Department of Gastroenterology, The First Affiliated Hospital of Nanchang University, Jiangxi Clinical Research Center for Gastroenterology, Nanchang.
Department of Gastroenterology, The Second Affiliated Hospital of Nanchang University, Jiangxi Clinical Research Center for Gastroenterology, Nanchang.
Eur J Histochem. 2022 May 10;66(2):3333. doi: 10.4081/ejh.2022.3333.
MicroRNAs (miRNAs/miRs) have aroused increasing attention in colorectal cancer (CRC) therapy. This study is designed for a detailed analysis of the roles of miR-16-5p and forkhead box K1 (FOXK1) in cell angiogenesis and proliferation during CRC in addition to their underlying mechanisms. CRC tissues and colon cancer cell lines (SW620 and HCT8) were investigated. qRT-PCR and Western blot were utilized to evaluate miR-16-5p and FOXK1 expression. Following gain- and loss-of-function assays on miR-16-5p or FOXK1, the effects of miR-16-5p and FOXK1 were assessed on cell angiogenesis and proliferation in CRC cells. A dual-luciferase reporter assay was employed to evaluate the binding relationship of miR-16-5p and FOXK1. Western blot was used to determine the effects of miR-16-5p and FOXK1 on key molecules of the PI3K/Akt/mTOR pathway. Highly expressed FOXK1 and lowly expressed miR-16-5p were observed in CRC cells and tissues. miR-16-5p overexpression or FOXK1 knockdown reduced CRC cell proliferation and angiogenesis of human umbilical vein endothelial cells co-cultured with the supernatant of CRC cells, whereas miR-16-5p silencing or FOXK1 upregulation caused opposite trends. Additionally, miR-16-5p negatively modulated FOXK1 expression. The blockade of the PI3K/Akt/mTOR pathway was triggered by miR-16-5p overexpression or FOXK1 silencing. In conclusion, miR-16-5p hampers cell angiogenesis and proliferation during CRC by targeting FOXK1 to block the PI3K/Akt/mTOR pathway.
微小 RNA（miRNAs/miRs）在结直肠癌（CRC）治疗中引起了越来越多的关注。本研究旨在详细分析 miR-16-5p 和叉头框 K1（FOXK1）在 CRC 细胞血管生成和增殖中的作用及其潜在机制。研究了 CRC 组织和结肠癌细胞系（SW620 和 HCT8）。qRT-PCR 和 Western blot 用于评估 miR-16-5p 和 FOXK1 的表达。在对 miR-16-5p 或 FOXK1 进行功能获得和功能丧失实验后，评估了 miR-16-5p 和 FOXK1 对 CRC 细胞血管生成和增殖的影响。双荧光素酶报告基因实验用于评估 miR-16-5p 和 FOXK1 的结合关系。Western blot 用于确定 miR-16-5p 和 FOXK1 对 PI3K/Akt/mTOR 通路关键分子的影响。在 CRC 细胞和组织中观察到高表达的 FOXK1 和低表达的 miR-16-5p。miR-16-5p 过表达或 FOXK1 敲低减少了与 CRC 细胞上清共培养的人脐静脉内皮细胞的 CRC 细胞增殖和血管生成，而 miR-16-5p 沉默或 FOXK1 上调则导致相反的趋势。此外，miR-16-5p 负调控 FOXK1 表达。miR-16-5p 过表达或 FOXK1 沉默触发了 PI3K/Akt/mTOR 通路的阻断。综上所述，miR-16-5p 通过靶向 FOXK1 阻断 PI3K/Akt/mTOR 通路来抑制 CRC 细胞的血管生成和增殖。
