miR-3619-5p靶向PFKFB3抑制结肠癌细胞的增殖、迁移和侵袭  

miR-3619-5p inhibits the proliferation,migration and invasion of colon cancer cells by targeting PFKFB3

作　　者：郭银谋[1] 王玉梅[1] 陈贡斌[1] 岳培茹[1] 周威[1] GUO Yin-mou;WANG Yu-mei;CHEN Gong-bin;YUE Pei-ru;ZHOU Wei(Department of Oncology,the First People's Hospital of Shangqiu,Shangqiu476100,China)

机构地区：[1]商丘市第一人民医院肿瘤一科,商丘476100

出　　处：《现代免疫学》2021年第6期487-494,共8页Current Immunology

摘　　要：为研究miR-3619-5p和6磷酸果糖2激酶/果糖-2,6-二磷酸酶3(6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3,PFKFB3)对结肠癌细胞增殖、迁移和侵袭的影响,采用qRT-PCR检测35例结肠癌组织和癌旁组织中miR-3619-5p和PFKFB3 mRNA的水平。在结肠癌细胞HCT116中转染miR-3619-5p模拟物或si-PFKFB3,以过表达miR-3619-5p或抑制PFKFB3。在结肠癌细胞HCT116中转染miR-3619-5p模拟物和PFKFB3过表达载体(pcDNA-PFKFB3),同时过表达miR-3619-5p和PFKFB3。采用CCK-8试验和Transwell试验分别检测结肠癌细胞HCT116增殖、迁移和侵袭能力,Western blotting检测细胞中PFKFB3、细胞周期蛋白D1(Cyclin D1)、p21、基质金属蛋白酶2(matrix metalloprotease 2,MMP-2)和基质金属蛋白酶9(matrix metalloprotease 9,MMP-9)蛋白水平,双荧光素酶报告系统验证miR-3619-5p与PFKFB3的靶向关系。结果显示,与癌旁组织比较,结肠癌组织中miR-3619-5p水平显著降低(P<0.05),PFKFB3 mRNA和蛋白水平显著升高(P<0.05);过表达miR-3619-5p或抑制PFKFB3均可降低Cyclin D1、MMP-2和MMP-9蛋白表达量,提高p21表达量,降低细胞光密度(D)值、迁移和侵袭细胞数;miR-3619-5p靶向负调控PFKFB3的表达;过表达PFKFB3可逆转miR-3619-5p过表达对HCT116细胞增殖、侵袭和迁移的影响。该研究提示,miR-3619-5p可通过靶向PFKFB3抑制HCT116细胞的增殖、迁移和侵袭,miR-3619-5p可能是结肠癌治疗的潜在分子靶点。To investigate the effects of miR-3619-5 p and 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3(PFKFB3) on the proliferation, migration and invasion of colon cancer cells, qRT-PCR was used to detect the expression levels of miR-3619-5 p and PFKFB3 mRNA in 35 cases of colon cancer and adjacent tissues. Colon cancer HCT116 cells were transfected with miR-3619-5 p mimics or si-PFKFB3 to overexpress miR-3619-5 p or inhibit PFKFB3. The miR-3619-5 p mimic and PFKFB3 overexpression vector(pcDNA-PFKFB3) were co-transfected in colon cancer HCT116 cell to overexpress miR-3619-5 p and PFKFB3 at the same time. The proliferation, migration and invasion abilities of HCT116 cells were determined by CCK-8 assay and Transwell assay, respectively. And the levels of PFKFB3, Cyclin D1, p21, matrix metalloprotease 2(MMP-2) and matrix metalloprotease 9(MMP-9) proteins in the cells were measured by Western blotting. The targeting relationship between miR-3619-5 p and PFKFB3 was verified by the dual-luciferase reporter assay system. The results showed that compared with the adjacent tissues group, the level of miR-3619-5 p in the colon cancer tissues group was decreased(P<0.05), and the levels of PFKFB3 mRNA and proteins were significantly increased(P<0.05). Overexpression of miR-3619-5 p or inhibition of PFKFB3 both reduced the expression levels of Cyclin D1, MMP-2 and MMP-9 proteins, increased the level of p21 protein, and reduced the optical density(D) value of HCT116 cells and the number of migrating and invading cells. miR-3619-5 p targeted and negatively regulated the expression of PFKFB3. Overexpression of PFKFB3 reversed the effect of miR-3619-5 p overexpression on proliferation, invasion and migration of HCT116 cells. It shows that miR-3619-5 p inhibits the proliferation, migration and invasion of HCT116 cells by targeting PFKFB3, and could be a potential molecular target for colon cancer treatment.

关 键 词：结肠癌 微小RNA-3619-5p 6磷酸果糖2激酶/果糖-2 6-二磷酸酶3 增殖 迁移 侵袭 

分 类 号：R392.11[医药卫生—免疫学]