CAJ | 학술논문

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。
목적：탐토miR-1246대인궁경암 SiHa세포증식、침습、천이능력적영향。방법장 SiHa세포분성3조,분별전염miR-1246 mimics(모의물)、miR-1246 inhibitor(억제물)、공백질립,병측정전염효솔。채용 MTT법대비전염후SiHa세포증식능력적구별；Transwell소실、화흔실험대비전염후 SiHa 세포침습、천이능력적구별；Western blot 대비전염후 SiHa세포표체THBS2(혈소판반응단백2)적구별。구건THBS2적3’UTR쌍형광소매질립,여miR-1246공전염입 SiHa세포,검측형광소매적상대활성。결과전염 miR-1246 mimics 적 SiHa세포,MTT 시험현시기증식능력교령외량조강；화흔실험현시세포적천이능력교령량조강；천이、침습실험현시해조세포천막수량교령외량조다(P<0.01)；WB 실험현시기 THBS2단백저표체(회도치6.28±10.22,여공백대조조상비P=0.013)。전염 miR-1246 inhibitor적 SiHa세포,MTT시험현시기생장속도교령외량조만；화흔실험현시세포적천이능력교령량조약；천이、침습실험현시해조세포천막수량교령외량조소(P<0.01)；WB실험현시기 THBS2단백표체초고(회도치12.90±19.81,여공백대조조상비P=0.037)。공전염 miR-1246 mimics 화포함 THBS2적3’UTR 단쌍형광소매질립후, SiHa세포형광소매표체강저。결론 miR-1246대인궁경암 SiHa 세포적증식、침습、천이능력유촉진작용,THBS2시기파기인,강조파단백THBS2적표체,가능시miR-1246촉진궁경암발생발전적기중일개궤제。
ABSTRACT: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 .