ABSTRACT
Objective To explore the effect and molecular mechanism of miR-34c in cell proliferation and invasion of human prostate cancer PC3 cells.Methods miR-34c was synthesized and transfected into prostate cancer PC3 cells.The ability of cell pro liferation was examined with methyl thiazolyl tetrazolium (MTT),and invasion with Transwell assay.The level of c-met mRNA was analyzed by real-time reversing transcription polymerase chain reaction (RT-PCR).Western blot was used to detect the expressions of cmet,cyclin D1,and EMT-associated markers.Results Over-expression of miR-34c reduced PC3 cell proliferation,and inhibited the abilities of invasion.Bioinformatics analysis showed that 3'-UTR of c-met had two miR-34c matched sites,and miR-34c inhibited the expressions of c-met mRNA and protein.Meantime,miR-34c obviously decreased the expressions of cyclin D1 and N-cadherin,but increased the expression of E-cadherin in PC3 cells.Conclusions miR-34c may suppress cell proliferation and invasion in prostate cancer PC3 cells,by which mechanism is related to reduction of the expressions of c-met,cyclin D1,N-cadherin,and upregulation of E-cadherin.
ABSTRACT
Objective To investigate the antitumor activities of adenovirus-mediated NDRG2 gene (Ad-NDRG2) and docetaxel on human prostate cancer DU145 cells.Methods The protein expressions of cyclin D1,cycliu E,and NDRG2 in the cells were determined by Western blot.MTT and flow cytometry were used to observe the effects of docetaxel (10-6 mol/L,10-7 mol/L,and 10-s mol/L) and Ad-NDRG2 on prostate cancer cell line DU145 in single or synergistic administration ways for 24 and 48 hours in vitro.Male BALB/C-nu mice with DU145 prostate cancer cell lines were treated by docetaxel and Ad-NDRG2 singly or synergistically in vivo.Results After infected by adenovirus,the protein expression of NDRG2 increased,but cyclin D1 and cyclin E decreased in DU145 cells.Ad-NDRG2 inhibited the cell growth (inhibition ratio =41.8％,t =4.18,P ＜0.01),promoted apoptosis (apoptosis ratio =32.4％,x2 =11.66,P ＜0.05),changed the ratio of G2/M phase from 50.2％ to 23.6％,and reversed partially the G2/M arrest,of DU145 cells induced by 10-7 mol/L docetaxel.In vivo experiment showed that docetaxel,Ad-NDRG2,and combination of docetaxel and Ad-NDRG2 inhibited tumor growth with a inhibition rate of 30.7％,28.2％,and 55.8％,respectively.The coefficient of drug interaction (CDI) of docetaxel and Ad-NDRG2 was 0.89.Conclusions Ad-NDRG2 can enhance the growth suppression and apoptosis induced by docetaxel in synergistic way in vitro and in vivo.It demonstrated the great potential of Ad-NDRG2 in the treatment of androgen-independent prostate carcinoma.