摘要
目的:探讨miR-206对雌激素诱导的ER-α36阳性胃癌(gastric cancer,GC)细胞BGC-823增殖和侵袭的影响及其相关机制。方法:用不同浓度(1、10和100 pmol/L)的雌二醇(estradiol,E_(2))刺激ER-α36阳性BGC-823细胞后,用qPCR法检测miR-206表达水平,MTT法和Transwell实验分别检测细胞的增殖和侵袭能力,WB法检测细胞中CDK14的表达。将miR-206 mimic、miR-NC、pcDNA-CDK14、pcDNA-vector等转染ER-α36阳性BGC-823细胞,并给予100 pmol/L的E_(2)处理后,用MTT法和Transwell小室法分别检测细胞的增殖和侵袭能力,WB法检测细胞中CDK14的表达。用双荧光素酶报告基因实验验证miR-206与CDK14之间的靶向关系。结果:E_(2)能显著降低ER-α36阳性BGC-823细胞中miR-206表达水平(P<0.05或P<0.01)、增强细胞的增殖和侵袭能力(P<0.05或P<0.01)、上调细胞中CDK14的表达水平(P<0.01)。过表达miR-206能显著降低E_(2)诱导的ER-α36阳性BGC-823细胞的增殖和侵袭能力(均P<0.01)。miR-206通过直接结合CDK14 mRNA的3’-UTR发挥抑制作用,从而负向调节CDK14的表达,进而抑制ER-α阳性BGC-823细胞的增殖和侵袭能力(均P<0.01)。结论:miR-206通过靶向CDK14从而抑制雌激素诱导的ER-α36阳性GC细胞的增殖和侵袭。
Objective:To explore the effects of miR-206 on the proliferation and invasion of ER-α36-positive gastric cancer(GC)BGC-823 cells induced by estrogen and its related mechanisms.Methods:ER-α36-positive BGC-823 cells were stimulated with estradiol(E_(2))at different concentrations(1,10 and 100 pmol/L),then,the expression level of miR-206 was detected by qPCR,the proliferation and invasion were respectively determined by MTT and Transwell assays,and the expression of cyclin-dependent kinase14(CDK14)protein was detected by WB.After transfecting miR-206 mimic,miR-NC,pcDNA-CDK14 or pcDNA-vector into ER-α36 positive BGC-823 cells and stimulating them with 100 pmol/L E_(2),the proliferation and invasion ability of the cells were respectively detected by MTT assay and Transwell assays,and the expression of CDK14 was detected by WB assay.The targeting relationship between miR-206 and CDK14 was verified by Dual luciferase reporter gene assay.Results:E_(2)significantly decreased the expression level of miR-206(P<0.05 or P<0.01),enhanced the proliferation and invasion(P<0.05 or P<0.01),and up-regulated the expression level of CDK14(P<0.01)in ER-α36-positive BGC-823 cells.Overexpression of miR-206 could significantly reduce the proliferation and invasion ability of ER-α36-positive BGC-823 cells induced by E_(2)(all P<0.01).miR-206 negatively regulated the expression of CDK14 by directly binding to the 3’-UTR of CDK14 m RNA,thereby inhibiting the proliferation and invasion of ER-α-positive BGC-823 cells(all P<0.01).Conclusion:miR-206 inhibits the proliferation and invasion of estrogen-induced ER-α36-positive GC cells by targeting CDK14.
作者
杜文凯
张逸寅
杜薇
DU Wenkai;ZHANG Yiyin;DU Wei(Department of Oncology,Huaibei People's Hospital of Anhui Province,Huaibei 235000,Anhui,China;Department of Oncology,Gaoxin Branch of the First Affiliated Hospital of Anhui Medical University,Hefei 230022,Anhui,China;Fifth Department of Internal Medicine,Suixi County Hospital of Traditional Chinese Medicine,Huaibei 235000,Anhui,China)
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2021年第7期689-695,共7页
Chinese Journal of Cancer Biotherapy
基金
安徽省自然科学基金资助项目(No.1908085QH333)。
