The Effect Of The Lentivirus-mediated RNA Interference Targeting Survivin Gene On The Growth Of PC-3 Cells In Vitro And Vivo

PART oneConstruction of PC-3 cell lines with survivin gene stableinterference by lentivirusObjective Construction of survivin gene interference PC-3 cell lines.Methods The PC-3 cell was infected by the lentivirus containing the shRNA targingsurvivin gene, the mRNA and protein level of survivin was detected by RT-PCR andflowcytotechnology. The result was compared with the control group.Results The mRNA and protein level of survivin of PC-3 cell infected with survivinshRNA lentivirus[(5.89±0.12)%,(11.82±0.32)%] was significantly lower than thenegative control group[(55.17±0.87)%,(95.70±1.56)%] and the blank control PC-3cell[(58.21±0.68)%,(95.90±1.78)%].Conclusion The PC-3 cell line with survivin gene stable interference wassuccessfully constructed through lentivirus vector.PART twoThe effect of RNA interference targeting survivin gene on theapoptosis and growth of PC-3 cellsObjective To study the effect of stable interference of survivin gene on theapoptosis and growth of PC-3 cell in vitro.Methods The caspase-3 protein,the level of apoptosis and the cell growth wereexamined by western-blot,flowcytotechnology and cell count respectively in thePC-3,PC-3/c and PC-3/i cells.Results The relative mount of caspase-3 protein was: PC-3 group (0.210±0.021),PC-3/c group (0.240±0.019),PC-3/i group (0.574±0.041); the apoptosis ratio was:PC-3 group (2.3%),PC-3/c group(2.6%),PC-3/i group(22.5%); the cell growth ofPC-3/i was inhibited obviously compared with PC-3 and PC-3/c cells.Conclusion The apoptosis level of PC-3 cell increased and cell growth was inhibited in vitro when the survivin gene interfered by the lentivirus vector.PART threeThe effect of RNA interference targeting survivin gene on theapoptosis and growth of PC-3 xenograft in nude miceObjective To study he effect of RNA interference targeting survivin gene on theapoptosis and growth of PC-3 xenograft in nude mice.Methods The PC-3 cells were injected into the flank of the nude mice to establishthe exnograft, the volumes of the tumor were measured twice a week after 1 week.All the mices were sacrificed after 7 weeks of implantation, the weights of the tumorwere recorded and the TUNEL was done to study the level of tumor apopstosis.Results The volumes and weights of each group were PC-3/i group: 236.1±51.2mm~3,0.21±0.04g; PC-3 group: 932.8±113.2 mm~3,0.75±0.12g; PC-3/c group:930.9±115.2 mm~3,0.71±0.11g. The percentage of TUNEL positive cells of eachgroup was PC-3/i group: 38.4±9.8%; PC-3/c group: 14.5±1.1%; PC-3 group:12.3±1.2%.Conclusion The growth of the PC-3 xenograft was significantly inhibited by theRNAi targeting survivin gene.