目的 探讨白花蛇舌草乙醇提取物(EEHD)对转化生长因子-β1(TGF-β1)体外诱导的人肺腺癌细胞H358间质化过程的干预作用。方法 以5 ng·mL-1的转化生长因子-β1作用于H358细胞24 h,成功构建上皮间质转化模型,此后分别加入低、中、高浓度的白花蛇舌草乙醇提取物作用48 h后,通过蛋白质印迹法与定量聚合酶链反应检测各组细胞上皮标志物E-钙黏蛋白(E-cadherin)及间质细胞标志物波蛋白(vimentin)的蛋白和mRNA的表达。结果 5 ng·mL-1的转化生长因子-β1可成功构建H358细胞上皮间质转化模型,在不同浓度白花蛇舌草乙醇提取物干预后,随浓度升高E-钙黏蛋白表达呈上升趋势,波蛋白表达下降。结论 白花蛇舌草乙醇提取物能部分干预甚至逆转转化生长因子-β1诱导的H358细胞上皮间质转化过程。
Abstract
OBJECTIVE To investigate whether the ethanol extract of Oldenlandia diffusa (EEHD) have impact on the TGF-β1-induced epithelial mesenchymal transformation(EMT)of human lung adenocarcinoma cell H358 in vitro.METHODS The cell model to study EMT of H358 cells was established with 5 ng·mL-1 TGF-β1 for 24 h. Then,after being mixed with low, medium and high concentration of EEHD respectively for 48 h, H358 cells were collected for detection of protein and mRNA expression of epithelial marker E-cadherin and mesenchymal cell marker vimentinby Western-Blot and quantitative PCR.RESULTS The concentration of 5 ng·mL-1 TGF-β1 can successfully induce the transformation from epithelialphenotype to mesenchymal phenotype. The concentration of EEHD is positively correlated with increasing expression of E-cadherin,while negatively correlation with the decreasing expression of vimentin.CONCLUSION The EEHD have partially reversal effect of epithelial mesenchymal transformation on lung adenocarcinoma cell induced by TGF-β1.
关键词
白花蛇舌草乙醇提取物 /
上皮间质转化 /
H358细胞
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Key words
Hedyotis diffusa ethanol extract /
epithelial mesenchymal transition(EMT) /
H358 cell
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中图分类号：
R965
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参考文献
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脚注
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基金
浙江省卫生厅科研基金资助项目(2013KYB187);浙江省中医药管理局科研基金资助项目(2013ZQ014)
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