Reversion of multidrug resistance in leukemia K562 cells by RNA interference targeting homeobox A10 gene / 肿瘤

ABSTRACT

Objective: To investigate a new strategy to reverse multidrug resistance of chronic myeloid leukemia cell line K562 by RNA interference technique through constructing an eukaryotic vector of short hairpin RNA (shRNA) targeting homeobox A10 (HOXA10) gene. Methods: The eukaryotic vector pGPHI/GFP/Neo-HOXA10 with shRNA targeting HOXA10 gene was constructed and then transfected into K562 cells by positive ion liposome. The stable transfectants were vertificated by reverse transcriptase PCR (RT-PCR) 4 weeks after G418 pressure selection. The changes of sensitivity of K562 cells to leurocristine (VCR) and etoposide (VP-16) after transfection with shRNA-HOXA10 were detected by MTT method, and the apoptosis rate was detected by flow cytometry. Results: After selection with G418, the cell clones stably transfected with pGPHI/GFP/Neo-HOXA10 were successfully constructed and verificated by RTPCR. The half inhibitory concentration (IC50) values of VCR and VP-16 for K562 cells transfected with shRNA-HOXA10 were significantly reduced and the apoptosis rate of K562 cells after HOXA10 gene interference combining with chemotherapy was increased significantly as compared with those of shRNA-negative control group and the normal control group (P0.05). Conclusion: The eukaryotic vector of short hairpin RNA (shRNA) targeting HOXA10 gene can enhance the abilities of VCR and VP-16 to inhibit the cell proliferation and induce the apoptosis of K562 cells, namely an increase of sensitivity to these chemotherapeutics in K562 cells. It is hinted that RNA interference targeting HOXA10 gene may reverse the multidrug resistance of leukemia cells in some degree. Copyright© 2011 by TUMOR.