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- 2016
白花蛇舌草总黄酮对TGF-β1诱导的肝癌MHCC97-H细胞EMT的逆转作用及其机制
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Abstract:
摘要：目的 探讨白花蛇舌草总黄酮(FOD)对TGF-β1诱导肝癌细胞MHCC97-H上皮细胞间质转化(EMT)的逆转作用及其相关机制。方法 用TGF-β1诱导MHCC97-H细胞建立EMT模型，再将其分为5组：正常对照组、TGF-β1组、TGF-β1+FOD组、TGF-β1+5-FU组及TGF-β1+FOD+5-FU组，分别干预48？@h，Transwell侵袭小室法检测细胞体外侵袭能力，Western blot法检测各组细胞中E-cadherin、vimentin蛋白的表达。结果 与正常细胞形态相比，TGF-β1诱导后细胞呈现明显的长梭形，且侵袭能力增强(P=0.02)；给予药物处理后，TGF-β1+FOD组、TGF-β1+5-FU组细胞的穿透能力较TGF-β1组减弱(P=0.03、P=0.02)，且联合用药组减弱程度更加明显(P=0.01)；Western blot结果显示，TGF-β1组细胞中E-cadherin蛋白表达显著降低(P=0.01)，vimentin蛋白的表达显著增加(P=0.01)，TGF-β1+FOD组、TGF-β1+5-FU组E-cadherin蛋白表达有所上调(P=0.03、P=0.02)，vimentin蛋白的表达有所下调(P=0.04、P=0.03)，且联合组变化更为显著(P=0.01)。结论 FOD能够逆转TGF-β1诱导的肝癌MHCC97-H细胞的上皮间质转化，其机制可能与其抑制TGF-β1诱导的E-cadherin蛋白下调及上皮细胞间质转化相关。
ABSTRACT: Objective To investigate the effects of total flavones of oldenlandia diffusa (FOD) on epithelial-mesenchymal transition in hepatocellular cancer cell line MHCC97-H. Methods TGF-β1 induced EMT in routinely cultured liver cancer cell line MHCC97-H; then MHCC97-H cell was divided into 5 groups: normal control group, TGF-β1 group, TGF-β1+FOD group, TGF-β1+5-FU group, and TGF-β1+ FOD+5-FU group. After 48？@h of treatment, the invasion ability of MHCC97-H cell was detected by Transwell; the proteins of E-cadherin and vimentin were determined by Western blot. Results Compared with the normal form of MHCC97-H cell line, the cell had obvious long fusiform after TGF-β1 induction, and the invasion ability enhanced (P=0.02). But after treatment, the invasion ability of MHCC97-H cell decreased in FOD group and 5-FU group compared with that in TGF-β1 group (P=0.03, P=0.02), and decreased more significantly in FOD+5-FU group (P=0.01). The expression of E-cadherin at the protein level decreased significantly (P=0.01) in TGF-β1 group, which was abolished in FOD group (P=0.03) and 5-FU group (P=0.02). The expression of vimentin at the protein level increased significantly (P=0.01) in TGF-β1 group, which was abolished in FOD group (P=0.04) and 5-FU group (P=0.03) and more obviously in FOD+5-FU group (P=0.01). Conclusion FOD can reverse the invasion of MHCC97-H cells in EMT induced by TGF-β1 through decreasing the expression of E-cadherin protein and inhibiting the epithelial-mesenchymal transition of MHCC97-H cell
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