RNA 干扰畸胎瘤细胞源性生长因子基因对人肝细胞癌 HepG2增殖和侵袭的影响

摘要

目的：探讨 RNA 干扰畸胎瘤细胞源性生长因子（PC cell-derived growth factor，PCDGF）基因对肝细胞癌细胞 HepG2增殖和侵袭的影响。方法采用脂质体法将 PCDGF-shRNA 表达载体瞬时转染人肝细胞癌细胞HepG2，逆转录-聚合酶链反应（reverse transcription-polymerase chain reaction，RT-PCR）和免疫蛋白印迹（Western Blot）检测转染后细胞 PCDGF mRNA 和蛋白的表达，噻唑蓝（thiazolyl blue tetrazolium bromide，MTT）比色法检测转染后 HepG2细胞体外增殖能力，体外侵袭实验检测转染后 HepG2细胞侵袭能力。结果转染后，与未转染组（未转染质粒和脂质体）、脂质体组（转染空脂质体）和阴性质粒组（转染阴性质粒）相比，转染组（转染PCDGF-shRNA 质粒）HepG2细胞的 PCDGF mRNA 和蛋白表达水平明显降低（P ＜0．01），细胞的增殖能力受到抑制（P ＜0．01），细胞的侵袭能力也明显降低（P ＜0．01）。结论 RNA 干扰 PCDGF 基因可抑制肝细胞癌细胞的体外增殖及侵袭能力，PCDGF 基因可能成为肿瘤基因治疗的新靶点。%Objective To investigate the effect of PC cell-derived growth factor(PCDGF) RNA on hepatocellular carcinoma cells HepG2 in vitro .Methods The PCDGF-shRNA expression vector was transfected into hepatocellular carcinoma cells HepG2 by lipofection technology.The expression of PCDGF mRNA and protein were detected by reverse transcription-polymerase chain reaction(RT-PCR)and Western Blot,respectively.The ability of tumor cell proliferation was determined by MTT assay.The effect of PCDGF expression on invasion was also identified by transwell chamber system.Results The expression of PCDGF mRNA and protein were both decreased in the transfection group than in the non-transfection group,liposome group and negative vector group(P <0.01).The MTT showed cell proliferation activity was inhibited in the transfection group (P <0.01).Transwell experiments showed that cell invasion ability was lower in the transfection group compared with that in the non-transfection,liposome and negative vector groups(P <0.01).Conclusion PCDGF RNA interference can inhibit the proliferation and invasion abilities of hepatocellular carcinoma cells in vitro .PCDGF gene may be the new target of gene therapy.