Combination of Evodiamine with Berberine Reveals a Regulatory Effect on the Phenotypic Transition of Colon Epithelial Cells Induced by CCD-18Co
Combination of Evodiamine with Berberine Reveals a Regulatory Effect on the Phenotypic Transition of Colon Epithelial Cells Induced by CCD-18Co
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吴茱萸碱与小檗碱的组合揭示了对 CCD-18Co 诱导的结肠上皮细胞表型转变的调节作用
DOI：
10.24920/003670
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
沈维增
中科院分区：
文献类型：
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作者：
黄超;向科明;梁炳君;黄伟旋;张樊君;邵玉婉;王秀莲;刘浩生;沈维增
目的基质细胞之间或基质细胞与癌细胞之间存在转分化。吴茱萸碱和小檗碱是中药复方左金丸的主要药理成分，在重塑肿瘤微环境方面发挥着重要作用。本研究旨在探讨吴茱萸碱和小檗碱联合应用（cBerEvo）对肿瘤相关成纤维细胞诱导的结肠上皮细胞表型转化的影响及其机制方法用人结肠肌成纤维细胞系CCD-18 Co的条件培养液处理人正常结肠上皮细胞系HCoEpiC细胞，诱导上皮细胞向间质细胞转化。相差显微镜观察形态学变化。免疫荧光显微镜下观察上皮-间质转化标志物E-cadherin、vimentin和α-平滑肌肌动蛋白（α-SMA）的表达。通过伤口愈合测定评估迁移。Western blotting检测E-cadherin、vimentin、α-SMA、Snail、ZEB 1和Smads的表达。结果与对照组相比，肿瘤相关成纤维样CCD-18 Co细胞诱导E-cadherin表达下调，vimentin、α-SMA、Snail和ZEB 1表达上调（P<0.05），并促进HCoEpiCs的迁移（P<0.05），Smad 2、p-Smad 2、Smad 3、p-Smad 3和Smad 4在乳腺癌中均呈高表达转化生长因子-β（TGF-β）受体抑制剂LY 364947和cBerEvo以浓度依赖性方式消除这种作用（P<0.05）。结论cBerEvo可通过抑制TGF-β/Smads信号通路的活性来调节CCD-18 Co诱导的结肠上皮细胞分化。
Objective Transdifferentiation exists between stromal cells or between stromal cells and cancer cells. Evodiamine and berberine are predominant pharmacological components of Zuojin pill, a prescription of Traditional Chinese Medicine, playing crucial functions in remolding of tumor microenvironment. This study aimed to explore the effect of combination of evodiamine with berberine (cBerEvo) on the phenotypic transition of colon epithelial cells induced by tumor-associated fibroblasts, as well as the involved mechanisms.Methods Human normal colon epithelial cell line HCoEpiC cells were treated with the prepared conditioned medium of CCD-18Co, a human colon myofibroblast line, to induce epithelial-mesenchymal transition. Phase contrast microscope was used to observe the morphological changes. Epithelial-mesenchymal transition markers including E-cadherin, vimentin and alpha-smooth muscle actin (α-SMA) were observed with immunofluorescence microscopy. Migration was assessed by wound healing assay. Western blotting was used to detect the expressions of E-cadherin, vimentin, α-SMA, Snail, ZEB1 and Smads. Results In contrast to the control, the tumor-associated fibroblasts-like CCD-18Co cells induced down-regulation of E-cadherin and up-regulation of vimentin, α-SMA, Snail and ZEB1 (P<0.05), and promoted migration of HCoEpiCs (P<0.05), with over expression of Smads including Smad2, p-Smad2, Smad3, p-Smad3 and Smad4 (P<0.05), which were abolished by a transforming growth factor-β (TGF-β) receptor inhibitor LY364947 and by cBerEvo in a concentration dependent manner. In addition, cBerEvo-inhibited ratios of p-Smad2/Smad2 and p-Smad3/Smad3 were also dose dependent.Conclusion The above results suggest that cBerEvo can regulate the differentiation of colon epithelial cells induced by CCD-18Co through suppressing activity of TGF-β/Smads signaling pathway.