Activation of Rip1 promotes necroptosis in LNCaP-AI cells via inhibiting SHARPIN
10.3969/j.issn.1000-4718.2016.07.010
- VernacularTitle:抑制SHARPIN激活Rip1促进LNCaP－AI细胞坏死性凋亡
- Author:
Ganping WANG
;
Hai HUANG
;
Xianju CHEN
;
Yiming LAI
;
Chunhao LIN
;
Lexiang ZENG
;
Yi CAO
;
Yiming ZHANG
;
Yongsheng YU
;
Zhenghui GUO
- Publication Type:Journal Article
- Keywords:
Castration-resistant prostate cancer;
LNCaP-AI cells;
Rip1;
SHARPIN;
Necroptosis
- From:
Chinese Journal of Pathophysiology
2016;32(7):1214-1220
- CountryChina
- Language:Chinese
-
Abstract:
[ ABSTRACT] AIM:To explore the role of SHARPIN in regulation of Rip1 in castration-resistant prostate cancer LNCaP-AI cells.METHODS:The LNCaP-AI cells were treated with TNF-α+Z-VAD ( an inhibitor of pan-caspase) to activate necroptosis, which were compared to the cells treated with TNF-α+Z-VAD+Nec-1 ( an inhibitor of Rip1 ) .A blank group and a TNF-α-treated group were set up as controls.The cell viability in each group was measured by MTS as-say.In addition, SHARPIN was knocked down by siRNA, and the inhibitory efficiency was evaluated by RT-qPCR.The expression of Rip1 at mRNA and protein levels after knocking down SHARPIN was determined by RT-qPCR and Western blot to explore the underlying mechanism of regulatory network of necroptosis in prostate cancer.RESULTS: Compared with blank control group and TNF-α-treated group, the viability of LNCaP-AI cells treated with TNF-α+Z-VAD decreased by 28%(P<0.05).After treated with TNF-α+Z-VAD+Nec-1, the LNCaP-AI cells showed no significant difference in the viability compared with blank control and TNF-α-treated groups.Taken together, necroptosis may be an important way of cell death in LNCaP-AI cells.Besides, the expression of Rip1 at protein level was up-regulated following the inhibition of SHARPIN using siRNA, indicating that down-regulation of SHARPIN enhanced necroptosis via activating Rip1 in
LNCaP-AI cells.CONCLUSION:Necroptosis is an important way of cell death .Inhibition of oncogenic factor SHARPIN enhances necroptosis via activating Rip1 in LNCaP-AI cells.
