The Influence Of Survivin Gene On MiRNAs Expression In Prostate Cancer PC-3 Cells

PartⅠThe effect of survivin gene inhibition mediated by RNA interference in prostate cancer PC-3 cell lineObjective:To examine the effect of survivin gene inhibition mediated by RNA interference in prostate cancer PC-3 cell lines.Methods:A recombinant adenovirus (pGsadeno-sur shRNA) with survivin short hairpin RNA(shRNA) was constructed and used to transfect PC-3 cells. MTT assay and flow cytometry were used to monitor the proliferation activity and apoptosis rate of PC-3 cell after transfection respectively. RT-PCR and western blot technologies were used to detect the expression of gene and protein of survivin.Results:The recombinant adenovirus was successfully constructed, transfected and duplicated in PC-3 cells. Compared with the control group, the experimental group shows a noticeable inhibition of PC-3 cells proliferation and increased apoptosis at different time. Ninety six hours after transfection, the expression of survivin mRNA and protein decreased by (72.23±1.11)% and (71.75±2.64)%, respectively,and the apoptosis rate reached (14.43±0.57)%.In contrast to the negative control group and the blank group, the deviation is statistically significant.Conclusion:The Adenovirus vector mediated by siRNA could inhibit survivin effectively in PC-3 cells. The combination of RNAi with Adenovirus carrier system to inhibit survivin could create a new modus to cure the androgen-independent prostate cancer. PartⅡExperimental studies of miRNAs expression profile in survivin-inhibited PC-3 cellsObjective:To detect the miRNAs expression profile and predict correlation between survivin and related miRNAs in surviving-inhibited PC-3 cell lines.Method : A recombinant adenovirus (pGsadeno-sur shRNA) with survivin short hairpin RNA(shRNA) was constructed and used to transfect PC-3 cells. 96 hours after transfection, the cells were then collected with small RNAs isolated from all groups of PC-3 cells and the expression profiles were analyzed with microarray technologies.Results : Altogether 650 miRNAs signals were detected in normal androgen independent prostatic carcinoma cells, of which 9 miRNAs have a signal level of over 1000. Compared with the blank group, the experimental group has 75 miRNAs exhibiting statistically significant deviations, 42 with significant up-regulation and 33 with significant down-regulation (p<0.01). Compared with the negative control group, the experimental group has 27 miRNAs expression displaying statistically significant difference(p<0.01), with 22 cases of up-regulation and 5 cases of down-regulation. In contrast to the blank and negative control, the experimental group has 15 miRNAs with up-regulated expression, and 1 down-regulated expression (p<0.01).Conclusion: The inhibition of the expression of the survivin in PC-3 cell lines has a great effect on the miRNAs expression profile. The findings open a new perspective for uncovering related miRNAs'mechanism in PC-3 cell lines.