-3 cells for constructing cell model which silencing expressed GNAI2 and its control group which called.Cells were divided into three groups: 1 PC-3 control(NC),2 Knockdown control group(SHCON),3 Knockdown group(SHGNAI2).Cell proliferation ability was detected by MTT assay.Cell migration ability was detected by cell scratch test.Cells of each groups were stained by PI(Propidium Iodide,PI)for detecting cell cycle on Flow cytometer.Cells of each groups were stained by TMRE(Tetramethylrhodamine ethyl ester,TMRE)for detecting mitochondrial membrane potential on Flow cytometer.Results 1 After identification,the recombinant plasmids of knock-down group and its control group were successfully constructed and meanwhile the corresponding lentivirus were also packaged successfully.2 Following the cell models of PC-3 silencing GNAI2 and its control group were constructed successfully.3 The MTT assay showed that silencing GNAI2 Promoted Proliferation of PC-3 Cells.There was no significant differences among each groups at the 24 th hour(P>0.05).At the 48 th hour,the proliferation of cells in each group was appropriately increased.The difference between SHGNAI2 group and the two control groups was significant(P<0.05).There was no significant difference between NC group and SHCON group(P=0.214).At the 72 th hour,the proliferation of the three groups of cells was more,and the difference between the SHGNAI2 group and the two control groups was significant(P<0.001).There was no significant difference between the NC group and the SHCON group(P=0.193).4 It inhibited the migration of PC-3 cells that GNAI2 gene silencing.The repair rate of scratches in each group at 24 th hour were as follows: LV-SHGNAI2(16.99±1.65),SHCON(57.67±3.17),NC(54.43±2.12).The cell migration ability of LV-SHGNAI2 group was significantly lower than that of the two control groups,and the repair rate was smaller than that of the two control groups(P<0.001).There was no significant difference between NC group and SNCON group(P=0.074).5 Silencing of GNAI2 made PC-3 cell proliferate.The distribution of cell cycle of SHCON group and SHGNAI2 group are as follows: SHCON: G0/G1 phase(55.90 ± 1.72),S phase(32.48 ± 3.66),G2/M phase(8 ± 0.2).SHGNAI2: G0/G1 phase(57.62±0.67),S phase(41.27±1.73),G2/M phase(1.8±1.68).Compared with SHCON group,the proportion of cells in S phase of SHGNAI2 group increased(P<0.05),the proportion of cells in G2/M phase decreased(P<0.05),and the proportion of cells in G0/G1 phase did not change(P=0.224).6 The silence of GNAI2 increased the Mitochondrial Membrane Potential in PC-3 Cells.Mitochondrial membrane potential of each group is as follows: SHCON(71.39±1.23),SHGNAI2(91.08±2.8).Conclusions Silencing GNAI2 promoted the proliferation of PC-3 cells,inhibited the migration of PC-3 cells.increased PC-3 S phase cells,and increased the mitochondrial membrane potential of PC-3 cells which suggesting that GNAI2 plays an critical role in the progression of PCa.