[关键词]
[摘要]
目的 用分子对接技术探讨苍耳亭在上皮间质转化(EMT)过程中的作用靶点,并通过Western Blotting实验检测其对肝癌HepG2细胞相应靶点蛋白表达的影响。方法 选取在EMT过程中的关键因子蓬乱蛋白(Dhs)、波形蛋白(Vimentin)、Snail、血管内皮生长因子受体3(VEGFR3)为作用靶点,采用分子对接虚拟技术评价苍耳亭与其空间结合能力,并与相应内源性物质烟酰胺腺嘌呤二核苷酸、醋酸离子、黄素腺嘌呤二核苷酸、N-乙酰葡糖胺对比;培养HepG2细胞,给予1、5、20 μmol/L浓度的苍耳亭,利用Western Blotting实验检测其对Dhs、Vimentin、Snail、VEGFR3蛋白表达的影响。结果 分子对接结果显示,苍耳亭与EMT过程中作用靶点均具有一定亲和力,其中与Dhs、Snail、VEGFR3的亲和力高于内源性物质,与Vimentin的亲和能力不及内源性物质;Western Blotting实验结果显示,苍耳亭显著下调Vimentin、Snail、VEGFR3蛋白的表达,显著上调E-cadherin蛋白表达(P<0.05、0.01、0.001)。结论 苍耳亭对肝癌侵袭转移关键因子E-cadherin、Vimentin、Snail、VEGFR3有明显影响,可能是其潜在靶点;分子虚拟对接和Western blotting实验结果具有一定的相似性,能预先提示潜在靶因子。
[Key word]
[Abstract]
objective To explore the role of Xanthatin in the targets of epithelial-mesenchymal transition (EMT) process using molecular docking method, and the effect on the target protein expression of HepG2 cells was detected by Western assay. Method Dhs, Vimentin, Snail and VEGFR3 are critical targets in EMT process, the spatial binding ability of Xanthium was evaluated by molecular docking method, compared with the corresponding endogenous substances:nicotinamide adenine dinucleotide, Acetate ion, flavin adenine dinucleotide, and N-Acetyl-D-glucosamine. HepG2 cells were cultured, and the effects of Xanthatin of 1, 5 and 20 mol/L concentrations on Dhs, Vimentin, Snail and VEGFR3 protein expression were detected by Western Blotting assay. Rusult Molecular docking show that Xanthatin has obvious affinity to key factors of EMT process such as Dhs, Vimentin, and VEGF-R3, higher than that of endogenous substance; and the affinity with Vimentin was less than that of endogenous substance; Western Blotting experiments proved the virtual results. The expression of Vimentin, Snail, VEGFR3 protein was significantly lowered, and the expression of e-cadherin was significantly raised. Conclusion The influence of Xanthatin to key factor e-cadherin, Vimentin, Snail, VEGFR3 are obvious, Which is likely to be a potential target. The results of computer virtual experiment and Western Blotting have certain similarity. Molecular virtual docking can pre hint the potential target factor.
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[基金项目]
江苏省南通市青年基金资助项目(WQ2016056)