miR-30c抑制宫颈癌细胞恶性表型的分子机制研究  

Mechanism of miR-30c over-expression inhibiting malignant phenotypes of cervical cancer cells

作　　者：金虹[1] 张萌[1] 刘念[1] 李珊[1] JIN Hong;ZHANG Meng;LIU Nian;LI Shan(Department of Obstetrics, Center for Maternal-Fetal Medicine, The First Affiliated Hospital of Xinjiang Medical University, Wulumuqi 830011, China)

机构地区：[1]新疆医科大学第一附属医院母胎医学中心产科,新疆乌鲁木齐830011

出　　处：《中国病理生理杂志》2018年第5期804-811,共8页Chinese Journal of Pathophysiology

摘　　要：目的:探讨微小RNA(miR)-30c过表达抑制宫颈癌细胞恶性表型的分子机制。方法:运用Lipofectamine 2000转染法将pGenesil-1-miR-30 c质粒转染宫颈癌细胞系C33A、HeLa、SiHa和CaSki,以转染p-Geresil-1的细胞为阴性对照;运用TaqMan real-time PCR检测各组细胞中miR-30c的表达水平;采用MTT法、集落形成实验、Transwell法、Annexin V-FITC及流式细胞术分别测定细胞活力抑制率、集落形成能力、迁移率及凋亡率;Western blot检测Bax、Bcl-2、基质金属蛋白酶(MMP)-9、MMP-13和金属蛋白酶组织抑制物1(TIMP-1)的蛋白表达水平。结果:转染p Genesil-1-miR-30c质粒的宫颈癌细胞系中miR-30c表达水平均显著高于阴性对照组(P<0.01);过表达miR-30 c的宫颈癌细胞活力抑制率显著高于阴性对照组(P<0.05),细胞集落形成率和迁移率显著低于阴性对照组(P<0.05);流式细胞术检测结果显示,miR-30c过表达的宫颈癌细胞凋亡率显著高于对照组(P<0.05);Western blot结果显示miR-30 c过表达促进Bax和TIMP-1蛋白的表达,而抑制Bcl-2和MMP-13蛋白的表达(P<0.05或P<0.01)。结论 :miR-30 c过表达抑制宫颈癌细胞活力和迁移,诱导宫颈癌细胞凋亡,其机制与激活细胞凋亡通路及抑制MMP-13表达有关。AIM： To investigate the molecule mechanism of microRNA（ miR）-30 c over-expression inhibiting malignant phenotypes of cervical cancer cells. METHODS： Cervical cancer cell lines C33A,HeLa,SiHa and CaSki were transfected with pGenesil-1-miR-30 c plasmid using Lipofectamine 2000 kit,and the expression of miR-30 c was determined by Taq Man real-time PCR. The cell viability inhibition rate,colony formation ability,migration rate and apoptotic rate were measured by MTT assay,colony formation assay,Transwell experiment,and flow cytometry with Annexin V-FITC staining.The protein expression of Bax,Bcl-2,matrix metalloproteinase（ MMP）-9,MMP-13 and tissue inhibitor of metalloproteinase-1（ TIMP-1） was detected by Western blot. RESULTS： The expression levels of miRNA-30c in the cervical cancer cell lines transfected with p Genesil-1-miR-30c plasmid were significantly higher than those in negative control groups（ cell lines transfected with p Genesil-1 plasmid）（ P〈0. 01）. Significantly increased cell viability inhibition rate,and decreased colony formation ability and migration rate were found in the cervical cancer cell lines over-expressing miR-30c as compared with negative control groups（ P〈0. 05）. The apoptotic rate in the cervical cancer cell lines over-expressing miR-30c was dramatically higher than that in control groups（ P〈0. 05）. Over-expression of miR-30c in cervical cancer cells promoted the protein expression of Bax and TIMP-1,and decreased the protein expression of Bcl-2 and MMP-13（ P〈0. 05 or P〈0. 01）. CONCLUSION： Over-expression of miR-30c significantly inhibits the viability and migration,and induces apoptosis of cervical cancer cells. The mechanism may be related to activating apoptosis pathway and inhibiting MMP-13 protein expression.

关 键 词：宫颈癌 微小RNA-30c 细胞活力 细胞凋亡 基质金属蛋白酶 

分 类 号：R737.33[医药卫生—肿瘤] R363[医药卫生—临床医学]