野黄芩素通过miRNA-496和基质细胞衍生因子1对子宫颈癌细胞增殖及迁移的影响  

Effects of scutellaria on the proliferation and migration of cervical cancer cells through miRNA-496 and stromal cell-derived factor 1

作　　者：张刘莲 巫梦雪 王茜茜 李盼 赵玲檬 Zhang Liulian;Wu Mengxue;Wang Qianqian;Li Pan;Zhao Lingmeng(Department of Gynecology&Obstetrics,the Central Hospital of Wuhan,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430014,China)

机构地区：[1]华中科技大学同济医学院附属武汉中心医院妇产科,武汉430014

出　　处：《肿瘤研究与临床》2022年第3期161-165,共5页Cancer Research and Clinic

基　　金：湖北省卫生健康委员会联合基金青年人才项目(WJ2019H172)。

摘　　要：目的探讨野黄芩素通过miRNA-496(miR-496)、基质细胞衍生因子1(SDF-1)对子宫颈癌细胞株HCC94细胞增殖及迁移能力的影响。方法取对数生长期的HCC94细胞,分别加入20μmol/L野黄芩素溶液(野黄芩素组)和二甲基亚砜(DMSO)(对照组)。分别采用CCK-8法和Transwell实验检测两组HCC94细胞增殖和迁移能力,实时荧光定量聚合酶链反应(qRT-PCR)检测miR-496和SDF-1 mRNA的相对表达量。采用生物信息学软件TarBase预测miR-496的靶基因,并用双荧光素酶报告基因实验进行验证。采用蛋白质印迹法检测相关蛋白的表达情况。结果与对照组比较,培养第3、4、5天,野黄芩素组HCC94细胞增殖能力均降低(均P<0.05)。野黄芩素组和对照组HCC94细胞的穿胶细胞数分别为(25±5)个和(134±19)个,野黄芩素组细胞迁移能力较对照组降低(t=5.61,P<0.01)。对照组和野黄芩素组miR-496相对表达量分别为1.07±0.12和11.24±2.75,差异有统计学意义(t=3.68,P<0.01)。双荧光素酶报告基因实验结果证实SDF-1是miR-496的下游靶基因。野黄芩素组和对照组HCC94细胞中SDF-1 mRNA相对表达量分别为0.29±0.05和1.01±0.07,差异有统计学意义(t=7.22,P<0.01)。与对照组比较,野黄芩素促进miR-496表达后,SDF-1蛋白、细胞增殖蛋白细胞周期蛋白依赖激酶3(CDK3)、细胞迁移蛋白Slug和Zeb-2表达均降低。结论野黄芩素可通过miR-496-SDF-1轴抑制子宫颈癌HCC94细胞的增殖及迁移。Objective To explore the effects of scutellarein on the cell proliferation and migration of cervical cancer cell line HCC94 through miRNA-496(miR-496)and stromal cell-derived factor 1(SDF-1).Methods HCC94 cells in the logarithmic growth phase were taken as objects,and 20μmol/L scutellarin solution(scutellarin group)and dimethyl sulfoxide(DMSO)(control group)were added respectively.The CCK-8 method and Transwell experiment were used to detect the proliferation and migration ability of the two groups of HCC94 cells.The relative expression levels of miR-496 and SDF-1 mRNA were detected by real-time quantitative polymerase chain reaction(qRT-PCR).The target gene of miR-496 was predicted by the bioinformatics software TarBase and verified by the dual-luciferase reporter gene assay.Western blot was used to detect the expression of related proteins.Results Compared with the control group,the proliferation ability of HCC94 cells in the scutellarin group was decreased on the 3rd,4th and 5th day of culture(all P<0.05).The number of HCC94 cells in the scutellarin group and the control group were 25±5 and 134±19,respectively,and the cell migration ability of the scutellarin group was lower than that of the control group(t=5.61,P<0.01).The relative expressions of miR-496 in the control group and the scutellarin group were 1.07±0.12 and 11.24±2.75,respectively,and the difference was statistically significant(t=3.68,P<0.01).The dual-luciferase reporter gene assay confirmed that SDF-1 was the downstream targeted gene of miR-496.The relative expressions of SDF-1 mRNA in the scutellarin group and the control group were 0.29±0.05 and 1.01±0.07,respectively,and the difference was statistically significant(t=7.22,P<0.01).Compared with the control group,after scutellarin promoted the expression of miR-496,the expressions of SDF-1 protein,the cell proliferation protein cyclin-dependent kinase 3(CDK3)and the cell migration proteins Slug and Zeb-2 were decreased.Conclusions Scutellarin could inhibit the proliferation and migration

关 键 词：宫颈肿瘤 微RNAS 野黄芩素 细胞增殖 细胞迁移 

分 类 号：R285[医药卫生—中药学]