薯蓣皂甙元通过miR-145-5p/KLF5/HIF-1α轴抑制结直肠癌细胞增殖、迁移和侵袭  

Diosgenin inhibiting the proliferation,migration and invasion of colorectal cancer cells through miR-145-5p/KLF5/HIF-1αaxis

作　　者：罗育春 罗育连[1] 徐园园 唐清珠[1] Luo Yuchun;Luo Yulian;Xu Yuanyuan;Tang Qingzhu(Department of Anorectology,Chenzhou First People’s Hospital,Chenzhou 423000,China)

机构地区：[1]郴州市第一人民医院肛肠科,423000

出　　处：《中华普通外科学文献（电子版）》2021年第3期178-184,共7页Chinese Archives of General Surgery(Electronic Edition)

摘　　要：目的探究薯蓣皂甙元对结直肠癌细胞的影响及其可能的作用机制。方法体外培养人结直肠癌细胞SW480和人正常结肠上皮细胞CCD-841CoN,对数期生长的SW480细胞分组进行转染和慢病毒感染;RT-PCR检测SW480细胞中miR-145-5p表达、KLF5和HIF-1αmRNA表达;Western blotting检测细胞中KLF5和HIF-1α蛋白表达;CCK-8检测细胞活力;细胞克隆形成实验检测细胞增殖能力;Transwell实验检测细胞迁移和侵袭能力;荧光素酶报告实验检测miR-145-5p对KLF5表达的调控作用。结果与正常组相比,癌细胞组中KLF5和HIF-1α的mRNA和蛋白表达均显著上调(P<0.05),miR-145-5p表达显著下调(P<0.05);与常氧组相比,缺氧组细胞中KLF5、HIF-1α的mRNA和蛋白表达均显著上调(P<0.05);薯蓣皂甙元(10、20、40、80、160μmol/L)可抑制SW480细胞活力(P<0.05);与对照组相比,薯蓣皂甙元组SW480细胞中KLF5和HIF-1α的mRNA和蛋白表达、细胞克隆数、迁移和侵袭细胞数均显著下调(P<0.05),上调KLF5可逆转薯蓣皂甙元对SW480细胞的作用,上调HIF-1α的表达(P<0.05);miR-145-5p靶向调控KLF5;与对照组相比,薯蓣皂甙元组miR-145-5p表达显著上调(P<0.05),KLF5蛋白表达下调(P<0.05);与薯蓣皂甙元组相比,薯蓣皂甙元组+miR-145-5p抑制物组miR-145-5p表达显著下调(P<0.05),KLF5蛋白表达上调(P<0.05)。结论薯蓣皂甙元通过miR-145-5p/KLF5/HIF-1α轴抑制结直肠癌细胞增殖、迁移和侵袭。Objective To explore the effect of diosgenin on colorectal cancer cells and its possible mechanism.Methods Human colorectal cancer cells SW480 and human normal colonic epithelial cells CCD-841CoN were cultured in vitro.RT-PCR was used to detect miR-145-5p expression,KLF5 and HIF-1αmRNA expression.Western blotting was used to detect KLF5 and HIF-1αprotein expression;and CCK-8 was used to detect cell viability.Colony formation test was used to detect cell proliferation.Transwell test was used to detect cell migration and invasion,and luciferase report test to detect the regulatory effect of miR-145-5p on KLF5 expression.Results Compared with the normal group,the mRNA and protein expressions of KLF5 and HIF-1αin cancer cell group were significantly up-regulated(P<0.05),miR-145-5p expression was significantly down-regulated(P<0.05).Compared with the normoxia group,the mRNA and protein expressions of KLF5 and HIF-1αin the hypoxia group were significantly up-regulated(P<0.05).Diosgenin(10,20,40,80,160μmol/L)could inhibit the vitality of SW480 cells(P<0.05).Compared with the control group,the mRNA and protein expression of KLF5 and HIF-1α,the number of cell clones and the number of invasive and migration SW480 cells in the diosgenin group were significantly down-regulated(P<0.05).Up-regulation of KLF5 reversed the effect of diosgenin on SW480 cells and up-regulated the expression of HIF-1α(P<0.05);miR-145-5p targeted regulation of KLF5.Compared with the control group,the expression of miR-145-5p in the diosgenin group was significantly up-regulated(P<0.05),and the expression of KLF5 protein was down-regulated(P<0.05).Compared with the diosgenin group,themiR-145-5p expression in the diosgenin group+miR-145-5p inhibitor group was significantly down-regulated(P<0.05),and KLF5 protein expression was up-regulated(P<0.05).Conclusion Diosgenin inhibits the proliferation,migration and invasion of colorectal cancer cells through the miR-145-5p/KLF5/HIF-1αaxis.

关 键 词：薯蓣皂甙元 结直肠癌 miR-145-5p Krüppel样因子5 缺氧诱导因子1Α 

分 类 号：R735.34[医药卫生—肿瘤]