miR-143-5p对镉致肾细胞凋亡的调控作用及其机制

发布时间：2018-04-22 04:31

本文选题：镉 + 凋亡　；参考：《南京医科大学学报(自然科学版)》2015年04期

【摘要】：目的 :探讨mi R-143-5p对镉诱导LLC-PK1细胞凋亡的调控作用及其机制。方法 :通过基因芯片技术筛选出由镉引起的差异表达mi RNAs,并用实时定量PCR方法验证基因芯片结果的可靠性;应用Lipofectamine 2000瞬时转染mi R-143-5p的模拟物和抑制剂建立mi R-143-5p高表达和低表达的模型,并结合q RT-PCR技术验证转染效果;Hoechst 33258染色和Annexin V-PI双染法检测细胞凋亡;生物信息学分析结合实时定量PCR和Western blot验证mi R-143-5p靶基因的表达;Western blot分析mi R-143-5p对细胞凋亡相关通路的调控作用。结果:镉可增强LLC-PK1细胞的mi R-143-5p表达水平(P0.01);转染mi R-143-5p模拟物或抑制剂后,与对照组(mi R-NC)比较,mi R-143-5p表达明显上调或下调(P0.01);mi R-143-5p的过表达可促进LLC-PK1细胞凋亡(P0.01);mi R-143-5p在AKT3的m RNA和蛋白水平上发挥靶向调控作用;mi R-143-5p过表达能抑制pAkt和p-Bad蛋白表达,促进caspase-9和caspase-3蛋白上调。结论 :mi R-143-5p可能通过作用于靶基因AKT3,并抑制Akt/Bad信号通路,促进镉诱导LLC-PK1细胞的凋亡。
[Abstract]:Aim: to investigate the regulatory effect of mi R-143-5p on cadmium induced apoptosis in LLC-PK1 cells and its mechanism. Methods: the differentially expressed mi RNAs-induced by cadmium was screened by gene chip technique, and the reliability of the results was verified by real-time quantitative PCR. The model of high expression and low expression of mi R-143-5p was established by transient transfection of Lipofectamine 2000 into the mimic and inhibitor of mi R-143-5p. The effect of transfection was verified by Q RT-PCR technique. Apoptosis was detected by Annexin V-PI double staining and Hoechst 33258 staining. Bioinformatics analysis combined with real-time quantitative PCR and Western blot was used to verify the expression of mi R-143-5p target gene. Western blot was used to analyze the regulatory effect of mi R-143-5p on apoptosis-related pathway. Results: cadmium could enhance the expression level of mi R-143-5p in LLC-PK1 cells. Compared with the control group, the overexpression of R- 143-5p significantly up-regulated or down-regulated the overexpression of P0.01OMR-143-5p in LLC-PK1 cells. The overexpression of R- 143-5p could promote the apoptosis of LLC-PK1 cells. P0.01nmR-143-5p could inhibit the expression of pAkt and p-Bad proteins at the level of m RNA and protein of AKT3, and the overexpression of R- 143-5p could inhibit the expression of pAkt and p-Bad protein, and the overexpression of R- 143-5p could inhibit the expression of pAkt and p-Bad. Promote the upregulation of caspase-9 and caspase-3 protein. Conclusion it is suggested that the apoptosis of LLC-PK1 cells induced by cadmium may be promoted by the action of the target gene AKT3 and the inhibition of Akt/Bad signaling pathway by the action of 1: mi R-143-5p.
【作者单位】：南京医科大学公共卫生学院营养与食品卫生学系;
【基金】：江苏高校优势学科建设工程资助项目(2011)
【分类号】：R114

【共引文献】