高脂对HepG2肝癌细胞E2F1、细胞侵袭及迁移的影响

Effect of high free fatty on E2F1 expression and invasion and migration of HepG2 cells

目的:探讨高脂对肝癌细胞E2F1、细胞侵袭及迁移的影响。方法:将HepG2肝癌细胞分为对照组和高脂组,分别用不含游离脂肪酸(FFA)培养基和含1 mmol/L FFA(油酸∶棕榈酸=2∶1)培养基培养;采用划痕实验和Transwell迁移、侵袭实验来比较两组细胞的迁移和侵袭能力;采用Western blotting实验和RT-qPCR实验检测E2F1和上皮间质转化(EMT)相关标志物(α-SMA、Vimentin、β-catenin、E-cadherin)的蛋白表达以及mRNA水平;使用Spearman相关分析E2F1与EMT相关标志物的相关关系。结果:与对照组比较,高脂组细胞的划痕愈合率较对照组明显增加、迁移的细胞数量、侵袭的细胞数量均增多(均P<0.05);与对照组比较,高脂组细胞中E2F1、α-SMA、Vimentin、β-catenin的mRNA及其蛋白相对表达量升高,而E-cadherin的mRNA及其蛋白相对表达量降低(均P<0.05);E2F1与α-SMA、Vimentin、β-catenin呈正相关关系(均P<0.05),E2F1与E-cadherin呈负相关关系(P<0.05)。结论:高脂可诱导HepG2肝癌细胞发生EMT并增强细胞的侵袭迁移能力,促进HepG2细胞中E2F1的表达。

Objective:To investigate the effect of high free fat acids(FFA) on E2F1 expression in hepatocellular carcinoma cells,and the potential molecular mechanism of high FFA promoting the invasion and metastasis of hepatocellular carcinoma cells.Methods:HepG2 cells were divided into control group and high FFA group,cultured in free fat acid free medium and 1 mmol/L FFA(oleic acid:palmitic acid = 2:1) medium,respectively.The migration and invasion ability of the two groups of cells were compared by wound healing assay and Transwell assay.The protein and mRNA expressions of E2F1 and EMT-related markers(α-SMA,Vimentin,β-catenin,Ecadherin) were detected by western blotting and RT-qPCR,respectively.Spearman correlation analysis was used to analyze the relationship between E2 F1 and EMT related markers.Results:Compared with the control group,the wound healing rate,the number of migrating cells and invasive cells,and the mRNA and protein expressions of E2F1,α-SMA,Vimentin and β-catenin in high FFA group were significantly increased,while the mRNA and protein expression of E-cadherin were significantly decreased(P<0.05).E2F1 was positively correlated with α-SMA,Vimentin,and β-catenin,and E2F1 was negatively correlated with E-cadherin(P<0.05).Conclusion:High FFA can promote the expression of E2F1 in HepG2 cells,induce EMT and enhance the invasion and migration of HepG2 cells.