miRNA-210对人乳腺癌细胞增殖、迁移和侵袭的影响  

Effect of miRNA-210 on proliferation,migration and invasion of human breast cancer cells

作　　者：张楠[1] 李少遊 巩雅宁 任俊宇[2] 田晰晰 董坚[1] 

机构地区：[1]昆明医科大学第一附属医院肿瘤内科,云南昆明650032 [2]昆明医科大学第一附属医院肿瘤科,云南昆明650032

出　　处：《中国肿瘤生物治疗杂志》2013年第3期289-294,共6页Chinese Journal of Cancer Biotherapy

基　　金：云南省科技条件平台建设计划项目基金资助(No.2007DA006);昆明医科大学研究生创新基金(No.2012N06)~~

摘　　要：目的:探讨miRNA-210(miR-210)在人乳腺癌组织中的表达及其对人乳腺癌细胞MDA-MB-231增殖、迁移和侵袭的影响。方法:收集2011年10月至2012年6月期间昆明医科大学第一附属医院20例乳腺癌患者组织标本,real-time PCR检测乳腺癌组织和癌旁组织以及乳腺癌细胞MDA-MB-231和正常乳腺细胞MCF-10a中miR-210的表达。采用Lipofectami-neTM2000将miR-210 inhibitor转染至MDA-MB-231细胞中,通过荧光显微镜检测miR-210的转染效率,MTT和软琼脂克隆形成实验检测MDA-MB-231细胞的增殖,流式细胞术检测细胞周期和凋亡,Transwell法检测细胞的迁移、侵袭能力。结果:miR-210在乳腺癌组织和MDA-MB-231细胞中的表达水平均显著高于癌旁组织和正常乳腺细胞(P<0.01)。miR-210 inhibitor成功转染MDA-MB-231细胞,转染效率为(88.29±2.98)%,转染miR-210 inhibitor后MDA-MB-231细胞的增殖和克隆形成能力明显减弱(P<0.05),被阻滞于G0/G1期的细胞数明显增多[(64.23±3.12)%vs(55.53±0.96)%,P<0.01],凋亡细胞比例也显著增加[(31.90±3.05)%vs(15.98±0.63)%,P<0.01],细胞的迁移[(291.00±43.12)vs(1137.38±83.49)个,P<0.01]、侵袭[(131.63±32.01)vs(647.88±31.20)个,P<0.01]均受到明显抑制。结论:miR-210在乳腺癌组织和细胞中过表达,转染miR-210 inhibitor后乳腺癌细胞MDA-MB-231的增殖、迁移和侵袭能力明显减弱。Objective：To investigate the effect of siRNA（small interference RNA,siRNA） silencing aquaporin-5（AQP-5） expression on the proliferation,apoptosis,and chemosensitivity of human colon cancer HT-29 cells.Methods： A synthetic AQP-5-siRNA sequence was transfected into HT-29 cells,and the inhibition efficiency was detected by Western blotting.Sulphorhodamine B（SRB） assay were used to detect the cell proliferation inhibition rate.Cell apoptosis of HT-29 cells were detected by flow cytometry（FCM）.The activity of caspase-3 in HT-29 cells was measured by spectrophotometry.The mRNA and protein levels of PCNA and P53 in HT-29 cells after AQP-5-siRNA transfection were determined by real-time PCR and Western blotting.5-fluorouracil（5-FU） and cisplatin（DDP） were selected to treat the AQP-5-siRNA-HT-29 cells,and SRB assay was used to detect the cell proliferation inhibition rate.The ＂Q＂ Method of Jin Zhenjun was used to evaluate the interaction of the two drugs.Results： SRB assay showed that compared with NC-HT-29 cells,AQP-5 expression was significantly decreased in AQP-5-siRNA-HT-29 cells（P0.05）.Compared with NC-HT-29 cells,the proliferation inhibition rate was increased significantly in AQP-5-siRNA-HT-29 cells（[9.23±0.51]% vs 0,P0.05）.Flow cytometry analysis showed that compared with NC-HT-29 cells,the cell apoptosis rate increased dramatically in AQP-5-siRNA-HT-29 cells（[10.81±1.32]% vs [0.99±0.18]%,P0.05）.The activities of caspase-3 were also increased in AQP-5-siRNA-HT-29 cells compared with NC-HT-29 cells（[0.19±0.03] vs [0.09±0.01],P0.05）.Real-time PCR and Western blotting results indicated that compared with NC-HT-29 cells,the mRNA and protein levels of PCNA were decreased（P0.05）,simultaneously,the mRNA and protein levels of P53 were increased（P0.05） in AQP-5-siRNA-HT-29 cells.Compared with AQP-5-siRNA-HT-29 cells or 5-FU-HT-29 cells,the proliferation inhibition rate was increased remarkably in AQP-5-siRNA＋5-FU-HT-29 cells（[44.93±2.28]% vs [9.11±0.32]%,

关 键 词：乳腺癌 MDA-MB-231 MIRNA miRNA-210 增殖 迁移 侵袭 

分 类 号：R737.9[医药卫生—肿瘤] R730.54[医药卫生—临床医学]