The Effects Of Knock-down STOML2 Gene On Biological Behavior Of Prostate Cancer PC3 Cells

Objectives Knocked down the Stomatin-like protein 2(STOML2)gene of PC3 cells by lentiviral technology,we observed the effects on the proliferation,migration,invasion,the level of Erk and Akt pathway expression,mitochondrial membrane potential and reactive oxygen species of prostate cancer PC3 cells.Methods Human prostate cancer cells PC3 and HEK293 T were selected,and HEK293 T cells were transfected by PEI and provided knocked down the STOML2 gene and random sequence lentivirus,which can infect prostate cancer cell PC3.Immunofluorescence microscopy and Western Blot(WB)were used to detect infection efficiency and STOML2 protein expression level.The proliferation,migration and invasion of PC3 cells were detected by MTT assay,cell scratch healing assay and Transwell assay.WB and flow cytometry were used to detect Erk,Akt protein expression level,mitochondrial membrane potential,and reactive oxygen species in PC3 cells after knocked down STOML2 gene.Results Two lentiviral plasmids,sh DNA-STOML2 and random sequence sh DNA,were successfully constructed.The infection efficiency of SHCON group and SHSTOML2 group was closed to 100%.After PC3 cells infected,the relative expression levels of STOML2/?-actin in Wt group,SHCON group and SHSTOML2 group were(0.262 ±0.021),(0.247 ±0.013)and(0.069±0.013),and the difference between SHSTOML2 and SHCON group was statistically significant(P<0.01).It indicated that the model was successfully constructed.Compared with SHCON group,the SHSTOML2 group increased cell viability at 24 h,48h and 72h(P<0.01),decreased cell migration and invasion ability at24 h and 48h(P<0.01).Compared with SHCON group,knocked down STOML2 gene reduced p-erk and Akt protein expression,but increased p-akt protein expression(P<0.05).Compared with SHCON group,the SHSTOML2 group increased mitochondrial membrane potential and reactive oxygen species at 48h(P<0.01).Conclusions Knocked down of the STOML2 gene can accelerate PC3 cell proliferation and decrease migration and invasion ability,which may be related to the increased mitochondrial membrane potential,reactive oxygen species and activation of Akt and reduction of Erk pathway in PC3 cells.Figure 11;Table 3;Reference 139...