白血病细胞中NFκB调控MDR1基因、P糖蛋白及逆转耐药的研究.docVIP

白血病细胞中NFκB调控MDR1基因、P糖蛋白及逆转耐药的研究 作者：王漪，刘心，张海涛，余敏，王晖 【摘要】 本 研究 探讨NFκB调控的血液肿瘤细胞抗凋亡机制中是否有mdr1的参与，并通过对NFκB的抑制希望获得血液肿瘤的耐药逆转，为难治性白血病的 治疗 探索一条新的道路。取不同终浓度的NFκB抑制剂PDTC作用的，或者同一浓度作用不同时间后的K562/A02细胞，分别用免疫组织化学加 计算 机图像 分析 仪 方法 半定量检测每1份样本中NFκB的活性和Pgp的表达，以及用反转录多聚酶链反应方法半定量检测每2份样本中mdr1的活性，通过统计学来分析mdr1、Pgp的表达是否与NFκB的活化有关。结果表明：随着NFκB抑制剂PDTC作用浓度的不断升高、作用时间的不断延长，K562/A02细胞NFκB/p65的表达不断降低，Pgp和mdr1 mRNA的表达也逐渐降低，而且三因子之间均有显著相关性。结论： 通过对K562/A02细胞高表达的NFκB的抑制可降低该细胞的mdr1 mRNA和Pgp的表达，从而提高血液肿瘤细胞的放化疗敏感性。 【关键词】 PDTC NFκB Regulating Expression of MDR1 Gene and Pgp to Reverse Drugresistance in Leukemic Cells Abstract This study was purposed to investigate whether the mdr1 participates in antiapoptotic mechanisms of hematologic malignant cells regulated by NFκB and to explore the reversal effect of inhibiting NFκB on drugresistance of hematologic malignant cells so as to search a novel way for treatment of refractory leukemia. NFκB activity and Pgp expression in K562/A02 cells cultured with PDTC in different concentration or different time were detected by immunohistochemistry and computerized picture analysis. The mdr1 was detected by reverse transcriptionpolymerase chain reaction, and the relationship among NFκB , Pgp and mdr1 was analyzed. The results showed that in K562/A02 cells the expression of Pgp and mdr1 were positively correlated with NFκB/p65, which showed the concentration and time dependent manner. It is concluded the mechanism of NFκB regulating antiapoptosis in K562/A02 cells has the correlation with the expression of mdr1 and Pgp. The expression of mdr 1 mRNA and Pgp can be reduced through inhibiting NFκB in K562/A02 cells, so that the sensitivity of chemical therapy can be enchanced on hematologic malignant cells. Key words K562/A02； NFκB； PDTC； PGP； MDR1 J Exp Hematol 2007; 15(5):950-954 白血病的多药耐药性是导致化疗失败、疾病复发及难以根治的主要原因，是 目前 临床上亟待解决的难题之一。新近研究发现mdr1及其产物Pgp高表达是耐药的主要机制。白血病细胞MDR现象就是指白血病细胞mdr1基因及其产物Pgp过度表达导致细胞药物外排增加，降低白血病细胞内化疗药物浓度，白血病细胞由此逃避化疗药物的杀伤而产生耐受现象［1］。因此， 应用