Department of Gastroenterology, Affiliated Hospital of Inner Mongolia Medical University, Hohhot, Inner Mongolia, P.R. China.
Eur Rev Med Pharmacol Sci. 2020 Feb;24(4):1794-1806. doi: 10.26355/eurrev_202002_20357.
The application of intestinal microflora is involved in various cancers; however, researches reporting the potential of metabolites of intestinal microflora (MIM) on biological activities of colon cancer (CC) cells are unavailable. This study was designed to testify the functions of MIM on CC cells and its mechanism.
qRT-PCR/Western blot were applied to test the expression levels of miR-192-5p and BMPR2 in human colonic epithelial cells and CC cells (HCT116, SW480). The effects of MIM, mimics-miR-192-5p or inhibitors-miR-192-5p on mRNA and protein expressions of miR-192-5p and BMPR2 were verified by qRT-PCR and Western blot. MTT assay for CC cell viability, flow cytometry for CC cells apoptosis rate, and cell scratch and cell chamber served for the analysis of invasion and migration ability of CC cells. The relationship between miR-192-5p and BMPR2 was validated employing Luciferase reporter gene assay.
Compared with human normal colonic epithelial cells, HCT116 and SW480 cells had lower expression of miR-192-5p and higher expression of BMPR2 (p < 0.01). MIM and mimics-miR-192-5p could enhance cell apoptosis and suppress the migration and proliferation of CC cells. MIM were also found to up-regulate miR-192-5p and down-regulate the expression levels of BMPR2 and p-LIMK2 (p < 0.01). Transfection of inhibitors-miR-192-5p reversed the inhibitory effect of MIM on CC cells.
MIM could up-regulate miR-192-5p to inhibit CC cell growth via down-regulating BMPR2 and inhibiting the activity of RhoA-ROCK-LIMK2 pathway.
肠道微生物群的应用涉及多种癌症；然而，目前尚无关于肠道微生物群代谢物（MIM）对结肠癌（CC）细胞生物学活性的潜在作用的研究报告。本研究旨在验证 MIM 对 CC 细胞的作用及其机制。
采用 qRT-PCR/Western blot 检测人结肠上皮细胞和 CC 细胞（HCT116、SW480）中 miR-192-5p 和 BMPR2 的表达水平。通过 qRT-PCR 和 Western blot 验证 MIM、miR-192-5p 模拟物或 miR-192-5p 抑制剂对 miR-192-5p 和 BMPR2 的 mRNA 和蛋白表达的影响。MTT 法检测 CC 细胞活力，流式细胞术检测 CC 细胞凋亡率，细胞划痕和细胞室分析 CC 细胞侵袭和迁移能力。利用荧光素酶报告基因检测验证 miR-192-5p 和 BMPR2 之间的关系。
与正常人结肠上皮细胞相比，HCT116 和 SW480 细胞中 miR-192-5p 的表达水平较低，BMPR2 的表达水平较高（p<0.01）。MIM 和 miR-192-5p 模拟物可增强 CC 细胞凋亡，抑制 CC 细胞迁移和增殖。还发现 MIM 可上调 miR-192-5p，下调 BMPR2 和 p-LIMK2 的表达水平（p<0.01）。转染 miR-192-5p 抑制剂可逆转 MIM 对 CC 细胞的抑制作用。
MIM 可通过下调 BMPR2 并抑制 RhoA-ROCK-LIMK2 通路活性来上调 miR-192-5p 抑制 CC 细胞生长。
