Peng Zhi, Xiao Zhi-jian, Wang Yi, Liu Peng, Cai Ying-lin, Feng Wen-li, Han Zhong-chao
State Key Laboratory of Experimental Hematology, Institute of Hematology, CAMS & PUMC, Tianjin 300020, China.
Zhonghua Xue Ye Xue Za Zhi. 2004 Jan;25(1):5-7.
To investigate the effect of small interference RNA (siRNA) on mdr1 and P-glyco-protein (P-gp) expression of multi-drug resistance (MDR) human leukemia cell line K562/A02.
Three si RNAs (si-mdr1-1, si-mdr1-2, si-mdr1-3) which were specifically targeted mdr1 gene were synthesized and transfected into K562/A02 cells. Expression of mdr1 mRNA was assayed by RT-PCR. P-gp expression and intracellular daunorubicin (DNR) concentration were determined by flow cytometry. 50% inhibition concentration (IC(50)) of doxorubicin (ADM) on K562/A02 was determined by MTT method.
Treatment of K562/A02 cell with the 3 kinds of siRNAs resulted in a reversal of MDR of a different extent. The third siRNA was more effective in the suppression of mdr1 with a significant reduction of (58.0 +/- 1.54)% of the mdr1 mRNA expression. Positive expression rate of p170 decreased from (76.0 +/- 1.0)% to (19.6 +/- 1.9)%, and the relative efficiency of K562/A02 to ADM was 70.4%. The intracellular accumulation of DNR increased after siRNA treatment.
The siRNA could effectively restore the sensitivity of K562/A02 cells to conventional chemotherapeutic agents.
研究小干扰RNA（siRNA）对多药耐药（MDR）人白血病细胞系K562/A02中mdr1和P-糖蛋白（P-gp）表达的影响。
合成3种特异性靶向mdr1基因的siRNA（si-mdr1-1、si-mdr1-2、si-mdr1-3），并转染至K562/A02细胞。采用逆转录聚合酶链反应（RT-PCR）检测mdr1 mRNA的表达。通过流式细胞术测定P-gp表达及细胞内柔红霉素（DNR）浓度。采用噻唑蓝（MTT）法测定阿霉素（ADM）对K562/A02的半数抑制浓度（IC50）。
用3种siRNA处理K562/A02细胞后，不同程度地逆转了多药耐药。第3种siRNA对mdr1的抑制效果更佳，mdr1 mRNA表达显著降低（58.0±1.54）%。p170阳性表达率从（76.0±1.0）%降至（19.6±1.9）%，K562/A02对ADM的相对耐药倍数为70.4倍。siRNA处理后细胞内DNR蓄积增加。
siRNA可有效恢复K562/A02细胞对传统化疗药物的敏感性。
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