特异性小干扰RNA靶向沉默KCNN4基因对LoVo细胞生物学影响  

The effects of short interfering RNAs on KCNN4 gene expression and biological behaviors of LoVo cells

作　　者：褚忠华[1] 林显敢[2] 刘璐[1] 伍衡[1] 来伟[1] 

机构地区：[1]中山大学孙逸仙纪念医院胃肠外科,广州510120 [2]中山大学孙逸仙纪念医院肿瘤科,广州510120

出　　处：《中华实验外科杂志》2011年第11期1922-1924,共3页Chinese Journal of Experimental Surgery

基　　金：广东省社会发展计划基金资助项目（200713031513012）;广东省自然科学基金资助项目（9151008901000058、10151008901000071）

摘　　要：目的观察小分子干扰RNA（siRNA）对结肠癌细胞LoVo的KCNN4基因表达及其生物学行为的影响。方法将实验分别空白对照组和实验转染组进行实验。合成针对KCNN4基因的特异性siRNA并转染至结肠癌细胞LoVo中，实时定量逆转录．聚合酶链式反应以及Westernblot分别检测KCNN4mRNA以及蛋白的变化，CCK-8法检测LoVo细胞增殖的变化以及Transwell检测LoVo细胞增殖和侵袭的能力的变化。结果成功合成KCNN4-siRNA并转染人LoVo细胞中。RT—PCR以及Western blot结果提示与对照组比较，KCNN4-siRNA可以明显抑制KCNN4 mRNA以及蛋白的表达，抑制率分别为72．0％、49．6％（P〈0．05）。同时KCNN4-siRNA可以明显抑制LoVo细胞的增殖（P〈0．05）。另外KCNN4-siRNA可以明显抑制LoVo细胞的迁移能力（45．23±3．52比66．42±1．47）和侵袭能力（28．13±1．64比42．78±2．33）。结论KCNN4-siRNA可以有效抑制结肠癌LoVo细胞KCNN4基因的表达，从而有效的抑制肿瘤的增殖、迁移和侵袭。Objective To investigate the effects of small interference RNA interference （siRNA） targeting KCNN4 on biological behaviors of LoVo cell line. Methods LoVo cell lines were divided into two groups： the blank control group, the KCNN4-siRNA group. Synthesized KCNN4-siRNA was transfccted into LoVo cells. Real time quantitative polymerase chain reaction （PCR） and Western blotting were used to detect the KCNN4 mRNA and protein levels. Then the proliferation of LoVo cells was determined by CCK-8 assay. Furthermore, the ability of motility and invasion of LoVo cells were assessed by transwell invasion assay, respectively. Results In the KCNN4-siRNA group, the KCNN4 mRNA and protein levels were inhibited by 72. 0% , 49.6% respectively （ P 〈 0. 05 ）. At the same time, the proliferation of LoVo was also inhibited （ P 〈 0. 05 ）. Furthermore, the ability and invasion of LoVo cells were inhibited significantly （45.23 ±3.52 vs. 66.42 ±1.47;28.13±1.64vs. 42.78 ±2.33, P〈0.05）. Conclusion KC- NN4 could regulate the biological behaviors of LoVo cells line.

关 键 词：结肠癌 小分子干扰RNA KCNN4 

分 类 号：R735.35[医药卫生—肿瘤]