沉默Rictor对肝癌细胞体外血管生成、成瘤能力及P4HB/Hedgehog表达的影响

Effect of Silencing Rictor on Angiogenesis,Tumorigenesis and P4HB/Hedgehog Expression of Hepatocellular Carcinoma Cells in Vitro

目的探究沉默Rictor对肝癌细胞体外血管生成、成瘤能力及P4HB/Hedgehog表达的影响。方法取肝癌细胞,将其分为肝癌细胞组(LC组)、肝癌细胞+Rictor-NC组(NC组)、肝癌细胞+siRictor组(SR组),荧光显微镜下观察各组转染情况,实时定量PCR法检测Rictor相关表达,细胞三维培养观察细胞形成拟态血管能力,裸鼠成瘤实验观察成瘤能力,免疫印迹法检测血管生成相关指标及P4HB/Hedgehog表达。结果LC组无转染,而NC组、SR组均可见绿色荧光,说明转染成功,且转染率均达90%以上,说明转染的细胞较为稳定;SR组细胞Rictor mRNA表达、血管形成数量、裸鼠瘤体生长曲线、血管内皮生长因子、P4HB、Hedgehog蛋白表达明显降低(P<0.05),说明沉默Rictor可促进肝癌细胞凋亡并抑制瘤体生成。结论沉默Rictor可显著抑制肝癌细胞体外血管生成,降低裸鼠成瘤能力,并抑制P4HB/Hedgehog表达。

Objective To investigate the effect of silencing Rictor on angiogenesis,tumorigenesis and expression of P4HB/Hedgehog in hepatocellular carcinoma cells in vitro.Methods The hepatocellular carcinoma cells were collected and divided into liver cancer cell group(LC group),liver cancer cell+Rictor-NC group(NC group)and liver cancer cell+siRictor group(SR group).The transfection of each group was observed under fluorescence microscope,and the expression of Rictor was detected by real-time PCR.The ability of the cells to form mimetic vessels was observed by three-dimensional culture,and the tumor-forming ability was observed by tumor-forming assay in nude mice.The angiogenesis related indicators and the expression of P4HB/Hedgehog were detected by western blotting.Results There was no transfection in LC group,but green fluorescence was seen in NC group and SR group,indicating successful transfection;and the transfection rate was above 90%,indicating that the transfected cells were stable.The expression of Rictor mRNA,the number of angiogenesis,tumor growth curve,vascular endothelial growthfactor,P4HB and Hedgehog protein in the SR group were significantly decreased(P<0.05),indicating that Rictor silence can promote apoptosis of hepatocellular carcinoma cells and inhibit tumor formation.Conclusion Silencing Rictor can significantly inhibit the angiogenesis of hepatocellular carcinoma cells in vitro,reduce tumorigenesis in nude mice,and inhibit the expression of P4HB/Hedgehog.