汉防己甲素对K562/A02细胞株SORCIN基因表达的影响  

Influence of Tetrandrine on SORCIN Gene Expression in K562/A02 Cell Line

作　　者：李静[1] 陈宝安[1] 朱敏生[2] 高峰[1] 丁家华[1] 高冲[1] 孙耘玉[1] 程坚[1] 王骏[1] 赵刚[1] 宋慧慧[1] 马燕[1] 鲍文[1] 孙新臣[3] 成红艳[3] 许文林[4] 沈惠玲[4] 

机构地区：[1]东南大学附属中大医院血液科,江苏南京210009 [2]南京大学模式动物研究所,江苏南京210009 [3]东南大学附属中大医院肿瘤科,江苏南京210009 [4]江苏大学附属人民医院血液科,江苏镇江212002

出　　处：《中国实验血液学杂志》2008年第1期65-69,共5页Journal of Experimental Hematology

基　　金：国家自然科学基金资助(编号39970832,30740062);省科技发展计划(社会发展)资助(编号BS20020224)

摘　　要：本研究旨在探讨汉防己甲素(Tet)在逆转K562/A02细胞耐药与耐药相关的可溶性钙结合蛋白(SORCIN)基因表达之间的关系,为Tet的临床应用提供新的理论基础。通过MTT法筛选出所需浓度Tet,与K562/A02细胞株孵育培养48小时后,通过MTT检测Tet对柔红霉素(DNR)细胞毒性的影响,用RT-PCR方法检测SORCIN基因表达的变化,用Western blot方法检测sorcin表达蛋白的变化。结果表明:在所选浓度范围内,Tet可以增强DNR对K562/A02的细胞毒作用,(Tet浓度为0,0.5,1.0,2.0mg/L时DNR和Tet的IC50分别为11.3±0.17,5.15±0.10,3.91±0.06,2.52±0.04mg/L,p<0.01);K562/A02细胞中SORCIN基因高表达,且随着Tet浓度的增加SORCIN的表达先增强后减弱(Tet浓度为0.5mg/L时增强,1.0和2.0mg/L时减弱,p<0.05);K652细胞中sorcin蛋白低表达,K562/A02细胞中sorcin蛋白高表达,且随着Tet浓度的增加先增强后减弱(Tet浓度为0.5mg/L时增强、1.0和2.0mg/L时减弱,p<0.05)。结论:MTT检测显示Tet对K562/A02耐药逆转呈浓度依赖性,Tet可能通过调整SORCIN基因及蛋白的表达参与逆转K562/A02的耐药性,但并不与其耐药逆转作用完全相关。This study was aimed to explore the changes of soluble resistance-related calcium binding protein （sorcin） expression in reversion of multidrug resistance of K562/A02 leukemic cell line with defferent concentrations of tetrandrine （Tet）, so as to provide a new theoretic evidence for clinical application of Tet. The inhibition of K562/A02 cell line by daunorubicin （DNR） was assayed by MTT method. The changes of SORCIN gene expression were assayed by RT-PCR. The changes of SORCIN protein expressed were assayed by Western blot. The results showed that Tet could enhance the cytotoxity of DNR to K562/A02 cells （the IC50 of DNR ＋ Tet was 11.3 ±0.17 mg/L, 5.15 ±0.10 mg/L, 3.91 ±0.06 mg/L, and 2.52 ±0.04 mg/L, when concentrations of Tet were 0 mg/L, 0.5 mg/L, 1.0 mg/L, and 2.0 mg/L respectively）. The gene encoding sorcin was highly expressed in K562/A02 cells, the expression of which was firstly enhanced in Tet concentration 0.5 mg/L, then attenuated in Tet concentration of 1.0, 2.0 mg/L （p 〈 0.05）. Sorcin protein expressed lowly in K562 cells and highly in K562/A02 cells, but the expression of SORCIN protein in K562/A02 cells was enhanced in Tet concentration of 0.5 mg/L, then was attenuated in Tet concentration of 1. 0,2.0 mg/L （p 〈0.05）. It is concluded that the effect of Tet on reversal of K562/A02 drug-resistance shows concentration dependence by MTT assay. Tet reverses multidrug-resistance of K562/A02 cells through regulation of expression of SORCIN gene and protein, but not fully correlates to the reversing effect.

关 键 词：多药耐药 耐药相关性可溶性钙结合蛋白 汉防己甲素 K562/A02细胞株 

分 类 号：R733.7[医药卫生—肿瘤] R979.1[医药卫生—临床医学]