Inhibitory effect of TRPV6 silencing on prostate cancer cell line LNCaP in vitro.
- Author:
Xiao-zhi ZHAO
1
;
Hong-qian GUO
;
Guang-xiang LIU
;
Chang-wei JI
;
Shi-wei ZHANG
;
Tie-shi LIU
;
Wei-dong GAN
;
Xiao-gong LI
Author Information
- Publication Type:Journal Article
- MeSH: Apoptosis; Calcium Channels; genetics; Cell Cycle; drug effects; Cell Line, Tumor; Cell Proliferation; drug effects; Gene Silencing; Humans; Male; Prostatic Neoplasms; genetics; pathology; RNA, Small Interfering; genetics; pharmacology; TRPV Cation Channels; genetics; Transfection
- From: National Journal of Andrology 2010;16(5):423-427
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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.
METHODSWe 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.
RESULTSBoth 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).
CONCLUSIONTRPV6-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.