siRNA靶向抑制生长激素受体对肝癌细胞株SK-HEP-1增殖及侵袭能力的影响

Effect of small interfering RNA targeting inhibition of growth hormone receptor with exon 3 deleted on proliferation and invasion of human liver cancer line SK-HEP-1

目的观察siRNA技术靶向抑制生长激素受体(GHR)对人肝癌细胞SK-HEP-1增殖及侵袭能力的影响。方法合成靶向抑制GHR表达的siRNA,通过Gen MuteTM体外转染至人肝癌细胞SK-HEP-1中,将细胞分为三组:空白对照组(未转染siRNA)、阴性转染对照组(转染非特异性的siRNA)和特异性转染组(转染靶向抑制GHR表达的siRNA)。分别利用Real-time PCR方法和Western blot技术检测GHR m RNA和蛋白在三组细胞中的表达,采用CCK-8法检测三组细胞的增殖能力,Transwell法检测三组细胞的侵袭迁移能力。结果与阴性转染对照组相比,特异性转染组siRNA下调了肝癌细胞株SK-HEP-1中GHR m RNA及蛋白的表达;特异性转染组肝癌细胞SK-HEP-1中的CCK-8的OD值明显降低(P<0.05),Transwell穿透的细胞数明显减少(P<0.05)。结论 siRNA靶向抑制GHR表达后,肝癌细胞SK-HEP-1的增殖及侵袭迁移能力减弱。

Objective To investigate the effect of small interfering RNA （siRNA） targeting inhibition of growth hormone receptor with exon 3 deleted （GHR） on proliferation and invasion of human liver cancer cell line SK-HEP- 1. Methods SK- HEP-1 ceils were transfected with siRNA targeting human GHR by GenMuteTM transfection regent. The cells were divided into three groups： control group （non-transfected siRNA） , negative control group （transfected with non-specific siRNA） and specificity group （transfected with expression specifically interfere with GHR siRNA）. Real-time PCR was used to detect the relative expression of GHR mRNA. Western blotting was used to detect the expression level of GHR protein. The cell proliferation was determined by CCK- 8 assay. And the ability of invasion was examined by Transwell assay. Results Compared to the control group, the expression of GHR mRNA and GHR protein could be suppressed obviously by specific siRNA targeting GHR gene, The OD value of CCK-8 of SK-HEP-1 cells was reduced obviously （P〈0.05）. The number of migrating cells was obviously reduced in Transwell test. Conclusion siRNA targeting human GHR could significantly reduce the capability of proliferation migration and invasion of SK-HEP- 1 cells.