耐As_2O_3的白血病细胞系SHI-1/AS2的建立及耐药机制的初步研究  

Establishment of As_2O_3 Resistant Cell Line,SHI-1/AS2,and the Mechanism of Its Drug Resistance

作　　者：王勇[1] 薛永权[1] 陈苏宁[1] 吴亚芳[1] 潘金兰[1] 张俊[1] 沈娟[1] 

机构地区：[1]苏州大学附属第一医院江苏省血液研究所,卫生部血栓与止血重点实验室,江苏苏州215006

出　　处：《苏州大学学报（医学版）》2009年第4期597-600,603,F0002,共6页Suzhou University Journal of Medical Science

基　　金：国家自然科学基金资助项目(30370596);江苏省135重点学科开放课题基金资助项目(135XYO414)

摘　　要：目的建立三氧化二砷(As2O3)的耐药细胞株,进一步从分子水平研究其耐药机制。方法采用高度短时间接触培养方法建立耐As2O3的白血病细胞系(SHI-1/AS2)。细胞倍增时间和耐药倍数测定采用MTT法;细胞周期和细胞免疫表型测定采用流式细胞分析术;常见的10个耐药基因检测和比较采用实时定量PCR(RQ-PCR)。结果耐As2O3的白血病细胞系SHI-1/AS2的细胞倍增时间与SHI-1相似,差异无统计学意义(P>0.05)。SHI-1/AS2细胞G0/G1期细胞少于SHI-1细胞,而G2/M、S期细胞则多于SHI-1细胞,差异均有高度统计学意义(均P<0.01)。SHI-1/AS2细胞对As2O3的耐药倍数是SHI-1的2.7倍,差异有统计学意义(P<0.01)。SHI-1/AS2细胞的集落形成率高于SHI-1细胞。SHI-1/AS2细胞的耐药相关基因Bcl-2有明显上调,Bax、Fas及MGMT下调,而其他耐药相关基因无明显变化。结论建立一株对As2O3产生耐药的白血病细胞系SHI-1/AS2,耐药倍数是SHI-1的2.7倍,其耐药机制与Bcl-2、Bax和Fas基因介导的细胞凋亡调控有关。Objective To establish the As2O3 resistant cell line SHI-1/AS2, and to investigate the mechanism of SHI-1/AS2＇s resistance to As203. Methods The resistance of SHI-1 to As203 gradually was induced by short-time co-culturing SHI-1 with As203. The cell doubling time and the resistant capa- bility to As203 of SHI-1 and SHI-1/AS2 were evaluated by thizolyl blue method（MMT）. The immunopro- file and the cell cycle distribution of them were studied by flow cytometry （FCM）. The ten drug resis- tance-related genes were tested by RQ-PCR. Results There was no significant difference between the cell doubling time of SHI-1 and SHI-1/AS2 （/9〉0.05）. To compare with SHI-1, the portion of cells in Go/ G1 phase of SHI-1/AS2 decreased, but increased in GJM phase （P〈0.01）. SHI-1/AS2＇s colony forming a- bility increased and the resistant capability to As203 was 2.7 times more than that of SHI-1 （P〈0.O1）. In SHI-1/AS2 cells, drug resistance-related gene Bel-2 was up-regulated; Bax, Fas and MGMT were down- regulated, while there was no apparent change for other genes. Conclusion SHI-1/AS2 has the resis- tant capability to As203 to a certain degree, and the resistant capability to As203 is 2.7 times more than that of SHI-1; Apoptosis regulation is involved in drug resistance SHI-1/AS2 cell line, which is mediated by drug resistance-related gene （Bel-2, Bax and Fas）.

关 键 词：抗药性 砷剂 白血病 

分 类 号：R733.7[医药卫生—肿瘤]