Objective: To explore the effects of the TRPV6 gene silencing by small interfering RNA (siRNA) on the proliferation, cell cycle and apoptosis of human prostate cancer LNCaP cells.
Methods: We constructed two siRNA sequences (siTRPV6-1 and siTRPV6-2) targeting the TRPV6 gene and then transfected them into LNCaP cells mediated by liposome. The transcription of TRPV6 mRNA was detected by RT-PCR, and the effects of siRNA on the proliferation, cell cycle and apoptosis of the LNCaP cells were determined by MITT and flow cytometry.
Results: Both siTRPV6-1 and siTRPV6-2 significantly suppressed the expression of TRPV6 mRNA in the LNCaP cells, and the expression was decreased with the extension of time, by 73 and 77% respectively at 72 h after transcription with siTRPV6-1 and siTRPV6-2 as compared with the blank control group (P < 0.01). The proliferation inhibition rates were the highest (34.53 and 29.32%) at 48 h in comparison with 24 and 72 h (P < 0.05). The number of cells was significantly increased in the GO and G1 phases and decreased in the S phase after siTRPV transfection (P < 0.01). The apoptosis rates of LNCaP cells were 14.45 and 12.73% respectively at 48 h after transfected with siTRPV6-1 and siTRPV6-2, significant higher than in the blank control and negative control groups (P < 0.05).
Conclusion: TRPV6-targeted siRNA can effectively inhibit the transcription of TRPV6 mRNA, inhibit the proliferation of LNCaP cells, arrest their cycles in the G0 and G1 phases, and induce their apoptosis.