| Objiective To explore the expression of chicken anemia virus(CAV)VP3 Gene in prostate cancer cell lines PC-3;and then observe apoptosis inducing effect of CAV VP3 gene singlly or combination Docetaxel on PC-3 cells.METHODS The recombinant erkaryotic expression vector PcDNA3-VP3 was constructed.Then,using Lipofectamine TM2000 mediated transfection in vitro,the positive recombinant containing VP3 gene was transfected into human prostate cancer cell lines PC-3. RT-PCR was performed to detect the expression condition of VP3 gene.after that,MethoCell morphology,HE staining,MTT,flow cytometer method were used to observe the effect of PcDNA3-VP3 or(and)combination 10-8mol/L,10-7mol/L,10-6mol/L Docetaxel on prostate cancer cell line PC-3 in vitro.RESULTS We constructed the recombinant plasmids succededfully,which was corroborated by analyse of gene sequence respectively.The results of RT-PCR suggested that VP3 gene was expressed in PC-3 cells.The typical morphological changes of apoptosis such as cell shrinkage,chromatin condensation,nuclear fragmentation and apoptotic body were observed by both light or electron microscope.The proliferation of PC-3 cells of VP3 singlly done or 10-7mol/L,10-6mol/L Docetaxel singlly done and the combination VP3 and 10-8mol/L,10-7mol/L,10-6mol/L Docetaxel done which was detected by MTT method are all decrease distinctively(P＜0.01).But 10-8mol/L Docetaxel done singly was detected by MTT method are not decreased distinctively.The apoptotic rates detected by TUNAL method of VP3 gene singlly done or the combination VP3 and 10-8mol/L,10-7mol/L Docetaxel done after transfection for 24,48,72 hours were(20.31±1.96)%,(41.50±1.03)%,(50.03±3.00)%(P＜0.01);(31.10±0.59)%,(53.10±0.77)%,(68.90±2.66)%(P＜0.01);(40.01±0.53)%,(62.23±0.74)%,(75.20±0.53)%(P＜0.01)。The apoptotic rates were all higher than that in control group(P＜0.01).The combination one team(10-8mol/L Docetaxel)was also higher than the VP3 singlly done(P＜0.01).CONCLUSION The expression of chicken anemia virus VP3 gene can induce early and high efficiency apoptosis in human prostate cancer lines PC-3.The combination of VP3 gene and the density of exceeding 10-8mol/L Docetaxel can accrete the growth suppression and induce apoptosis of PC-3 cells in vitro.These results will provide a foundation for the future work such as the combination of gene therapy and chemotherapy of the HIPC.And the density of Docetaxel is probably decreased when combination is done,which is useful in clinical. |
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