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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 effects of siRNA on the proliferation and invasion of human hepatocellular carcinoma cell line SK-HEP-1 by targeting growth hormone receptor (GHR). Methods siRNA targeting GHR expression was synthesized and transfected into SK-HEP-1 cells by Gen MuteTM in vitro. The cells were divided into three groups: blank control group (untransfected siRNA), negative transfected control group Transfection of nonspecific siRNAs) and specific transfection groups (transfection of siRNAs targeting GHR-suppressed expression). Real-time PCR and Western blot were used to detect the expression of GHR m RNA and protein in three groups of cells. The proliferation of three groups of cells was detected by CCK-8 assay. The invasion and migration of three groups of cells were detected by Transwell assay. Results Compared with the negative control group, siRNA transfected by specific siRNA reduced the expression of GHR mRNA and protein in SK-HEP-1 cells. The expression of GHR mRNA and protein in SK-HEP-1 cells was significantly decreased -8 OD value decreased significantly (P <0.05), transwell number of cells was significantly reduced (P <0.05). Conclusion siRNA targeted inhibition of GHR expression, SK-HEP-1 hepatocellular carcinoma cell proliferation and invasion and migration were weakened.