长链非编码RNA ATB对三阴性乳腺癌细胞的增殖、凋亡及上皮间充质转化的影响  

Effect of long non-coding RNA ATB on proliferation,apoptosis and epithelial mesenchymal transition of triple negative breast cancer cells

作　　者：郭晨旭[1] 刘静波 许睿[1] 朱超[1] 张立功[1] 钱军[1] GUO Chenxu;LIU Jingbo;XU Rui;ZHU Chao;ZHANG Ligong;QIAN Jun(Department of Surgical Oncology,First Affiliated Hospital of Bengbu Medical College,Bengbu 233000,China;Department of Gynecological Oncology,First Affiliated Hospital of Bengbu Medical College,Bengbu 233000,China)

机构地区：[1]蚌埠医学院第一附属医院肿瘤外科,蚌埠233000 [2]蚌埠医学院第一附属医院肿瘤妇科,蚌埠233000

出　　处：《山西医科大学学报》2022年第8期950-956,共7页Journal of Shanxi Medical University

基　　金：蚌埠医学院自然重点项目(2020byzd053);安徽省教育厅重点项目(KJ2021A0816)。

摘　　要：目的探索三阴性乳腺癌细胞(triple negative breast cancer,TNBC)中长链非编码RNA ATB(long non-coding RNA activated by transforming growth factorβ,lncRNA ATB)对其增殖、凋亡、侵袭及上皮间充质转化(epithelial-mesenchymal transition,EMT)的作用。方法收集2020年1月至2020年12月蚌埠医学院第一附属医院肿瘤外科行手术治疗的18例TNBC患者的癌组织,以对应的癌旁组织作为对照。qRT-PCR检测lncRNA ATB在TNBC癌组织及癌旁组织中的表达。qRT-PCR检测lncRNA ATB在TNBC细胞MDA-MB-231与人乳腺上皮细胞MCF-10A中的表达情况。下调MDA-MB-231细胞中lncRNA ATB的表达为si-ATB组,并检测沉默效率,阴性小干扰RNA作对照,为si-NC组。CCK-8、流式细胞仪、划痕实验、Transwell实验分别检测下调lncRNA ATB对MDA-MB-231细胞增殖、凋亡、迁移、侵袭能力的影响。Western blot检测下调lncRNA ATB对MDA-MB-231细胞中上皮间充质转化(epithelial-mesenchymal transition,EMT)相关蛋白E-cadherin、vimentin、N-cadherin的影响。结果相比癌旁组织,lncRNA ATB在TNBC组织中的表达明显增加(P<0.05)。相比人乳腺上皮细胞MCF-10A,TNBC细胞MDA-MB-231中lncRNA ATB的表达明显增加(P<0.05)。CCK-8结果显示相比si-NC组,si-ATB组MDA-MB-231细胞的增殖能力在48,72,96 h时明显减弱(均P<0.05)。流式细胞仪结果显示相比si-NC组,si-ATB组MDA-MB-231细胞的凋亡率明显增加(12.24%±1.52%vs 24.33%±2.15%,P<0.05)。相比si-NC组,划痕实验结果显示si-ATB组MDA-MB-231细胞的迁移能力明显减弱(61.0%±5.0%vs 31.7%±3.3%,P<0.05)。相比si-NC组,Transwell实验结果显示si-ATB组MDA-MB-231细胞的侵袭能力明显减弱(40.33±3.51 vs 15.67±2.52,P<0.05)。Western blot结果显示,相比si-NC组,si-ATB组E-cadherin蛋白在MDA-MB-231细胞中的相对表达量明显增加(0.42±0.04 vs 0.63±0.05,P<0.05);相比si-NC组,si-ATB组vimentin蛋白在MDA-MB-231细胞中的相对表达量明显减弱(0.55±0.05 vs 0.32±0.04,P<0.05);相比si-NC组,si-ATB组Objective To explore the effect of long non-coding RNA ATB(long non-coding RNA activated by transforming growth factorβ,lncRNA ATB)on proliferation,apoptosis,invasion and epithelial-mesenchymal transition(EMT)of triple negative breast cancer(TNBC).Methods The cancerous tissues of 18 patients with TNBC undergoing surgery in the department of surgical oncology in the First Affiliated Hospital of Bengbu Medical College from January 2020 to December 2020 were collected,and the corresponding paracancerous tissues(normal tissues)were used as controls.The expression of lncRNA ATB in TNBC cancerous tissues and paracancerous tissues was detected by qRT-PCR.The expression of lncRNA ATB expression in TNBC cells MDA-MB-231 and human mammary epithelial cells MCF-10A was detected by qRT-PCR.The expression of lncRNA ATB in MDA-MB-231 cells was inhibited as si-ATB group,and negative siRNA was used as control(si-NC group),and then the infection efficiency was detected.CCK-8 was used to detect the effect of lncRNA ATB on the proliferation of MDA-MB-231.The effect of lncRNA ATB on MDA-MB-231 apoptosis was detected by flow cytometry.The effects of lncRNA ATB on the migration and invasion of MDA-MB-231 cells were detected by scratch test and Transwell test.The effects of lncRNA ATB on E-cadherin,vimentin and N-cadherin in MDA-MB-231 were detected by Western blot.Results The expression of lncRNA ATB in tissues and cells of TNBC was significantly higher than that of normal tissues and MCF-10A cells,respectively(P<0.05).Compared with si-NC group,the results of CCK-8 showed that the proliferation of MDA-MB-231 cells in si-ATB group was significantly reduced at 48 h,72 h and 96 h(all P<0.05).Compared with si-NC group,the results of flow cytometry showed that the apoptosis rate of MDA-MB-231 cells in si-ATB group was significantly increased(12.24%±1.52%vs 24.33%±2.15%,P<0.05).Compared with si-NC group,the results of scratch test showed that the migration ability of MDA-MB-231 cells in si-ATB group was significantly reduced(61.0%±5.0%vs

关 键 词：lncRNA ATB 三阴性乳腺癌 上皮间充质转化 增殖 凋亡 

分 类 号：R737.9[医药卫生—肿瘤]