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Published on: May 21, 2019
[Inhibitory effect of TRPV6 silencing on prostate cancer cell line LNCaP in vitro]
Xiao-zhi Zhao1, Hong-qian Guo, Guang-xiang Liu
1Department of Urology, Drum Tower Hospital Affiliated to Nanjing University School Medicine, Nanjing, Jiangsu 210008, China.
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.
Insights
Small interfering RNA (siRNA) targeting the TRPV6 gene effectively reduced TRPV6 mRNA levels in prostate cancer cells. This gene silencing inhibited cell proliferation, arrested the cell cycle, and induced apoptosis in LNCaP cells.
Area of Science:
- Molecular Biology
- Cancer Research
- Gene Silencing
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