miR-195负性调控VEGFA抑制子宫内膜癌干细胞干性  

MiR-195 inhibits endometrial cancer stemness by negatively targeting VEGFA

作　　者：张玉虹[1] 孙喜斌[1] 石克威 马会[1] 于治芳 陈江平[1] ZHANG Yu-hong;SUN Xi-bin;SHI Ke-wei;MA Hui;YU Zhi-fang;CHEN Jiang-ping(Department of Gynaecology,the First Affiliated Hospital of Beifang College,Zhangjiakou HEBEI 075000,China)

机构地区：[1]河北北方学院附属第一医院妇产科,河北张家口075000

出　　处：《中国新药与临床杂志》2020年第12期747-752,共6页Chinese Journal of New Drugs and Clinical Remedies

摘　　要：目的观察miR-195对子宫内膜癌干细胞干性维持和化疗敏感性的影响,及调控血管内皮生长因子A(VEGFA)的相关作用机制。方法选用人源性子宫内膜癌KLE细胞分成空白对照组、空质粒组、VEGFA组、miR-195组和VEGFA+miR-195组,分别转染pc-VEGFA和miR-195 mimic过表达载体。CCK-8法观察细胞增殖和顺铂杀伤率,Transwell法观察细胞侵袭,流式细胞术检测细胞凋亡,平板克隆形成实验观察克隆形成率,RT-PCR法检测细胞VEGFA和miR-195 mRNA表达,Western blot法检测VEGFA蛋白表达,荧光素酶验证miR-195和VEGFA基因突变的关系。结果与空白对照组和空质粒组相比,miR-195组细胞增殖倍数和侵袭数显著下降,顺铂杀伤率显著升高(P <0.05);VEGFA组细胞增殖倍数和侵袭数均升高,顺铂杀伤率显著下降(P <0.05);VEGFA+miR-195组细胞增殖倍数和侵袭数低于VEGFA组,顺铂杀伤率高于VEGFA组(均P <0.05)。空白对照组和空质粒组顺铂的IC50分别为44.40和46.43 mg·L^-1,miR-195组和VEGFA组分别为67.13和62.59mg·L^-1,VEGFA+miR-195组为28.61mg·L^-1。与空白对照组和空质粒组相比,miR-195组肿瘤干细胞克隆形成率降低,miR-195 mRNA表达增加,VEGFA蛋白和mRNA表达降低(均P <0.05);VEGFA+miR-195组肿瘤干细胞克隆形成率及VEGFA蛋白和mRNA表达均低于VEGFA组(P <0.05)。结论 miR-195可抑制子宫内膜癌KLE细胞增殖和侵袭,抑制干细胞干性,对顺铂敏感性增强,其机制与负性调控VEGFA有关。AIM To observe the effect of miR-195 on stemness maintenance and chemotherapy sensitivity of endometrial cancer stem cells, as well as the related mechanism of regulating vascular endothelial growth factor A(VEGFA). METHODS Human-derived endometrial cancer KLE cell lines were divided into blank control group, empty plasmid group, VEGFA group, miR-195 group, and VEGFA + miR-195 group. pc-VEGFA and miR-195 mimic overexpression vectors were transferred into KLE cells simultaneously or separately. Cell proliferation and cisplatin killing rate were observed by CCK-8 method. Cell invasion was observed by transwell method. RT-PCR was used to detect the mRNA expression of VEGFA and miR-195, and western blot was used to detect the protein expression of VEGFA. The association between miR-195 and VEGFA gene mutation was verified by luciferase method. RESULTS Compared with the blank control group and the empty plasmid group, the cell proliferation and invasion numbers decreased and the cisplatin killing rate increased in the miR-195 group(P < 0.05), while the cell proliferation and invasion numbers increased and the cisplatin killing rate decreased in the VEGFA group(P < 0.05). The cell proliferation and invasion numbers of the VEGFA + miR-195 group were lower and cisplatin killing rate was higher than those of the VEGFA group(P < 0.05). The IC50 of cisplatin in the blank control group and the empty plasmid group were 44.40 and 46.63 mg·L^-1, IC50 of miR-195 group and VEGFA group were 67.13 and 62.59 mg·L^-1, and IC50 of VEGFA+miR-195 group was 28.61 mg·L^-1, respectively. Compared with the blank control group and the empty plasmid group, the clone formation rate of tumor stem cells decreased, miR-195 mRNA expression increased, and VEGFA protein and mRNA expression decreased in the miR-195 group(P < 0.05). The clone formation rate and the VEGFA protein and mRNA expression in the miR-195 group were lower than those in the VEGFA group(P < 0.05). CONCLUSION miR-195 can inhibit the proliferation and invasion of endometrial ca

关 键 词：子宫内膜肿瘤 miR-195 肿瘤干细胞 血管内皮生长因子A 化疗敏感性 

分 类 号：R737.33[医药卫生—肿瘤]