微小RNA-29b-1＊和微小RNA-29c对膀胱癌T24细胞增殖的影响  

Effect of microRNA.29b-1 ＊ and microRNA-29c on the proliferation of bladder cancer cell line T24

作　　者：张庆玲[1] 徐锋[1] 程文[1] 刑潇潇 张征宇[1] 葛京平[1] 

机构地区：[1]南京军区南京总医院泌尿外科,210002

出　　处：《中华实验外科杂志》2013年第10期2129-2132,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30772278）

摘　　要：目的 观察微小RNA（miR）-29b-1＊和miR-29c对膀胱癌T24细胞的活性和增殖的影响.方法 构建携带目标miRNAs干扰基因的慢病毒（pGCsil-H1-CMV-GFP）载体、阳性克隆的聚合酶链反应（PCR）鉴定.用293T细胞包装慢病毒形成慢病毒重组体,并用孔稀释法测定滴度.T24细胞中转染慢病毒重组体及转染效率测定.用噻唑蓝（MTT）比色法对4组处于对数生长期的实验组[空细胞对照组、转染阴性对照慢病毒细胞组、转染miR-29b-1＊基因RNA干扰（RNAi）慢病毒细胞组、转染miR-29c基因RNAi慢病毒细胞组]进行连续5d的细胞增殖分析.结果 实时定量PCR（Real-time PCR）验证结果显示：T24细胞中miR-29b-1 ＊和miR-29c表达水平比较于空细胞组分别为3.47和4.40；MTT比色法显示,转染阴性对照慢病毒细胞组与空细胞对照组细胞比较,增殖受到轻度抑制；而转染miR-29b-1＊、miR-29c基因RNAi慢病毒细胞组与染阴性对照慢病毒细胞组和空细胞对照组细胞比较,增殖均受到明显抑制,前者抑制效果更明显.结论 细胞增殖MTT法分析显示通过RNAi降低miR-29b-1＊、miR-29c的表达均能引起T24细胞的增殖抑制.Objective To investigate the effect of microRNA （miR）-29b-1 ＊ and miR-29c on the proliferation of bladder cancer cell line T24.Methods Transfection Structuring lentivirus （pGCsil-H1-CMV-GFP） vector with interfering RNA of target miRNAs （miR-29b-1 ＊,miR-29c）and indentified by polymerase chain reaction （PCR）.Titer of mature lentivirus vector recombinant was measured by dilution plate method after packed in 293T cell.Then transfect lentivirus vector recombinants to T24 cell.Cell proliferation analysis with methyl thiazol tetrazolium （MTT） cell proliferation was evaluated by the trypan blue method with MTT in four groups （blank control,nagetive control-transfectants,miR-29b-1 ＊-RNA interference （RNAi） transfected,miR-29c-RNAi transfected）.Results miR-29b-1 ＊ and miR-29c were abnormally expressed in T24 cell content with our previous study on BUC specimens with a fold of 3.47 and 4.40 respectively according to the data of Real-time PCR result.T24 cell proliferation was both suppressed after miR-29b-1 ＊-RNAi transfected and miR-29c-RNAi transfected compared with negative control and blank control.Conclusion It shows a high efficiency of interfering RNAs of target mRNAs transfection using lentivirus vector for RNAi in our experiment.In this stuy,miR-29b-1 ＊ and miR-29c were considered to be a tumor promotor according their suppression of T24 cell proliferation evaluated by the trypan blue method with MTT after their interfering RNAs transfected.

关 键 词：微小RNA-29b-1＊ 微小RNA-29c 膀胱癌 T24 

分 类 号：R737.14[医药卫生—肿瘤]