摘要
目的探究siRNA靶向抑制Tspan-1对结直肠癌细胞增殖和侵袭能力的影响。方法实验组针对Tspan-1基因2个不同位点进行siRNA沉默,分别命名为Tspan-1-siRNA-1、Tspan-1-siRNA-2。采取完全剔除Tspan-1基因的作为阴性对照组,同时选取空白对照组。Wester blot方法检测各组的Tspan-1mRNA和蛋白的表达情况,选取基因沉默效果最强的一组进行结肠癌细胞HCT116细胞的转染,分别对转染前后细胞的增殖能力和侵袭能力进行检测,对siRNA靶向抑制Tspan-1能力进行评价和分析。结果相比于空白对照组,实验组各组细胞mRNA均出现了不同程度的下降,其中Tspan-1-siRNA-1、Tspan-1-siRNA-2,两组分别相比对照组下降61. 2%,57. 2%。随着转染时间的增加,实验组细胞增殖的速度明显放缓,其中第2、3、4、5天吸光度值要明显低于阴性对照组和空白对照组,差异有显著性(P <0. 05);同时两个对照组之间的吸光度值无显著差异(P> 0. 05);经过转染24 h后,实验组转移到膜上的细胞数要显著少于对照组,而其余两组无显著差异(P> 0. 05)。结论 Tspan-1基因经过siRNA沉默之后,能够有效降低结肠癌HCT116细胞的增殖和侵袭能力,siRNA靶向抑制Tspan-1基因可以作为治疗结直肠癌重要的的分子手段之一。
Objective To investigate the effect of siRNA targeting inhibition of Tspan-1 on proliferation and invasion of colorectal cancer cells.Methods Two loci of siRNA silencing of Tspan-1 were named as Tspan-1-siRNA-1 and Tspan-1-siRNA-2.The negative control group were without Tspan-1 gene.The blank control group were with untreated Tspan-1.Tspan-1 mRNA and protein were tested by Western blotting to select the best silenced gene for HCT116 cell transfection.Cell proliferation and invasion before and after transfection were detected and inhibition efficacy were evaluated.Results Tspan-1-siRNA-1 and Tspan-1-siRNA-2 decreased Tspan-1 mRNA by 61.2%and 57.2%respectively compared with blank control group.With the increase of transfection time,cells in experimental group showed lowered proliferation rate compared with two control groups.Cells in experimental group metastasized less than two control groups(P<0.05).Conclusion Tspan-1 gene silenced by siRNA can effectively inhibit proliferation and invasion of colon cancer cells and might be molecularly effective against colorectal cancer.
作者
史小玉
吴帮卫
商海涛
SHI Xiao-yu;WU Bang-wei;SHANG Hai-tao(Department of Digestive System, Cadre Ward of General Hospital of PLA Navy, Beijing 100048, China;Department of Cardiology, Huashan Hospital Affiliated to Fudan University, Shanghai 200040, China;Department of Digestive System, Binzhou Medical University Hospital, Binzhou Shandong 256603, China)
出处
《临床和实验医学杂志》
2019年第1期28-31,共4页
Journal of Clinical and Experimental Medicine
