Effect Of Down-regulating ACP1 By RNA Interfering On Prostatic Carcinoma

Objective To construct ACP1 short hairpin RNA eukaryotic expression vector,transfect reconstructed plasmid into prostatic carcinoma cell T3B, and to detect theexpression of ACP1 in T3B and PC-3. Study the biology characteristics changes ofprostatic carcinoma cells after down-regulation of ACP1.Methods Selected the target gene fragment according the open reading frame ofACP1, and designed the short hairpin RNA which synthesized and inserted intopGenesil-1 vector to form cloning plasmid. The recombinant plasmid was identifiedby restriction enzyme digestion and sequencing analysis after E. coli transformation,amplification and purification. Plasmid pGenesil-1/ACP1-shRNA was transfected intoT3B cells with Lipoinfectamin^TM 2000 and then the stably transfected positive cloneswere screened with G418. The mRNA expression of ACP1 was detected by RT-PCRand protein by Western Blot. T3B group, pGenesil-1 group and pGenesil-1/ACP1-shRNA group, PC-3 group cells were detected on growth, cell circle by FCM,capacity of adhesion, migration and invasion.Results Recombinant human ACP1-shRNA eukaryotic expression vector wasconstructed successfully The stably transfected positive clones was successfullyscreened which could express reconstructed ACP1-shRNA stably. The ACP1 mRNAof shRNA transfected group decreased from 0.45±0.03 to 0.09±0.02, compared withcontrol group; ACP1 protein expression dropped from 0.65±0.02 to 0.21±0.03.mRNA was down-regulated 80%, and protein 76%. Compared with pGenesil-1 group,cell growth rate and cell cycle of pGenesil-1/ACP1-shRNA group cells didn't change, the capacity of adhesion, migration and invasion decreased dramatically.Conclusion The expression of ACP1 mRNA and protein was suppressed efficientlyby means of siRNA. Down-regulation of ACP1 could suppress the capacity ofadhesion, migration and invasion in prostatic carcinoma cells, and had no effect oncell proliferation.