Karimi Dermani Fatemeh, Saidijam Massoud, Amini Razieh, Mahdavinezhad Ali, Heydari Korosh, Najafi Rezvan
Research Center for Molecular Medicine, Hamadan University of Medical Sciences, Hamadan, Iran.
Endometrium and Endometriosis Research Center, Hamadan University of Medical Sciences, Hamadan, Iran.
J Cell Biochem. 2017 Jun;118(6):1547-1555. doi: 10.1002/jcb.25816. Epub 2016 Dec 13.
colorectal cancer (CRC) is one of the most common malignancies, associated with high rates of relapse. A notable challenge in treatment is low response rate to current therapies for advanced CRC. The miR-200c plays an essential role in tumor suppression by inhibiting epithelial-mesenchymal transition (EMT). Resveratrol, a natural compound found in red wine, reveals anti-cancer properties in several types of cancers such as CRC. The aim of current study was to evaluate the effects of resveratrol on proliferation, apoptosis, and invasion of HCT-116 cells and also expression of EMT-related genes in presences or absence of miR-200c.
the effect of resveratrol on viability was examined by MTT assay. LNA-anti-miR-200c transfection of HCT-116 cells was carried out in a time dependent manner. Then, the expression of miR-200c and EMT-related genes were quantified by qRT-PCR. Further, expression of EMT-related proteins, apoptosis, and invasion were analyzed by Western blot, Annexin V/PI staining and scratch test, respectively.
resveratrol could significantly inhibit viability of HCT-116 cells. LNA-anti-miR-200c suppressed the endogenous miR-200c in transfected cells compared with the control. qRT-PCR and Western blot analysis of LNA-anti-miR-200c transfected cells revealed a considerable increase in vimentin and ZEB-1 expression, with a concomitant reduction in E-cadherin expression level. Migration of HCT-116 cells increased, and apoptosis significantly reduced in transfected cells. While, resveratrol could entirely reverse these changes by modulation of miR-200c expression.
our findings revealed a major role of resveratrol in apoptosis, invasion, and switching of EMT to MET phenotype through upregulation of miR-200c in CRC. J. Cell. Biochem. 118: 1547-1555, 2017. © 2016 Wiley Periodicals, Inc.
结直肠癌(CRC)是最常见的恶性肿瘤之一,复发率高。治疗中的一个显著挑战是晚期CRC对当前疗法的低反应率。miR-200c通过抑制上皮-间质转化(EMT)在肿瘤抑制中起重要作用。白藜芦醇是红酒中发现的一种天然化合物,在几种癌症如CRC中显示出抗癌特性。本研究的目的是评估白藜芦醇在有或无miR-200c存在的情况下对HCT-116细胞增殖、凋亡和侵袭的影响以及EMT相关基因的表达。
通过MTT法检测白藜芦醇对细胞活力的影响。以时间依赖性方式对HCT-116细胞进行锁核酸抗miR-200c转染。然后,通过qRT-PCR定量miR-200c和EMT相关基因的表达。此外,分别通过蛋白质印迹法、膜联蛋白V/碘化丙啶染色和划痕试验分析EMT相关蛋白的表达、凋亡和侵袭情况。
白藜芦醇可显著抑制HCT-116细胞的活力。与对照组相比,锁核酸抗miR-200c抑制了转染细胞中的内源性miR-200c。对锁核酸抗miR-200c转染细胞的qRT-PCR和蛋白质印迹分析显示波形蛋白和锌指蛋白E盒结合因子1(ZEB-1)表达显著增加,同时E-钙黏蛋白表达水平降低。HCT-116细胞的迁移增加,转染细胞中的凋亡显著减少。而白藜芦醇可通过调节miR-200c表达完全逆转这些变化。
我们的研究结果揭示了白藜芦醇在CRC中通过上调miR-200c在凋亡、侵袭以及将EMT转换为间充质-上皮转化(MET)表型中起主要作用。《细胞生物化学杂志》118: 1547 - 1555, 2017。© 2016威利期刊公司。