摘要
目的 观察小干扰RNA（siRNA）下调假基因Cripto-3（Cr-3）对人大肠癌细胞增殖和侵袭的影响.方法 构建Cr-3 siRNA（S1、S2和S3）,分别以3个siRNA转染大肠癌SW-620细胞后,采用荧光实时定量聚合酶链反应（FQ-PCR）检测筛选出效果最好的siRNA.SW-620细胞分为3组：空白对照组（con-a）、空载质粒对照组（con-b）和Cr-3 siRNA组（siRNA）,其中,siRNA组以Cr-3 siRNA转染处理.用噻唑蓝（MTT）方法检测癌细胞增殖,以Transwell方法检测癌细胞侵袭能力,并作统计学分析.结果 S1、S2和S3均能下调Cr-3 mRNA的表达,有效率分别为83.5％、61.6％和42.8％,说明S1效果最好.以S1转染大肠癌SW620细胞株后,MTT结果显示,72 h con-a、con-b和siRNA组相对增殖率分别为（0.927 ±0.037）％、（0.922 ±0.035）％、（0.663 ±0.018）％ （P ＜0.05）；Transwell结果显示,con-a、con-b和siRNA组穿膜细胞数分别为（38 ±4）、（40±3）和（8±1）个（P＜0.05）.结论 假基因Cr-3在大肠癌细胞增殖侵袭中起着重要的作用；采用RNA干扰下调Cr-3表达可抑制大肠癌细胞增殖和侵袭.
Objective To study the effects of RNA interference pseudogene Cripto-3 （Cr-3） on proliferation and invasion of colon cancer cells.Methods Cr-3 small interfering RNAs （siRNA,S1,S2 and S3） was built and synthesized by using chemical methods.The SW620 cells were classified into three groups：blank control group （con-a）,the empty plasmid group （con-b） and Cr-3 siRNA group （siRNA）.Among them,the siRNA group was transfected with Cr-3 siRNA.The cell viability of each group was measured by methyl thiazol tetrazolium （MTT） assay.The invasion ability was evaluated by transwell method.All the above were for statistical analysis.Results S1,S2 and S3 could downregulate Cr-3 mRNA with the inhibition rate of 83.5%,61.6% and 42.8% respectively.After SW620 cells were transfected by S1,MTT assay revealed that the relative proliferation rate in con-a,con-b and Cr-3 siRNA groups at72 h was （0.927 ±0.037）%,（0.922 ±0.035）% and （0.663 ±0.018）% （A） respectively （P〈0.05）.Transwell showed that the number of cells penetrating the membrane in con-a,con-b and Cr-3 siR-NA group was 38 ± 4,40 ± 3 and 8 ± 1 respectively （P 〈 0.05）.Conclusion Cr-3 gene might play an important role in proliferation and invasion of human colon cancer cells,and down-regulation of Cr-3 by siRNA could inhibit invasion ability and proliferation of human colon cancer cells.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2013年第11期2305-2307,F0003,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目（81250035）
江苏省＂333工程＂科研基金资助项目（BRA2012103）
江苏省创新基金资助项目（CXLX12-0679）
镇江市社会发展基金资助项目（SH2011031）
关键词
大肠肿瘤
侵袭
RNA干扰
Colon carcinoma
Invasion
RNA interference
