核糖体蛋白L6对K562-A02细胞耐药性及凋亡影响.docVIP

核糖体蛋白L6对K562/A02细胞耐药性及凋亡影响 　 作者：陈宏 谢兆霞　姜浩 张志伟 王光平 【摘要】 　　本研究观察核糖体蛋白L6(RPL6)基因表达的改变对白血病细胞耐药性的作用及其可能的机制。通过RTPCR 方法获得RPL6cDNA序列，用真核表达载体pcDNA3.1（+）分别构建正向插入和反向插入的RPL6 cDNA 重组质粒。以脂质体将正义RPL6 cDNA 真核表达质粒转染K562细胞,将反义RPL6 cDNA真核表达质粒转染K562/AO2细胞。以MTT、流式细胞术和荧光分光光度计观察RPL6对化疗药物耐药性、凋亡和caspase3的作用。结果表明：　转染正义RPL6cDNA 真核表达质粒后,K562细胞对阿霉素的耐药性增强到原来的325%，凋亡和caspase3活性明显降低（Plt;0.005）；转染反义RPL6 cDNA 真核表达质粒后,K562/A02细胞对阿霉素的耐药性降低原来的38%；凋亡和caspase3活性明显增加（Plt;0.005）。结论：　RPL6基因过表达通过改变药物诱导的凋亡在K562/A02细胞耐药性的形成中起重要作用。 【关键词】 核糖体蛋白L6； 　抗药性；　细胞株　 K562/A02 　　Effect of Ribosomal Protein L6 on Drug Resistance and Apoptosis in K562/A02 Cells Abstract The objective of study was to investigate the effect of ribosomal protein L6 (RPL6) gene expression on the drug resistance of leukemia cells and its possible mechanism. RPL6 cDNA was obtained by RTPCR, both sense and antisense cDNA recombinants of RPL6encoding gene were constructed with pcDNA3. 1(+) expression vector. Subsequently , the sense RPL6 cDNA recombinant was transfected into K562 cells while the antisense one into K562/A02 cells by liposomal reagent. The chemosensitivity, apoptosis and caspase3 activity of K562 and K562/A02 cells were evaluated by MTT assay, flow cytometer and fluorometer respectively. The results indicated that expression of RPL6 in K562/A02 was higher than that in K562; resistance of sensetransfected K562 cells to doxorubicin was 325% of control cells, apoptosis and caspase3 activity decreased (Plt;0.005); whereas resistance of antisensetransfected K562/A02 cells to adriamycin was 38% of control cells, apoptosis and caspase3 activity significantly increased (Plt;0.005). It is concluded that RPL6 gene plays an important role in the development of drug resistance in K562/A02 cells by changing druginduced apoptosis. Key words Ribosomal protein L6; 　Drug resistance; 　Cell line; K562/A02 J Exp Hematol 2007; 15(2):292-295 白血病生存期短，病死率高，其主要原因之一是多药耐药（multidrug resistance， MDR）的产生。MDR 机制复杂，如以P糖蛋白为代表的膜泵蛋白的表达、细胞解毒能力的增强、DNA