K562/A02细胞核糖体蛋白L6的表达与多药耐药性的实验研究

目的观察核糖体蛋白L6(RPL6)基因表达的改变对白血病多药耐药性的作用。方法通过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RPL6cDNA重组质粒,以脂质体将正义RPL6cDNA真核表达质粒转染K562细胞,将反义RPL6cDNA真核表达质粒转染K562/A02细胞,以MTT法检测细胞对阿霉素(ADM)、长春新碱(VCR)、米托蒽醌(MIT)和依托泊苷(VP16)耐药性的作用。结果RPL6在K562/A02细胞中的表达较K562增高(P<0.001),转染正义RPL6cDNA真核表达质粒后,K562细胞对ADM、VCR、MIT、VP16的耐药性比未转染的K562细胞分别增强3.25、1.95、2.00和3.48倍(P<0.05或0.01),转染反义RPL6cDNA真核表达质粒后,K562/A02细胞对ADM、VCR、MIT、VP16的耐药性比未转染的K562/A02细胞分别降低62%、50%、56%和47%(P<0.05或0.01)。结论RPL6基因过表达在K562/A02细胞耐药性的形成中起重要作用。

Expression and effect of human ribosomal protein L6 on multidrug resistance in K562/A02 cells

Objective:To investigate the relationship between ribosomal protein L6 (RPL6) gene and the multidrug resistance of leukemia cell.Method:RPL6 cDNA was obtained by RT-PCR, Both sense and antisense cDNA recombinants of RPL6-encoding gene were constructed with pcDNA3.1( ) eukaryotic expression vector. The sense RPL6 cDNA recombinant was transfected into K562 cells while the antisense one into K562/A02 cells by liposomal reagent. The chemosensitivity of cells to adriamycin (ADM), Vincristine(VCR), Mitoxantrone (MIT) and Etoposide (VP16) was evaluated by MTT assay.Result:Internal control RT-PCR showed higher expression of RPL6 in K562/A02 than in K562 ( P < 0.001 ); Sense-transfected K562 cells were 3.25 , 1.95 , 2.00 and 3.48 times more resistant to ADM, VCR, MIT and VP16 than control cells did ( P < 0.05 or 0.01 ), whereas resistant to these drugs of antisense-transfected K562/A02 cells decreased by 62%, 50%, 56% and 47% than that of control cells ( P < 0.05 or 0.01 ).Conclusion:RPL6 gene plays an important role in the development of drug resistance in K562/A02 cells.