siRNA抑制EPCAM基因表达对结肠癌HCT116细胞株增殖和侵袭的影响  

Effects of EPCM gene expression downregulation by siRNA on the proliferation and invasion of colon cancer cell line HCT116

作　　者：张泽栋[1] 周逢强[2] 徐宏[2] 刘磊[2] 齐艳美[3] 

机构地区：[1]滨州医学院,山东滨州256600 [2]滨州市人民医院普外科,山东滨州256600 [3]滨州市人民医院消化内科,山东滨州256600

出　　处：《中国现代普通外科进展》2015年第5期348-351,360,共5页Chinese Journal of Current Advances in General Surgery

摘　　要：目的:利用si RNA沉默EPCAM基因,观察对结肠癌HCT116细胞增殖和侵袭能力的影响。方法:将针对EPCAM基因不同位点的3个si RNA(分别命名为EPCAM-si RNA-1、EPCAM-si RNA-2、EPCAM-si RNA-3)以及阴性对照组和空白对照组分别转染结肠癌HCT116细胞,利用RT-PCR和Western blot方法检测各组细胞EPCAM m RNA和蛋白的表达。选取对目的基因沉默效果最强的一组si RNA转染结肠癌细胞,MTT法检测转染前后细胞的增殖活性,transwell实验检测转染前后细胞的侵袭能力。结果:3条EPCAM si RNA均能有效抑制EPCAM基因的表达,其中EPCAM-si RNA-1抑制效果最强,与空白对照组相比,EPCAM m RNA和蛋白表达量分别降低59.6%和54.5%。MTT结果表明EPCAM-si RNA-1组HCT116细胞生长速度明显低于阴性对照组和空白对照组(P<0.05),Transwell实验结果表明实验组细胞侵袭能力明显低于两个对照组(P<0.05)。结论:EPCAM si RNA可以有效抑制结肠癌HCT116细胞EPCAM的表达,可以明显降低HCT116细胞的增殖和侵袭能力。Objective： To observe the effect on proliferation and invasion in colon cancer cell HCT116 after inhibition of EPCAM expressing by si RNA. Methods： Three EPCAM si RNA sequences and one negative control si RNA sequence were designed and synthesized,and then transfected into HCT116 cells.The most effective EPCAM- si RNA was screened by real time PCR and Western blotting.Then the target si RNA was used to test the proliferative and invasive ability of colon cancer cell HCT116. Result： All of the three EPCAM- si RNA could effectively restrain EPCAM gene expression, EPCAM- si RNA- 1 was the most effective,compared with the control group, EPCAM m RNA and protein expression quantity reduced by 59.6% and 54.5% respectively. After transfection, EPCAM- si RNA- 1 group showed an obvious proliferation arrest compared with the negative control group, and the invision ability has also been restrained obviously in group EPCAM- si RNA- 1. Conclusion： EPCAM si RNA can effectively inhibit the expression of EPCAM gene in colon cancer line HCT116, and downregulation of EPCAM expression can significantly depress the proliferation and invasion of colon cancer line HCT116.

关 键 词：RNA干扰 结肠肿瘤 上皮细胞黏附分子 增殖 侵袭 

分 类 号：R735.3[医药卫生—肿瘤]