Resveratrol modulates miRNA machinery proteins in different types of colon cancer cells

E. Becer, Servet Madencioğlu, H. Kabadayı, H. Vatansever
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Abstract

Abstract Objectives Resveratrol (RSV) is a stilbenoid compound that shows anticancer activity in many cancer cells. Exosomes might affect carcinogenesis and the development of colorectal cancer by affecting communication between tumor cells in the tumor microenvironment via their cargo content miRNA. The aim of this study is to determine the effect of RSV on the expression of Dicer, Ago2, eIf2α, CD-9, CD-63, and exosomal miRNA levels in COLO320 and COLO741 colon cancer cell lines. Methods The MTT method was used for cell growth and cytotoxicity in both COLO320 and COLO741 cell lines. Dicer, Ago2, eIF2α, CD-9, and CD-63 antibodies were used for the immunocytochemical evaluation. Total miRNA analysis was performed using a miRCURY Exosome Isolation Kit. Results As a result of immunocytochemical staining, increased CD-63 immunoreactivity was observed in RSV-treated COLO320 cells vs. RSV-treated COLO-741 cells. Dicer immunoreactivity increased after the RSV treatment in COLO320 cells. Higher eIF2α immunoreactivity was observed in RSV-treated COLO741 cells compared to both COLO741 control cells and RSV-treated COLO320 cells. Non-significant decreases were observed in miRNA concentration in RSV-treated COLO320 and COLO741 cells compared to control group cells. Conclusions RSV could increase miRNA biogenesis in COLO320 cancer cells and decrease it in COLO741 cancer cells.
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白藜芦醇调节不同类型结肠癌细胞中的 miRNA 机制蛋白
摘要 目的 白藜芦醇(Resveratrol,RSV)是一种二苯乙烯类化合物,对许多癌细胞具有抗癌活性。外泌体可能通过其所含的 miRNA 影响肿瘤微环境中肿瘤细胞之间的交流,从而影响癌变和结直肠癌的发展。本研究旨在确定 RSV 对 COLO320 和 COLO741 大肠癌细胞系中 Dicer、Ago2、eIf2α、CD-9、CD-63 的表达以及外泌体 miRNA 水平的影响。方法 采用 MTT 法检测 COLO320 和 COLO741 细胞系的细胞生长和细胞毒性。采用 Dicer、Ago2、eIF2α、CD-9 和 CD-63 抗体进行免疫细胞化学评估。使用 miRCURY 外泌体分离试剂盒对总 miRNA 进行了分析。结果 免疫细胞化学染色结果显示,RSV 处理的 COLO320 细胞与 RSV 处理的 COLO-741 细胞相比,CD-63 免疫反应性增加。COLO320 细胞经 RSV 处理后,Dicer 免疫活性增加。与 COLO741 对照细胞和 RSV 处理的 COLO320 细胞相比,RSV 处理的 COLO741 细胞中观察到更高的 eIF2α 免疫反应活性。与对照组细胞相比,RSV 处理的 COLO320 和 COLO741 细胞中的 miRNA 浓度下降不明显。结论 RSV 可增加 COLO320 癌细胞的 miRNA 生物发生,减少 COLO741 癌细胞的 miRNA 生物发生。
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