抑制UHRF1基因表达对肝癌细胞MHCC-97H迁移的影响及相关机制

Effect of UHRF1 gene inhibition on the migration of hepatoma MHCC-97H cell and its relevant mechanism

目的探讨通过RNA干扰抑制泛素样含PHD和环指域1(UHRF1)基因表达对肝癌MHCC-97H细胞迁移的影响及可能机制。方法合成UHRF1基因特异性小发夹RNA(shRNA),构建稳定干扰UHRF1基因表达的重组慢病毒LV-sh UHRF1(以空载体慢病毒LV-sh NC作为阴性对照),并将其感染MHCC-97H细胞。实时荧光定量PCR法检测UHRF1 mRNA在MHCC-97H细胞中表达情况,蛋白质印迹法检测UHRF1、基质金属蛋白酶-9(MMP-9)和血管内皮生长因子(VEGF)蛋白表达情况,Transwell法检测MHCC-97H细胞迁移能力。结果 LV-sh UHRF1组UHRF1mRNA和蛋白表达量及MMP-9、VEGF蛋白表达量均明显低于LV-sh NC组(P均<0.05),迁移细胞数明显少于LV-sh NC组(P<0.05)。结论通过慢病毒介导的RNA干扰技术抑制UHRF1的表达后可抑制MHCC-97H细胞的迁移能力,机制可能与影响MMP-9和VEGF的表达有关。

Objective It is to investigate the effect of RNA interference targeting the ubiquitin-like with PHD and ringfinger domains 1 （ UHRF1 ） gene on the migration of human hepatoma carcinoma MHCC-97H cells and its relevant mechanism. Methods Synthesize small hairpin RNA （shRNA） targeting UHRF1 to construct recombinant lentiviral vector LV-shUHRF1 which stable knockdown UHRF1, and the recombinant vector was infected into MHCC-97H cells （empty lentivirus LV-shNC as a negative control）. The expression level of UHRF1 mRNA was detected by real-time fluorescence quantitative-PCR, and the expression levels of UHRF1, MMP9 and VEGF were detected by Western blotting. Then the migration of MHCC-97H cells after infection was detected by Transwell assay. Results Compared with LV-shNC group, the UHRFI mRNA and protein expression levels and the of MMP9 and VEGF protein expression levels in MHCC-97H cells after infection with lentivirus LV- shUHRF1 were downregulated （ all P 〈 0.05 ） , the migration of MHCC-97H cells in LV-shUHRF1 group were inhibited significantly （P 〈 0. 05 ）. Conclusion UHRF1 gene silencing by lentivirus-mediated shRNA interference can inhibit MHCC-97H cell migration. This effect may be related to downregulation of MMP9 and VEGF expression.