miR-340通过下调CCND1表达增加结直肠癌细胞对5-Fu的耐药

发布时间：2018-09-04 09:00

【摘要】：目的:探讨细胞周期蛋白D1(cyclin D1)的编码基因CCND1 miR-340介导的逆转结直肠癌细胞对5-氟尿嘧啶(5-Fu)耐药的机制。方法:采用瞬时转染技术将结直肠癌细胞HCT116、SW480株分别转染si-CCND1和miR340-mimic。应用MTT法检测转染后的结直肠癌细胞对5-Fu敏感性的变化,应用双荧光素酶试验验证CCND1对miR340参与的影响结直肠癌细胞对5-Fu敏感性的影响。结果:瞬时转染si CCND1和过表达miR-340后,结直肠癌HCT116和SW480细胞的IC50值均显著低于对照组(10,10 vs 20μmol/L和20,20 vs 40μmol/L,均P0.05)。共转染CCND1 3'UTR野生质粒和miR-340 inhibitor的结直肠癌HCT116和SW48细胞荧光素酶的活性显著高于共转染空载体和mimic细胞(P0.01)。结论:CCND1作为不良因子通过抑制miR340的表达进而发挥增加结直肠癌细胞对5-Fu耐药的作用。
[Abstract]:Aim: to investigate the mechanism of reversal of 5-fluorouracil (5-Fu) resistance in colorectal cancer cells mediated by cyclin D1 (cyclin D1) encoding gene CCND1 miR-340. Methods: HCT116,SW480 cells were transfected into si-CCND1 and miR340-mimic. by transient transfection technique. MTT assay was used to detect the sensitivity of transfected colorectal cancer cells to 5-Fu, and double luciferase assay was used to verify the effect of CCND1 on the 5-Fu sensitivity of colorectal cancer cells involved in miR340. Results: after transient transfection of si CCND1 and overexpression of miR-340, the IC50 values of HCT116 and SW480 cells in colorectal cancer were significantly lower than those in control group (10 10 vs 20 渭 mol/L and 20 20 vs 40 渭 mol/L, P0.05). The luciferase activity of HCT116 and SW48 cells cotransfected with CCND1 3'UTR wild plasmid and miR-340 inhibitor was significantly higher than that of empty vector and mimic cells (P0.01). Conclusion as an adverse factor, the expression of miR340 is inhibited and the resistance to 5-Fu is increased in colorectal cancer cells.
【作者单位】：广州中医药大学附属海南省中医院肿瘤科;南方医科大学中西医结合医院肿瘤中心;
【基金】：广东省自然科学基金资助项目(No.2015A030311005)~~
【分类号】：R735.34

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