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Effects of miRNA-136 overexpression on S100 calcium binding protein A6 contents, proliferation and apoptosis in HCT116 cells
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- Author:
- No author available
- Journal Title:
- Academic Journal of Second Military Medical University
- Issue:
- 9
- DOI:
- 10.3724/SP.J.1008.2012.00940
- Key Word:
- 微RNAs;S100A6蛋白;结直肠肿瘤;HCT116细胞;细胞增殖;细胞凋亡
Abstract： 目的 构建携带微小RNA136(mir136)的真核表达载体并将其转染人结直肠癌细胞,观察外源性mir136表达对人结直肠癌细胞中钙结合蛋白S100A6表达及细胞增殖活性的影响.方法 提取人肾上皮HEK293细胞基因组DNA,PCR扩增包含mir136前体序列的DNA片段并构建重组载体,采用阳离子脂质体法将重组载体转染人结直肠癌HCT116细胞.转染后48h,采用实时定量PCR法检测细胞内mir136含量变化,以蛋白质印迹法检测细胞中S100A6蛋白的表达；转染后24和48h,采用CCK 8试剂盒检测肿瘤细胞增殖活性；转染后48 h,以双染法检测细胞凋亡情况.结果 成功构建重组mir136真核表达载体并转染HCT116细胞.转染后48 h,重组载体转染组HCT116细胞内mir136相对表达量高于未转染组(P＜0.01),而S100A6蛋白的相对表达量则低于未转染组(P＜0.01).同时,重组载体转染组HCT116细胞增殖活性较未转染对照组降低(P＜0.05),而细胞凋亡则较未转染对照组增加(P＜0.01).结论 Mir136可能通过抑制S100A6基因表达而抑制结直肠癌细胞增殖活性,同时引起细胞的凋亡.
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