MiR-31-5p promotes the cell growth, migration and invasion of colorectal cancer cells by targeting NUMB
MiR-31-5p promotes the cell growth, migration and invasion of colorectal cancer cells by targeting NUMB
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DOI：
10.1016/j.biopha.2018.10.048
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发表时间：
2019-01-01
影响因子：
7.5
通讯作者：
Wang, Ziwei
中科院分区：
文献类型：
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作者：
Peng, Hong;Wang, Longfei;Wang, Ziwei
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。通过qRT-PCR分析结直肠癌组织中miR-31-5p和NUMB的相对表达量。为了敲低miR-31-5p的表达，进行miR-31-5p抑制剂的转染。用miR-31-5p模拟物转染用于miR-31-5p过表达，pcDNA3.0-NUMB质粒用于NUMB过表达。 CCK-8法用于分析细胞增殖。流式细胞术用于评估细胞凋亡和细胞周期。进行基质胶侵袭测定以评估侵袭效力，并进行迁移测定以评估迁移效力。进行Hoechst 33258染色测定来分析指定转染后HT29细胞的细胞凋亡。进行荧光素酶活性测定以确认 NUMB 3'-UTR 区域中 miR-31-5p 的潜在结合位点。 MiR-31-5p在结直肠癌中高表达，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 的靶标，NUMB 过表达抑制 HT29 结直肠癌细胞的细胞增殖、迁移、侵袭并诱导细胞周期停滞和凋亡。总之，miR-31-5p通过靶向结直肠癌细胞中的NUMB促进细胞生长、迁移和侵袭。
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.