ARID1A对乳腺癌生长的抑制及潜在调控机制  

ARID1Aacts as a tumor-suppressor in breast cancer and its underlying regulatory mechanism

作　　者：张倩[1] 颜海波[1] 刘锋[1] 

机构地区：[1]复旦大学生物医学研究院医学系统生物学研究中心,上海200032

出　　处：《复旦学报（医学版）》2015年第4期427-434,共8页Fudan University Journal of Medical Sciences

基　　金：国家重点基础研究发展计划(2011CB910702;2013CB911202);上海市自然科学基金(14ZR1402100)~~

摘　　要：目的探讨ARID1A(AT-rich interactive domain containing protein 1A)对乳腺癌发生、发展的影响,包括对细胞增殖、细胞周期、细胞凋亡的影响及潜在的机制。方法采用小发夹RNA(short hairpin RNA,shRNA)及小干扰RNA(small interfering RNA,siRNA)的RNA干扰技术(RNA interference,RNAi)下调细胞ARID1A基因表达,采用瞬时表达及稳定表达方法过表达ARID1A;通过Western blot检测干扰或过表达ARID1A基因后其蛋白表达水平及P-Akt、PARP和Caspase-3蛋白水平的变化,应用MTT法、细胞计数法及克隆形成检测细胞增殖的变化;应用流式细胞仪检测细胞凋亡及细胞周期的变化。结果应用RNA干扰技术沉默ARID1A基因后,乳腺癌细胞或正常乳腺细胞增殖速度显著上升(Bcap-37-siARID1A,P<0.001;MCF7-siARID1A,P<0.01;MCF10A-shARID1A,P<0.001;HMEC-shARID1A,P<0.001),Akt磷酸化水平上升,克隆形成能力显著提高。过表达ARID1A后,细胞增殖速率均显著下降(P<0.001),Akt磷酸化水平下降,细胞周期发生变化,其中S期减少,G2期增多,同时细胞凋亡比例上升。结论 ARID1A在乳腺癌中发挥着抑制肿瘤生长的作用,提示ARID1A可能成为临床检测、诊断和治疗的有效潜在靶标。Objective To investigate the functional role and the underlying mechanism of AT-rich interactive domain containing protein 1A （ARID1A） in breast cancer,including its influence on cell proliferation,cell cycle and cell apoptosis in vitro. Methods ARID1A was down-regulated by RNA interference （RNAi） of short hairpin RNA （shRNA） and small interfering RNA （siRNA） or overexpressed by transient or stable transfection. The protein changes of P-Akt,PARP and Caspase-3 after ARID1A silencing or overexpression were determined by Western blot. Ceil proliferation was measured using the MTT method and cell counting, while tumorigenicity was detected by colony formation assay. Furthermore, changes in cell cycle and cell apoptosis were measured by FACS. Results Breast cancer or normal breast cells with ARIDI A silencing showed an elevated proliferation rate （Bcap-37-siARID1A, P〈0. 001 ; MCF7-siARID1A, P〈0. 01 ; MCF10A-shARID1A, P〈0. 001 HMEC - shARID 1 A, P 〈 0. 001）, induced phosphorylation of Akt and an increased colony formationrate. Over expression of ARID1A could down-regulate cell proliferation （P〈0. 001）, and also down-regulate Akt phosphorylation. Expression of ARID1A also induced cell accumulation in G2-phase,eeli decrease in S-phase, and cell apoptosis obviously. Conclusions As a tumor suppressor in breast cancer. ARID1A may be a potential clinicall biomarker in detection,diagnosis and therapy.

关 键 词：ARID1A 肿瘤抑癌基因 乳腺癌 

分 类 号：R737[医药卫生—肿瘤]