| 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 assay. 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. |
