抑制SHARPIN激活Rip1促进LNCaP-AI细胞坏死性凋亡

发布时间：2018-04-02 09:54

  本文选题：去势抵抗性前列腺癌　切入点：LNCaP-AI细胞　出处：《中国病理生理杂志》2016年07期

【摘要】：目的:探讨SHARPIN对去势抵抗性前列腺癌细胞LNCaP-AI坏死性凋亡关键因子Rip1的调控作用,以及对细胞坏死性凋亡的影响。方法:将LNCaP-AI细胞分为TNF-α+Z-VAD(caspase抑制剂)处理组与TNF-α+Z-VAD+Nec-1(Rip1抑制剂)处理组,应用MTS检测各组细胞的活力,研究坏死性凋亡机制在诱导LNCaP-AI细胞死亡中的作用。将LNCaP-AI细胞分为阴性对照组和SHARPIN干扰(si-SHARPIN)组,RT-qPCR验证抑制效率,通过免疫荧光等技术进一步探讨SHARPIN调控坏死性凋亡的具体分子机制。结果:与对照组相比,TNF-α+Z-VAD处理组的LNCaP-AI细胞活力下降28%(P0.05),而TNF-α+Z-VAD+Nec-1处理组的细胞在Rip1被抑制后,细胞活力无明显改变。在LNCaP-AI细胞中,通过siRNA抑制SHARPIN表达后,Rip1表达水平上调,同时,LNCaP-AI细胞坏死性凋亡比例升高。结论:LNCaP-AI细胞可通过坏死性凋亡机制诱导死亡,下调SHARPIN可能通过激活Rip1增强LNCaP-AI细胞坏死性凋亡。
[Abstract]:Aim: to investigate the regulatory effect of SHARPIN on necrotizing apoptosis of LNCaP-AI cells and the effect of Rip1 on necrotic apoptosis in Ovariectomized prostate cancer cells.Methods: LNCaP-AI cells were divided into two groups: TNF- 伪 Z-VAD(caspase inhibitor (TNF- 伪 Z-VAD Nec-1(Rip1 inhibitor) and TNF- 伪 Z-VAD Nec-1(Rip1 inhibitor (TNF- 伪 Z-VAD Nec-1(Rip1 inhibitor). MTS was used to detect the cell viability and to study the role of necrotizing apoptosis in inducing the death of LNCaP-AI cells.LNCaP-AI cells were divided into negative control group and SHARPIN interference si-SHARPIN group to verify the inhibition efficiency of RT-qPCR. The specific molecular mechanism of SHARPIN regulating necrotizing apoptosis was further investigated by immunofluorescence technique.In LNCaP-AI cells, siRNA inhibited the expression of SHARPIN and up-regulated the expression of rip1, while the percentage of necrotic apoptosis in LNCaP-AI cells increased.Conclusion death can be induced by necrotic apoptosis in the cell line of 1: LNCaP-AI, and down-regulation of SHARPIN may enhance necrotizing apoptosis of LNCaP-AI cells by activating Rip1.
【作者单位】： 中山大学孙逸仙纪念医院泌尿外科;中山大学孙逸仙纪念医院小儿外科;中山大学孙逸仙纪念医院急诊外科;南方医科大学珠江医院泌尿外科;中山大学附属博济医院泌尿外科;
【基金】：国家自然科学基金资助项目(No.81272807) 广东省科技计划资助项目(No.2014A020212018) 广东省医学研究资助项目(No.A2015113)
【分类号】：R737.25
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本文编号：1699851