1 Department of Gastrointestinal Surgery, The First Affiliated Hospital of Chongqing Medical University , Chongqing, People's Republic of China .
2 Department of Gastrointestinal Surgery, Nanchong Central Hospital , Nanchong, Sichuan Province, People's Republic of China .
DNA Cell Biol. 2018 May;37(5):501-511. doi: 10.1089/dna.2017.4080. Epub 2018 Mar 9.
microRNA (miR)-92b is an oncogenic miRNA. F-box and WD-40 domain protein 7 (FBXW7/hCdc4) is a tumor suppressor and a target of miR-92b-3p. This study was designed to investigate the effect of miR-92b-3p on colorectal carcinoma (CRC) invasion. The expression levels of miR-92b-3p in human HT29, HCT116, and human fetal colon (FHC) normal cells were detected. HT29 and HCT116 cells were transfected with either miR-92b-3p inhibitor or FBXW7 expression plasmids (pcDNA-FBXW7) and combination of miR-92b-3p and siRNA-FBXW7. Cell viability, migration, invasion, colony formation, cell cycle, and apoptosis in transfected cells were detected using corresponding methods. Moreover, the target relationship between miR-92b-3p and FBXW7 was verified using dual-luciferase reporter assay. miR-92b-3p was upregulated in CRC cells in comparison with FHC cells. Then, we transfected HT29 and HCT116 cells with miR-92b-3p inhibitor or pcDNA-FBXW7 and found decreased cell proliferation, migration, invasion, and colony formation ability, as well as the number of upregulated cells at G1 phase of cell cycle and cell apoptosis. With the cotransfection of miR-92b-3p inhibitor and siRNA-FBXW7, we determined that siRNA-FBXW7 partially attenuated the effect of miR-92b-3p inhibitor on cell behaviors. SiRNA-FBXW7 administration to miR-92b-3p inhibitor-treated cells rebooted cell proliferation, cell migration, invasion, and colony formation. miR-92b-3p inhibition prevented CRC proliferation, invasion, and migration by upregulating FBXW7, which might suggest the potential role of miR-92b-3p in colorectal carcinogenesis and metastasis.
微小 RNA(miR)-92b 是一种致癌 miRNA。F 框和 WD-40 结构域蛋白 7(FBXW7/hCdc4)是一种肿瘤抑制因子,也是 miR-92b-3p 的靶标。本研究旨在探讨 miR-92b-3p 对结直肠癌(CRC)侵袭的影响。检测人 HT29、HCT116 和人胎结肠(FHC)正常细胞中 miR-92b-3p 的表达水平。用 miR-92b-3p 抑制剂或 FBXW7 表达质粒(pcDNA-FBXW7)以及 miR-92b-3p 和 siRNA-FBXW7 的组合转染 HT29 和 HCT116 细胞。用相应的方法检测转染细胞的细胞活力、迁移、侵袭、集落形成、细胞周期和细胞凋亡。此外,利用双荧光素酶报告基因检测验证 miR-92b-3p 和 FBXW7 之间的靶标关系。与 FHC 细胞相比,CRC 细胞中 miR-92b-3p 上调。然后,我们用 miR-92b-3p 抑制剂或 pcDNA-FBXW7 转染 HT29 和 HCT116 细胞,发现细胞增殖、迁移、侵袭和集落形成能力下降,细胞周期 G1 期细胞数量增加,细胞凋亡增加。共转染 miR-92b-3p 抑制剂和 siRNA-FBXW7 后,我们确定 siRNA-FBXW7 部分减弱了 miR-92b-3p 抑制剂对细胞行为的影响。用 miR-92b-3p 抑制剂处理的细胞转染 siRNA-FBXW7 后,细胞增殖、迁移、侵袭和集落形成能力得到恢复。miR-92b-3p 通过上调 FBXW7 抑制 CRC 增殖、侵袭和迁移,这可能表明 miR-92b-3p 在结直肠癌发生和转移中具有潜在作用。