三氧化二砷抑制人肝癌细胞P27^(kip1)187位苏氨酸磷酸化  

Arsenic trioxide inhibits phosphorylation of P27^(kip1) threonine residue 187 in human hepatic carcinoma cells

作　　者：王酉[1,2] 陆牡丹[1] 李鹏[3] 崔小鹏[3] 沈爱国[1] 

机构地区：[1]南通大学微生物与免疫学教研室,江苏南通226001 [2]无锡市妇幼保健院,江苏无锡214002 [3]南通大学附属医院普外科,江苏南通226001

出　　处：《基础医学与临床》2008年第10期1009-1014,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(30300099)(30770488);江苏省自然科学基金(BK2003035);江苏省高校自然科学研究项目(03KJB180109;04KJB320114);江苏省科技指导性计划(BS2004526);"六大人才高峰"第二批项目资助计划;江苏省卫生科研项目(H200632);南通社会发展科技项目(No.S2007041);美国中华医学基金会基金(06-837)

摘　　要：目的探讨三氧化二砷(As2O3)对人肝癌SMMC-7721细胞的生长抑制与P27kip1第187位苏氨酸(P27Thr187)磷酸化的关系。方法体外培养人肝癌细胞株SMMC-7721,2μmol/L As2O3处理72 h,细胞计数法检测SMMC-7721生长,流式细胞仪检测细胞周期,核质分离、Western Blot及细胞免疫荧光技术检测P27、Thr187磷酸化的P27(p-P27T187)在SMMC-7721细胞中的表达及亚细胞定位。结果2μmol/L As2O3明显抑制SMMC-7721的增殖,细胞周期阻滞在G2/M期。As2O3作用后P27蛋白总量增加,p-P27T187蛋白总量减少,并伴有Cdk2及cyclinE表达下降,同时P27发生从胞质向胞核的易位,p-P27T187核内表达减少。结论As2O3可抑制P27T187的磷酸化,诱导P27在SMMC-7721细胞核中的积聚。Objective To investigate the relationship between growth inhibiting effect of arsenic trioxide （ As2O3 ） and phosphorylation of P27^kip1 threonine residue 187 （P27T187 ） in human hepatocellular carcinoma（HCC） cell line SMMC-7721. Methods SMMC-7721 were treated for 72 h with 2 μmol/L As2O3. The cell growth inhibition was detected by cell counting and the cell cycle was detected by flow cytometry（FCM）. The expression and localization of P27, T187 phosphorylated P27（p-P27T187） were detected by Subcellular Fractionation, Western blot and immunoflurescence. Results As2O3 significantly inhibited the proliferation of SMMC-7721 cell and cell cycle was arrested in Gz/M. A significant decrease in p-P27T187 expression and a reciprocal increase in P27 expression were found in 2 μmol/L As2 03 -treated SMMC-7721 cell. Meanwhile, As2O3 decreased the protein levels of Cdk2 and cyclinE. The location of P27 was transferred from cytoplasm to nuclei and the expression of p-P27T187 was decreased in nuclei. Conclusion As2O3 inhibits the phosphorylation of P27T187, thereby promoting P27 accumulation in SMMC-7721 cell nuclei, inducing cell cycle arrest and growth inhibition.

关 键 词：三氧化二砷 肝细胞癌 SMMC-7721 P27 磷酸化 

分 类 号：R735.7[医药卫生—肿瘤]