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目的探讨miR-34a对高糖环境中HK-2细胞增殖作用的影响。方法以miR-34a模拟物和miR-34a抑制剂分别转染HK-2细胞,定量PCR检测miR-34a的表达变化,MTT检测细胞增殖活性;Western印迹检测周期相关蛋白CyclinD1和CyclinE的表达变化。生物信息学分析miR-34a的靶基因,对其中的增殖相关转录因子E2F3进行双荧光素酶验证。结果 miR-34a模拟物组miR-34a表达显著上调(P<0.05),miR-34a抑制剂组miR-34a显著下调(P<0.05)。与对照组相比,miR-34a模拟物显著抑制高糖环境中HK-2细胞的增殖作用(P<0.05),CyclinD1和CyclinE表达下调(P<0.05),miR-34a抑制剂组显著上调高糖环境中HK-2细胞的增殖作用,CyclinD1和CyclinE表达上调(P<0.05)。TargetScan在线分析软件显示,增殖相关转录因子E2F3可能是miR-34a潜在的靶基因,双荧光素酶分析结果显示E2F3是miR-34a的直接靶基因。结论 miR-34a可以调控高糖环境中HK-2的细胞增殖,其机制可能是通过直接抑制E2F3基因的表达而实现。
Objective To investigate the effect of miR-34a on the proliferation of HK-2 cells in high glucose environment. Methods HK-2 cells were transfected with miR-34a mimics and miR-34a inhibitor respectively. The expression of miR-34a was detected by quantitative PCR. The proliferation of cells was detected by MTT assay. The expressions of CyclinD1 and CyclinE were detected by Western blotting. Bioinformatics analysis of miR-34a target gene, which proliferation-related transcription factor E2F3 dual-luciferase validation. Results miR-34a expression was significantly up-regulated in miR-34a mimics (P <0.05), miR-34a was significantly down-regulated in miR-34a inhibitor group (P <0.05). Compared with the control group, miR-34a mimics significantly inhibited the proliferation of HK-2 cells in high glucose (P <0.05), while the expression of CyclinD1 and CyclinE were down-regulated (P <0.05) The proliferation of HK-2 cells in the glucose environment, the expression of CyclinD1 and CyclinE were up-regulated (P <0.05). TargetScan online analysis software showed that proliferation-related transcription factor E2F3 may be a potential target gene for miR-34a, and dual-luciferase analysis revealed that E2F3 is a direct target of miR-34a. Conclusion miR-34a can regulate the proliferation of HK-2 cells in high glucose environment. The possible mechanism is that miR-34a directly inhibits the expression of E2F3 gene.