2010•Zhongliu fangzhi yanjiuOpen access

Drug Resistance Reversal of Doxorubicin-resistant Human Leukemia Cell Line K562/ADM through Down-regulating MDR1 Gene

Zheng Yan

Abstract

Objective To explore the effect of shRNA targeted to multidrug resistance gene 1(MDR1)by RNA interference(RNAi)on drug resistance of doxorubicin-resistant human leukemia cell line K562/ADM.Methods The RNAi plasmid of pENTRTM/U6-MDR1,targeting to human MDR1 gene,was transfected into doxorubicin-resistant human leukemia cell line K562/ADM and parental cell line K562.After 48h transfection,MDR1mRNA level was determined by real time RT-PCR.The expression and function of P-gp protein was analyzed by flow cytometry.Drug resistance of cells was detected by MTT assay.Results Compared with non-transfected cells,MDR1mRNA and expression levels and function of P-gp protein in cells of pENTRTM/U6-MDR1 group decreased significantly(P0.05).The resistance on doxorubicin was decreased drmatically(P0.05).Conclusion The resistance on doxorubicin of human doxorubicin-resistant leukemia cells was reversed by MDR1 gene down-regulating.

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Key concepts: K562 cells, Doxorubicin, Transfection, RNA interference, Small hairpin RNA, Multiple drug resistance, Cell culture, Flow cytometry