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Role of WAVE1 in drug resistance of K562/A02 leukemia cells

KANG Rui CAO Li-zhi YU Yan HU Ting WANG Zhuo XU Wang-qiong XIE Min Department of Pediatrics,Xiangya Hospital,Central South University,Changsha 410008,China  
Objective To investigate if WAVE1 is involved in mult drug-resistance(MDR)of hu- man leukemia cell line K562/A02.Methods The level of WAVE1 in K562 and K562/A02 cells was as- sayed by Western blot and RT-PCR;K562 cells and K562/A02 cells were transient transfected with pEFBOS- WAVE1 reconstructed plasmid or specifically siRNA to WAVE1.50% inhibition concentration(IC_(50))of doxorubiein on K562/A02 was determined by WST-8 assay.Hoechst33258 staining was used to examine cell morphological changes and to calculate percentage of apoptotic nuclei.The mRNA level of mdr1 was assayed by RT-PCR.The Bcl-2 protein was assayed by Western blot.Results ①The WAVE1 expression at mRNA and protein level in K562/A02 cells was increased by about 70% and 63% respectively as compared with that in K562 cells.②Overexpression of WAVE1 in K562 cells by transient transfection significantly increased the resistance to doxorubiein,and increased IC_(50)from(0.05±0.00)μg/ml to(2.99±0.12)μg/ml,and at 1 μg/ml or 5 μg/ml of doxorubicin treatment,cell apoptotic nuclei rate was decreased by 30% or 35% respec- tively.③Suppression of WAVE1 in K562/A02 cells by siRNA resulted in a reversal of MDR to doxorubicin, and decreased IC_(50)from(4.29±0.15)μg/ml to(1.85±0.07)μg/ml,and at 1 μg/ml or 5 μg/ml of doxorubicin treatment,cell apoptotie nuclei rate was increased by 24% or 21% respectively.④Overexpres- sion of WAVE1 in K562 cells significantly increased the mdr1 mRNA and the Bcl-2 protein,and suppression of WAVE1 in K562/A02 cells by siRNA decreased the mRNA and the protein.Conclusion WAVE1 in- volves in the MDR mechanisms in K562/A02 leukemia cells through regulation the level of mdr1 and Bcl-2.
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