沉默GPC3对肝癌Huh-7细胞增生、迁移和侵袭能力的影响

Effect of Hippo pathway targeting GPC3 on the proliferation, migration, and invasion abilities of hepatoma Huh-7 cell

目的探讨Hippo通路靶向GPC3后对肝癌Huh-7细胞增生、迁移、侵袭能力的影响。方法采用Western blot检测法及RT-PCR分别测量肝癌Huh-7不同对数生长期的肝癌细胞株GPC3蛋白表达水平及GPC3 mRNA水平,并筛选出可有效沉默GPC3基因的siRNA。将肝癌Huh-7细胞株分为实验组、未转染组及对照组,实验组导入GPC3-siRNA-1633转染Huh-7,其余两组不作处理。EdU实验检查三组细胞增殖率,划痕实验测定各组细胞迁移率,Transwell实验测定各组细胞侵袭能力。结果实验组GPC3 mRNA表达量及GPC3蛋白水平显著低于未转染组及对照组,差异有统计学意义(P<0.05)。实验组Hippo通路中YAP mRNA表达量显著低于未转染组及对照组,差异有统计学意义(P<0.05)。转染48 h后,实验组细胞增生率、迁移率及侵袭率显著低于未转染组及对照组,差异有统计学意义(P<0.05)。结论 GPC3可促进肝癌Huh-7细胞增殖、侵袭及转移,从而促进肝癌细胞生长。CPG3可通过上调Hippo通路中YAP表达水平从而促进肝癌细胞增生。通过沉默GPC3 mRNA后可有效抑制肝癌Huh-7细胞生长。

Objective To investigate effect of Hippo pathway targeting GPC3 on the proliferation, migration, and invasion abilities of hepatoma Huh-7 cell. Methods Western blot assay and RT-PCR were used to measure liver cancer Huh-7 GPC3 protein expression levels in different HCC cell lines in logarithmic growth phase and GPC3 mRNA levels, and can effectively filter out GPC3 gene silencing siRNA. The hepatoma Huh-7 cell lines divided into two groups, non-transfected group and the control group, the experiment group import GPC3-siRNA-1633-transfected Huh-7, do not deal with the other two groups. EdU experimental examination of cell proliferation rate of the three groups, each group was determined experimentally scratch mobility, Transwell experimental determination of each group cell invasion. Results The GPC3 mRNA GPC3 expression and protein levels were significantly lower than non-transfected group and the control group, the difference was statistically significant （P〈0.05）. The expression level of YAP mRNA in Hippo pathway in observation group was significantly lower than the non-transfected group and the control group, the difference was statistically significant （P〈0.05）. After transfection 48 h, cell proliferation rate in the experiment group, mobility and invasion were significantly lower than the non-transfected group and the control group, the difference was statistically significant （P〈0.05）. Conclusion GPC3 can promote liver cancer Huh-7 cell proliferation, invasion and metastasis, thus promoting the growth of liver cancer cells. CPG3 can up-regulate the expression level of YAP in Hippo pathway to promote liver cancer cell proliferation. Through the silence GPC3 mRNA can effectivelynbsp;inhibit the growth of hepatoma Huh-7 cells.