miR-891a-5p对结肠癌细胞肿瘤生物学行为和血管生成潜能的作用

Role of miR-891a-5p on Tumor Biological Behavior and Angiogenic Potential of Colon Cancer Cells

目的:探索miR-891a-5p对结肠癌细胞的增殖、侵袭、迁移、凋亡及血管生成的影响及其作用机制。方法:通过PCR技术筛选miR-891a-5p在FHC,SW480,HCT116,HCT8,和SW620各细胞系中表达最高的细胞SW480,将miR-891a-5p模拟物(miR-891a-5p mimic),miR-891a-5p抑制剂(miR-891a-5p inhibitor)及其相应对照转染至SW480细胞;将SW480分为OE-NC组和OE-NLRP7,CCK-8法测定SW480的增殖能力,Transwell试验测定SW480的侵袭能力,划痕试验测定SW480的迁移能力;Western blot分析VEGF和MMP-9的蛋白表达情况;HUVEC血管形成实验检查血管生成能力。荧光素酶报告基因检测验证miR-891a-5p及NLRP7间的靶向关系;RNA免疫沉淀分析miR-891a-5p及NLRP7之间的作用关系。结果:miR-891a-5p在结肠癌细胞中表达显著高于人正常结肠上皮细胞,NLRP7在结肠癌细胞的表达明显低于正常干细胞,抑制miR-891a-5p使得SW480细胞的增殖、侵袭、迁移及血管生成能力明显下降(P<0.01),VEGF和MMP-9蛋白表达水平显著降低(P<0.01),靶向关系结果显示,miR-891a-5p和NLRP7之间存在靶向作用关系;NLRP7过表达可以降低SW480细胞的增殖、迁移及血管生成能力(P<0.01)。结论:miR-891a-5p可以调控NLRP7促进结肠癌细胞的增殖、侵袭、迁移及血管生成能力加速结肠癌的发展。

Objective:To explore the effects of miR-891a-5p on the proliferation,invasion,migration,apoptosis and angiogenesis of colon cancer cells and its mechanism.Methods:The cell with the highest expression of miR-891a-5p in each cell line of FHC,SW480,HCT116,HCT8,and SW620 was screened by PCR for SW480,and miR-891a-5p mimic(miR-891a-5p mimic),miR-891a-5p inhibitor(miR-891a-5p inhibitor)and their corresponding controls were transfected into SW480 cells.SW480 were divided into OE-NC group and OE-NLRP7,and the proliferative ability of SW480 was determined by CCK-8 method,the invasive ability of SW480 was determined by Transwell test,and the migratory ability of SW480 was determined by scratch test;the protein expression of VEGF and MMP-9 was analyzed by Western blot;and the angiogenic ability was examined by HUVEC angiogenesis assay.The targeting relationship between miR-891a-5p and NLRP7 was validated with luciferase reporter gene assay;the interaction between miR-891a-5p and NLRP7 was analyzed with RNA immunoprecipitation.Results:The expression of miR-891a-5p in colon cancer cells was significantly higher than that in human normal colon epithelial cells,and the expression of NLRP7 in colon cancer cells was significantly lower than that in normal stem cells.Inhibition of miR-891a-5p significantly decreased the proliferation,invasion,migration and angiogenesis of SW480 cells(P<0.01).The protein expression levels of VEGF and MMP-9 were significantly decreased(P<0.01).The results of targeting relationship showed that there was a targeting relationship between miR-891a-5p and NLRP7.NLRP7 overexpression could reduce the proliferation,migration,and angiogenesis of SW480 cells(P<0.01).Conclusion:miR-891a-5p can regulate NLRP7 to promote the proliferation,invasion,migration and angiogenesis of colon cancer cells to accelerate the development of colon cancer.