Department of Gastrointestinal Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, PR China; Department of Anorectal Surgery, Nanchong Central Hospital, Nanchong, Sichuan 637000, PR China.
Department of Pharmacy, Nanchong Central Hospital, Nanchong, Sichuan 637000, PR China.
Biomed Pharmacother. 2019 Jan;109:208-216. doi: 10.1016/j.biopha.2018.10.048. Epub 2018 Nov 2.
The aim of this study was to investigate the role and specific molecular mechanism of miR-31-5 in colorectal cancer. The relative expression of miR-31-5p and NUMB in colorectal cancer tissues was analyzed by qRT-PCR. To knock down the expression of miR-31-5p, the transfection of miR-31-5p inhibitor was performed. The transfection with miR-31-5p mimic was used for miR-31-5p overexpression and pcDNA3.0-NUMB plasmid was used for NUMB overexpression. CCK-8 assay was used to analyze the cell proliferation. Flow cytometry was used to evaluate the cell apoptosis and cell cycle. Matrigel invasion assay was performed to assess the invasion potency and migration assay was performed to assess the migration potency. Hoechst 33258 staining assay was performed to analyze the cell apoptosis of HT29 cells after the indicated transfection. Luciferase activity assays were performed to confirmed the potential binding site for miR-31-5p in 3'-UTR region of NUMB. MiR-31-5p is highly expressed in colorectal cancer and is critical for the cell proliferation, cell cycle, migration, invasion and apoptosis. NUMB is target of miR-31-5p and NUMB overexpression inhibited the cell proliferation, migration, invasion and induced cell cycle arrest and apoptosis in HT29 colorectal cancer cells. In conclusion, miR-31-5p promoted the cell growth, migration and invasion by targeting NUMB in colorectal cancer cells.
本研究旨在探讨 miR-31-5 在结直肠癌中的作用及具体分子机制。采用 qRT-PCR 分析结直肠癌组织中 miR-31-5p 和 NUMB 的相对表达。通过转染 miR-31-5p 抑制剂下调 miR-31-5p 的表达,转染 miR-31-5p 模拟物上调 miR-31-5p 的表达,同时转染 pcDNA3.0-NUMB 质粒过表达 NUMB。CCK-8 法分析细胞增殖,流式细胞术评估细胞凋亡和细胞周期,Matrigel 侵袭实验评估侵袭能力,迁移实验评估迁移能力,Hoechst 33258 染色实验分析转染后 HT29 细胞的凋亡情况,荧光素酶活性实验验证 NUMB 3'-UTR 区 miR-31-5p 的潜在结合位点。结直肠癌中 miR-31-5p 高表达,对细胞增殖、细胞周期、迁移、侵袭和凋亡至关重要。NUMB 是 miR-31-5p 的靶点,NUMB 过表达抑制 HT29 结直肠癌细胞的增殖、迁移和侵袭,并诱导细胞周期停滞和凋亡。综上所述,miR-31-5 通过靶向 NUMB 促进结直肠癌细胞的生长、迁移和侵袭。