RNA干扰技术沉默GP73对人肝癌HepG2细胞迁移和侵袭的影响
Effects of silencing GP73 with siRNA interference on the migration and invasion of human hepatoma HepG2 cells

目的 ·探讨RNA干扰技术（RNAi）沉默人肝癌HepG2细胞中高尔基体蛋白73(GP73)基因表达后，对细胞迁移和侵袭能力的影响。 方法 ·应用阳离子脂质体法将设计合成的GP73小干扰RNA（siRNA）瞬时转染HepG2细胞。HepG2细胞分为空白对照组、阴性对照组和GP73 siRNA干扰组。各组培养48 h后，采用实时定量 PCR（qRT-PCR）和ELISA法检测转染后HepG2细胞中GP73 mRNA和蛋白表达水平变化，Transwell迁移实验、划痕愈合实验及侵袭实验分别观察迁移和侵袭能力改变，qRT PCR和Western blotting法检测基质金属蛋白酶2（MMP-2）和MMP-9的表达情况。 结果 · GP73 siRNA能显著降低HepG2细胞中GP73 mRNA和蛋白的表达，转染后细胞的迁移和侵袭能力明显下降，其MMP-2和MMP-9 mRNA和蛋白表达水平均明显降低。 结论 · siRNA干扰介导GP73基因沉默可抑制人肝癌HepG2细胞的迁移和侵袭，其机制可能与下调MMP-2和MMP-9的表达水平有关。
： Objective · To explore the effects of silencing Golgi protein 73 (GP73) with RNA interference (RNAi) on the migration and invasion ability of human hepatoma HepG2 cells. Methods · GP73 small interfering RNA (siRNA) synthesized with cathodolyte liposome transfection method was used to transiently transfected into HepG2 cells. HepG2 cells were assigned to the blank control group, the negative control group, and the GP73 siRNA group. Cells were cultured for 48 hours. Real time quantitative PCR (qRT-PCR) and ELISA were used to detect the changes in mRNA and protein expressions of GP73 in HepG2 cells after transfection. Changes in migration and invasion of HepG2 cells were observed using Transwell migration assay, wound healing assay, and Transwell invasion assay. qRT-PCR and Western blotting were used to measure the expressions of matrix metalloproteinase 2 (MMP-2) and MMP-9. Results · GP73 siRNA significantly down-regulated the mRNA and protein expressions of GP73 in HepG2 cells. The migration and invasion of HepG2 cells and mRNA and protein expressions of MP-2 and MMP-9 were significantly decreased after transfection. Conclusion · siRNA interference-mediated GP73 silencing can inhibit the migration and invasion of human hepatoma HepG2 cells. The mechanism may be related to the down-regulation of MMP-2 and MMP-9