沉默MAD2基因对结肠癌细胞的影响  

Silencing of MAD2 gene on effect of human colon cancer cells line

作　　者：秦宣[1,2] 王常刚[1,2] 季晓频[1] 张晓健[1,2] 陈雪华[2] 王振雷[1,2] 邓漾[1] 赵任[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院外科,上海200025 [2]上海交通大学医学院附属瑞金医院上海消化外科研究所,上海200025

出　　处：《外科理论与实践》2013年第3期242-246,共5页Journal of Surgery Concepts & Practice

摘　　要：目的:研究沉默MAD2基因对结肠癌细胞的影响。方法 :采用RT-PCR和Western印迹法筛选MAD2基因在7种结肠癌细胞中的高表达细胞株,并用siRNA瞬时转染以降低MAD2表达,随后采用CCK-8法检测结肠癌细胞增殖能力。流式细胞仪检测细胞凋亡和周期,transwell实验评估转染后结肠癌细胞迁移能力的改变。结果:SW1116细胞在mRNA和蛋白水平均呈现高表达MAD2,转染靶向MAD2的siRNA可显著降低MAD2在SW1116细胞中的表达。下调MAD2后,SW1116细胞的增殖能力较对照组有明显下降(P<0.05)。干扰siRNA组细胞凋亡比例较对照组增加,而细胞周期未见统计学差异。transwell实验中,siRNA干扰组穿膜细胞数为(45±2)个,显著低于阴性对照组[(185±5)个]和空白对照组[(195±5)个](P<0.01)。结论:SW1116高表达MAD2基因,沉默MAD2基因显著降低肠癌细胞增殖,诱导凋亡,并抑制细胞迁移。MAD2基因很可能成为新的预防结肠癌转移的目的基因。Objective To investigate the influence of MAD2 gene silencing on the human colon cancer cells. Methods RT-PCR and Western blot were applied in seven colon cancer cell lines to select the cell which highly expresses MAD2. siRNA targeting MAD2 was transiently transfected to down-regulate MAD2 expression. Cell counting kit-8 was used to detect the influence of MAD2 on the regulation on proliferation. Apoptosis and cell cycle were measured by flow eytometry （FCM）. Changes in invasion ability of the selected cell line were evaluated by transwell assay. Results SW1116 had high expression of MAD2 both at mRNA and protein level. MAD2 targeting siRNA transfection in SWlll6 cells significantly reduces MAD2 expression. Compared with the control groups, the proliferation of siRNA group was significantly suppressed （P〈0.05）. FCM revealed that the percentage of apoptosis cells was higher in the siRNA group than in the control groups. No difference was present in the cell cycle. In transwell assay, the number of cells at the bottom of chambers in siRNA group was （45 ±2）, which was lower than that in either the negative control group （185±5） or blank control group （195±5）（P〈0.01）. Conclusions SWII16 highly expresses MAD2. Silencing of MAD2 can inhibit significantly cell proliferation, induce apoptosis and suppress migration. MAD2 may represent as a novel target gene for therapeutic intervention to prevent colon cancer metastasis.

关 键 词：结肠癌 细胞增殖 细胞迁徙 MAD2基因 

分 类 号：R735.35[医药卫生—肿瘤]