miR-1246对人宫颈癌SiHa细胞增殖、侵袭、迁移能力的影响及其靶基因的初步研究 认领 被引量:7

Effects of miR-1246 on proliferation,invasion and migration of cervical squamous cell carcinoma cell line SiHa and its target gene
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摘要 目的:探讨miR-1246对人宫颈癌 SiHa细胞增殖、侵袭、迁移能力的影响。方法将 SiHa细胞分成3组,分别转染miR-1246 mimics(模拟物)、miR-1246 inhibitor(抑制物)、空白质粒,并测定转染效率。采用 MTT法对比转染后SiHa细胞增殖能力的区别;Transwell小室、划痕实验对比转染后 SiHa 细胞侵袭、迁移能力的区别;Western blot 对比转染后 SiHa细胞表达THBS2(血小板反应蛋白2)的区别。构建THBS2的3’UTR双荧光素酶质粒,与miR-1246共转染入 SiHa细胞,检测荧光素酶的相对活性。结果转染 miR-1246 mimics 的 SiHa细胞,MTT 试验显示其增殖能力较另外两组强;划痕实验显示细胞的迁移能力较另两组强;迁移、侵袭实验显示该组细胞穿膜数量较另外两组多(P<0.01);WB 实验显示其 THBS2蛋白低表达(灰度值6.28±10.22,与空白对照组相比P=0.013)。转染 miR-1246 inhibitor的 SiHa细胞,MTT试验显示其生长速度较另外两组慢;划痕实验显示细胞的迁移能力较另两组弱;迁移、侵袭实验显示该组细胞穿膜数量较另外两组少(P<0.01);WB实验显示其 THBS2蛋白表达稍高(灰度值12.90±19.81,与空白对照组相比P=0.037)。共转染 miR-1246 mimics 和包含 THBS2的3’UTR 端双萤光素酶质粒后, SiHa细胞荧光素酶表达降低。结论 miR-1246对人宫颈癌 SiHa 细胞的增殖、侵袭、迁移能力有促进作用,THBS2是其靶基因,降调靶蛋白THBS2的表达,可能是miR-1246促进宫颈癌发生发展的其中一个机制。 Objective To explore the effects of miRNA-1246 (miR-1246)on cell proliferation,invasion and migration in human cervical squamous cell carcinoma (CSCC)cell line SiHa.Methods SiHa cells were assigned into 3 groups:miR-1246 analog group,miR-1246 antagonist group and control group.Transfection efficiency was determined.The MTT assay,transwell assay and wound healing assay were performed respectively to evaluate the proliferation,invasion and migration abilities of SiHa cells.Western blot was carried out to detect the expression of thrombospondin-2 (THBS2)before and after transfection.A THBS2 3’-UTR-containing dual luciferase plasmid was synthesized and co-transfected with miR-1246 into SiHa cells to observe the luciferase enzyme activity.Results MTT assay,transwell assay and wound healing assay revealed that the abilities of proliferation,migration and invasion were significantly enhanced (P〈0.01)in SiHa cells transfected with miR-1246 analog,but suppressed in SiHa cells transfected with miR-1246 antagonist.Western blot showed that SiHa cells transfected with miR-1246 analog had significantly decreased THBS2 expression (gray value = 6 .2 8 ± 1 0 .2 2 , P=0 .0 1 3 ) while those transfected with miR-1246 antagonist had significantly increased THBS2 expression (gray value = 12.90±19.81, P=0.037).After co-transfected with miR-1246 and THBS2 3’-UTR-containing plasmid,SiHa cells exhibited a decreased level of luciferase enzyme expression.Conclusion miR-1246 promoted the proliferation,invasion and migration of CSCC SiHa cell, and it might promote CSCC tumorigenesis and progression by suppressing the expression of its target gene THBS2 .
作者 陈军莹 姚德生 贺婵娟 丁楠 赵珊 李力 龙凤宜 CHEN Jun-ying, YAO De-sheng, HE Chan-juan, DING Nan, ZHAO Shan, LI Li, LONG Feng-yi Department of Gynecology, the First Affiliated Hospital of Guangxi Medical University 2. Department of Gynecological Oncology, the Affiliated Cancer Hospital of Guangxi Medical University, Nanning 530000, China)
出处 《西安交通大学学报:医学版》 CAS CSCD 北大核心 2015年第2期195-200,共6页 Journal of Xi‘an Jiaotong University:Medical Sciences
基金 广西自然科学基金(2011GXNSFA018184,2013GXNSFBA019130,2013GXNSFBA019132) 广西卫生厅自筹课题(z2012071)
关键词 宫颈癌 微小RNA-1246 血小板反应蛋白2 SIHA细胞 增殖 侵袭 转移 cervical cell carcinoma miRNA- 1246 ( miR- 1246) thrombospondin-2 (THBS2) SiHa cell cellproliferation cell invasion cell migration
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