多柔比星诱导白血病细胞耐药产生的机制研究.docVIP

多柔比星诱导白血病细胞耐药产生的机制研究 【摘要】 目的： 观察多柔比星诱导人白血病细胞系K562产生耐药的规律，初步探讨耐药产生的机制。方法： 在设定的浓度梯度和作用时间下，用多柔比星处理K562细胞，采用RTPCR法测定mdr1基因及其他耐药相关基因的表达；运用流式细胞仪检测mdr1基因编码的P糖蛋白（Pgp）的表达水平，用免疫荧光染色检测细胞凋亡相关蛋白Survivin的表达。结果： 多柔比星在体外能诱导K562细胞产生耐药性，随着多柔比星作用浓度的增加和时间的延长，K562中mdr1及Pgp的表达逐渐上调，MRP,Topo Ⅱ,YB1和NFkappaB基因的表达亦有改变，GST则无明显变化，凋亡相关基因Survivin及其编码的蛋白在耐药产生过程中表达亦上调。结论： 多柔比星以剂量和时间依赖性方式诱导K562细胞产生耐药，多种耐药相关基因参与诱导耐药的形成，凋亡相关基因的表达改变亦是其耐药形成机制之一。 【关键词】 多柔比星　K562细胞　多药耐药　mdr－1基因　P糖蛋白 Study on mechanisms of doxorubicininduced multidrug resistance　in leukemia cell line K562 QIN Rujuan， XU Wenlin， ZHOU Leilei，TANG Huarong， SHEN Huiling，WANG Fachun (Department of Central Laboratory，the Affiliated Peopleprime;s Hospital of Jiangsu University，Zhenjiang Jiangsu 212002，China) ［Abstract］ Objective: To investigate doxorubicininduced multidrug resistance (MDR) in human leukemia cell line K562 and then the mechanisms of drug resistance were studied. Methods： K562 cells were treated with doxorubicin at different concentrations and times. The expression of mdr1 and other related genes were examined by reverse transcriptionpolymerase chain reaction (RTPCR). Pglycoprotein (Pgp) was detected by flow cytometry (FCM) and the protein Survivin was analysed by immunofluorescence. Results： Incubation of K562 cells with doxorubicin not only induced functionally mdr1 overexpression, but also increased the expression of other genes including MRP, Topo Ⅱ, YB1, NFkappaB and Survivin. Increasing level of mdr1 mRNA correlated with its corresponding Pglycoprotein level in druginduced cells. Survivin, a member of IAP family proteins, was also high expressed after incubation of K562 cells with doxorubicin. Conclusion： Doxorubicin can induce functional multidrug resistance overexpression in K562 cells in dose and time dependent manner, and the functional multidrug resistance may be due to the abnormal expression of genes MRP, Topo Ⅱ, YB1, NFkappaB and the antiapoptosis gene Survivin. ［Key words］ Doxorubicin