文章摘要
常晓嫒,朱云霞,韩 晓,程 光.microRNA-24通过下调Cdk4和CyclinD3抑制胰岛β细胞增殖[J].南京医科大学学报,2016,(12):1413~1417
microRNA-24通过下调Cdk4和CyclinD3抑制胰岛β细胞增殖
microRNA-24 inhibits islet β-cell proliferation by decreasing Cdk4 and CyclinD3 protein levels
投稿时间:2016-07-19  
DOI:10.7655/NYDXBNS20161201
中文关键词: microRNA  胰岛β细胞  增殖抑制
英文关键词: microRNA  islet β-cell  proliferation inhibition
基金项目:
作者单位
常晓嫒 南京医科大学江苏省人类功能基因组学重点实验室,江苏 南京 211166 
朱云霞 南京医科大学江苏省人类功能基因组学重点实验室,江苏 南京 211166 
韩 晓 南京医科大学江苏省人类功能基因组学重点实验室,江苏 南京 211166 
程 光 南京绿叶制药有限公司长效和靶向制剂国家重点实验室,江苏 南京 210061 
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中文摘要:
      目的:通过共同过表达Cdk4?CyclinD3与microRNA-24,观察miR-24所引起的胰岛β细胞增殖抑制是否得到逆转,从而判定Cdk4?CyclinD3是否介导了这种抑制作用?方法:在胰岛β细胞系MIN6细胞中过表达miR-24,运用WST-1法检测细胞活力,借助于流式细胞术检测细胞周期,Hoechst33342染色和BrdU标记检测细胞增殖凋亡情况?Western blot检测Cdk4?CyclinD3蛋白水平?构建cyclinD3-pCMV5-myc,cdk4-pCMV5-myc重组质粒,与pre-miR-24?pre-Neg miRNA precursors共转染MIN6细胞,BrdU标记检测细胞增殖?结果:过表达miR-24引起MIN6细胞活力降低,细胞周期G2期阻滞,细胞增殖受到抑制而并没有显著的凋亡;过表达miR-24降低了Cdk4?CyclinD3蛋白水平;而Cdk4?CyclinD3的过表达逆转了miR-24引起的胰岛β细胞增殖抑制?结论:miR-24通过降低Cdk4?CyclinD3蛋白水平抑制胰岛β细胞增殖,该作用可被Cdk4?CyclinD3蛋白的过表达逆转?
英文摘要:
      Objective:To explore whether Cdk4 and CyclinD3 are involved in the inhibition of β-cell proliferation caused by miRNA-24. Methods:miRNA-24(miR-24)was highly expressed in MIN6 cells. Cell viability was determined using WST-1 assays. Cells were collected for flow cytometry analysis to test cell cycle distribution. Protein levels of Cdk4 and CyclinD3 were analyzed by Western blot. CyclinD3-pCMV5-myc and cdk4-pCMV5-myc recombinant plasmid were constructed and transfected into MIN6 cells together with pre-miR-24 or pre-Neg miRNA precursors. Cell proliferation was measured by the BrdU labeling. Results:MIN6 cells proliferation was reduced because of transfected miR-24,mainly due to G2 phase arrest. No changes of the level of apoptotic cells were detected. Cdk4 and CyclinD3 were downregulated by overexpressed miR-24. Overexpressed cdk4 and cyclinD3 reversed the β-cell proliferation inhibition. Conclusion:miR-24 inhibits β-cell proliferation by decreasing Cdk4 and CyclinD3 protein levels. The inhibition may be reversed by overexpressed cdk4 and cyclinD3.
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