摘要
目的 观察肝素酶（hparinase,Hpa）小干扰RNA（small-interfering RNA,siRNA）对胰腺癌侵袭力的影响.方法 设计3条Hpa干扰靶序列：Hpa1-siRNA,Hpa2-siRNA,Hpa3-siRNA,稀释退火后分别与线性化pGenesil-1.1质粒表达载体的连接制成pGenesil1.1/Hpa1-siRNA、pGenesil1.1/Hpa2-siRNA、pGenesil1.1/Hpa3-siRNA重组质粒.然后将上述目的基因转染SW1990胰腺癌细胞,并设空白对照和阴性对照组.RT-PCR检测Hpa-mRNA表达,Western blot检测Hpa蛋白表达.Transwell小室检测SW1990细胞体外侵袭力.结果 Hpa-mRNA和Hpa蛋白在基因转染前后的表达,NC和HK组无明显差别,Hpa1-siRNA,Hpa2-siRNA,Hpa3-siRNA三组均明显下降,Hpa2-siRNA抑制效率优于Hpa1-siRNA和Hpa3-siRNA（P＜0.05）.Hpa2-siRNA明显抑制SW1990胰腺癌细胞侵袭力,抑制率均达42.6%.结论 Hpa2-siRNA是Hpa特异性干扰序列,对SW1990胰腺癌细胞侵袭力具有明显抑制作用.
Objective To observed invasion inhibiting effect of Hpa small - interfering RNA （siRNA） on pancreatic cancer ceils. Methods Three designed target genes （ Hpal - siRNA, Hpa2 - siRNA, Hpa3 - siRNA） fragments were diluted, annealed and connected with linear plasmid expression vector pGenesil - 1.1, then pGenesill. 1/Hpal - siRNA, pGenesil1. 1/Hpa2 - siRNA and pGenesill. 1/Hpa3 - siRNA were reconstructed. The Hpa - siRNA was transfected into SW1990 pancreatic cancer cell by DOTAP Liposomes meanwhile the empty control and negative control group were designed. Those target genes were respectively transfected into Hpa - mRNA and Hpa protein expression was detected by using RT - PCR and Western blot respectively. Transwell cab model was employed to test the ability of SW1990 cell. Results Expression of Hpa - mRNA and Hpa - protein in SW1990 were obviously suppressed by Hpal - siRNA, Hpa2 - siRNA, and Hpa3 - siRNA, and efficacy in Hpa2. - siRNA was better than in Hpal - siRNA and Hpa3 - siRNA （ P 〈 0. 05 ）. Invasive ability of pancreatic cancer cells was obviously inhabited by Hpa2 - siRNA （ invasive inhibitory rates reached 42. 6% ）. Conclusions Hpa2 - siRNA is the specific Hpa - siRNA sequence and has obvious effect on pancreatic cancer invasion.
出处
《浙江临床医学》
2011年第5期483-487,共5页
Zhejiang Clinical Medical Journal
基金
浙江省研究生创新科研项目（YK2008071）
