Shan Xue-Yun, Chen Bao-An, Xia Guo-Hua, Xu Wen-Lin, Ding Jia-Hua, Gao Chong, Sun Yun-Yu, Wang Jun, Cheng Jian, Zhao Gang, Bao Wen, Song Hui-Hui, Gao Feng, Wang Fei, Wang Xue-Mei
Department of Hematology, Zhongda Hospital, Southeast University Clinical Medical College, Nanjing 210009, Jiangsu Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2010 Feb;18(1):90-5.
This study was aimed to investigate the reversal effect of tyrosine kinase inhibitors (TKI) Imatinib and Nilotinib on multidrug-resistant cell line K562/A02. The expression levels of mdr-1 mRNA and bcr-abl mRNA were assayed by RT-PCR. The protein levels of P-glycoprotein (P-gp) and P210 were detected by Western blot. The daunorubicin (DNR) accumulation in K562/A02 cells were analyzed by flow cytometry (FCM). The results showed that the 0.0625 micromol/L Imatinib or 5 nmol/L Nilotinib alone had no cytotoxic effect on the inhibition of K562/A02 cells. When K562/A02 cells were treated with Imatinib or Nilotinib alone for 48 hours, the expressions of mdr-1 mRNA, der/abl mRNA, P-gp and P210 protein were all down-regulated, furthermore the effect of Nilotinib was stronger than that of Imatinib. The detection of fluorescence intensity revealed that the DNR concentration in K562/A02 cells treated with Imatinib or Nilotinib alone for 48 hours were 7.85% and 12.02% of K562 cells respectively. It is concluded that the tyrosine kinase inhibitors show great effect reversing drug resistance of cells, moreover, the effect of Nilotinib is stronger than that of Imatinib.
本研究旨在探讨酪氨酸激酶抑制剂（TKI）伊马替尼和尼罗替尼对多药耐药细胞系K562/A02的逆转作用。采用逆转录聚合酶链反应（RT-PCR）检测mdr-1 mRNA和bcr-abl mRNA的表达水平。通过蛋白质免疫印迹法检测P-糖蛋白（P-gp）和P210的蛋白水平。采用流式细胞术（FCM）分析柔红霉素（DNR）在K562/A02细胞中的蓄积情况。结果显示，单独使用0.0625 μmol/L伊马替尼或5 nmol/L尼罗替尼对K562/A02细胞无细胞毒性抑制作用。当K562/A02细胞单独用伊马替尼或尼罗替尼处理48小时后，mdr-1 mRNA、der/abl mRNA、P-gp和P210蛋白的表达均下调，且尼罗替尼的作用强于伊马替尼。荧光强度检测显示，单独用伊马替尼或尼罗替尼处理48小时的K562/A02细胞中DNR浓度分别为K562细胞的7.85%和12.02%。结论：酪氨酸激酶抑制剂对细胞耐药逆转作用显著，且尼罗替尼的作用强于伊马替尼。
