c-Myc promotes tubular cell apoptosis in ischemia-reperfusion-induced renal injury by negatively regulating c-FLIP and enhancing FasL/Fas-mediated apoptosis pathway
c-Myc promotes tubular cell apoptosis in ischemia-reperfusion-induced renal injury by negatively regulating c-FLIP and enhancing FasL/Fas-mediated apoptosis pathway
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c-Myc 通过负向调节 c-FLIP 和增强 FasL/Fas 介导的凋亡途径促进缺血再灌注肾损伤中肾小管细胞凋亡
DOI：
10.1038/s41401-018-0201-9
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通讯作者：
Lu L
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作者：
Xu D;Wang B;Chen PP;Wang YZ;Miao NJ;Yin F;Cheng Q;Zhou ZL;Xie HY;Zhou L;Liu J;Wang XX;Xue H;Zhang W;Lu L
c-Myc 在细胞增殖、分化和细胞凋亡中发挥重要作用。 FasL/Fas 通路是细胞凋亡的关键调节因子。本研究旨在探讨 c-Myc 对缺血再灌注 (I/R) 诱导的肾损伤中 FasL/Fas 通路的影响。大鼠双侧肾缺血 60 分钟，再灌注 24 或 48 小时。 NRK-52E 细胞用缺氧复氧 (H/R) 或 FasL 处理。使用免疫组织化学来鉴定c-Myc的分布。通过TUNEL染色评估细胞凋亡。 Ad-c-Myc 和重组 pcDAN 3.0 分别用于过表达 c-Myc 和 c-FLIP。采用 ChIP 测定和荧光素酶测定来检测 c-Myc toc-FLIPpromoter 的结合。 I/R大鼠中c-Myc显着升高，主要定位于肾小管上皮细胞；同时，c-FLIP减少，cleaved caspase-8、cleaved caspase-3和TUNEL阳性染色细胞增多。用c-Myc抑制剂10058-F4治疗I/R大鼠可显着减弱I/R大鼠中c-FLIP的减少、cleaved caspase-8、cleaved caspase-3、TUNEL阳性细胞、Scr和BUN的增加。在NRK-52E细胞中，缺氧和复氧诱导c-Myc增加和c-FLIP减少。 ChIP 和荧光素酶测定结果表明 c-Myc 与 c-FLIP 基因的启动子区域结合。 c-Myc 的过表达显着降低了 c-FLIP。 c-FLIP 的过表达抑制了 FasL 诱导的 caspase-8 和 caspase-3 裂解的增加。数据表明，I/R 大鼠肾脏中的 c-Myc 增加，并负向调节 c-FLIP 的表达，然后增强 FasL 诱导的 I/R 应激中的细胞凋亡。
c-Myc plays an important role in cell proliferation, differentiation, and cell apoptosis. FasL/Fas pathway is a key regulator of cell apoptosis. This study was aimed to investigate the effects of c-Myc on the FasL/Fas pathway in ischemia-reperfusion (I/R)-induced renal injury. Rats were objected to bilateral renal ischemia for 60 min and reperfused for 24 or 48 h. NRK-52E cells were treated with hypoxia-reoxygenation (H/R) or FasL. Immunohistochemistry was used to identify the distribution of c-Myc. Cell apoptosis was assessed by TUNEL staining. Ad-c-Myc and recombinant pcDAN 3.0 were used to overexpress c-Myc and c-FLIP, respectively. ChIP assay and luciferase assay were used to detect the binding of c-Myc toc-FLIPpromoter. In I/R rats, c-Myc was increased significantly and mainly located in renal tubular epithelial cells; meanwhile, c-FLIP was decreased, cleaved caspase-8, cleaved caspase-3 and TUNEL-positive staining cells were increased. Treatment of I/R rats with c-Myc inhibitor 10058-F4 significantly attenuated the decrease in c-FLIP, the increase in cleaved caspase-8, cleaved caspase-3, TUNEL-positive cells, Scr and BUN in I/R rats. In NRK-52E cells, hypoxia and reoxygen induced the increase in c-Myc and decrease in c-FLIP. ChIP and luciferase assay results indicated that c-Myc binds to the promoter region ofc-FLIPgene. Overexpression of c-Myc markedly decreased c-FLIP. Overexpression of c-FLIP inhibited the increase in cleaved caspase-8 and caspase-3 induced by FasL. Data indicated that c-Myc is increased in kidneys of I/R rats and negatively regulates the expression of c-FLIP, then enhanced FasL-induced cell apoptosis in I/R stress.