Cui Ting-Yun, Chen Bao-An, Ding Jia-Hua, Gao Chong, Cheng Jian, Bao Wen, Zhong Yue-Jiao, Shan Xue-Yun, Gao Feng, Xia Guo-Hua, Schmitt Anita, Schmitt Michael
Department of Hematolagy, Zhongda Hospital, Southeast University Medical College, Nanjing 210009, Jiangsu Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2011 Feb;19(1):28-33.
This study was aimed to investigate the relevance of nilotinib in combination with tetrandrine (Tet) on reversing multidrug resistance and inducing apoptosis of K562/A02 cell line and its mechanism. Methyl-thiazol tetrazolium (MTT) assay was employed to examine the pharmacological effect of nilotinib or Tet alone on K562/A02 cell line, the IC(50) of daunorubicin (DNR) on K562/A02 cell line treated with nilotinib and Tet was calculated; the flow cytometry (FCM) was employed to detect the apoptosis rate of K562/A02. The expression of bax/survivin mRNA was determined by RT-PCR, and the expression of bax/survivin protein was assayed by Western blot. The results showed that after being treated by 5 nmol/L nilotinib or 1.0 µml/L Tet for 48 hours, IC(50) of DNR to K562/A02 was 5.71 ± 0.72 mg/L or 6.52 ± 0.43 mg/L, respectively, while in their combined treatment, IC(50) decreased to 3.12 ± 0.13 mg/L. Nilotinib or Tet alone could increase DNR-inducing apoptosis rate of K562/A02 cell, while the apoptosis rate of K562/A02 increased remarkably in combination treatment of nilotinib with Tet. After being treated with 5 nmol/L nilotinib or 1.0 µml/L Tet alone for 48 hours, the expressions of bax mRNA and BAX protein was up-regulated, while both effects were more obvious in combination treatment of nilotinib with Tet. Treatment with 5 nmol/L nilotinib or 1.0 µmol/L Tet alone for 48 hours down-regulated the expression of survivin mRNA and its protein, while treatment of nilotinib in combination with Tet had more significant effect on down-regulation of their expression. It is concluded that the K562/A02 cells are resistant to DNR, nilotinib or Tet alone both can partially reverse resistance of K562/A02 cells to DNR, increase the apoptosis rate of K562/A02 cells. Combination of nilotinib with Tet shows obvious synergistic action, mechanism of which may associate with up-regulation of bax mRNA and BAX protein expressions and down-regulation of survivin mRNA and its protein expressions.
本研究旨在探讨尼罗替尼联合粉防己碱（Tet）对K562/A02细胞系多药耐药逆转及诱导凋亡的相关性及其机制。采用甲基噻唑基四氮唑蓝（MTT）法检测尼罗替尼或粉防己碱单独作用于K562/A02细胞系的药理作用，计算柔红霉素（DNR）对尼罗替尼和粉防己碱处理后的K562/A02细胞系的半数抑制浓度（IC50）；采用流式细胞术（FCM）检测K562/A02细胞的凋亡率。通过逆转录聚合酶链反应（RT-PCR）检测bax/survivin mRNA的表达，采用蛋白质免疫印迹法检测bax/survivin蛋白的表达。结果显示，5 nmol/L尼罗替尼或浓度为1.0 μmol/L粉防己碱作用48小时后，DNR对K562/A02细胞系的IC50分别为5.71±0.72 mg/L或6.52±0.43 mg/L，而联合用药时，IC50降至3.12±0.13 mg/L。尼罗替尼或粉防己碱单独作用均可增加DNR诱导的K562/A02细胞凋亡率，而尼罗替尼与粉防己碱联合处理时，K562/A02细胞凋亡率显著增加。5 nmol/L尼罗替尼或1.0 μmol/L粉防己碱单独作用48小时后，bax mRNA和BAX蛋白表达上调，而尼罗替尼与粉防己碱联合处理时两种作用更明显。5 nmol/L尼罗替尼或1.0 μmol/L粉防己碱单独作用48小时可下调survivin mRNA及其蛋白表达，而尼罗替尼与粉防己碱联合处理对其表达下调作用更显著。结论：K562/A02细胞对DNR耐药，尼罗替尼或粉防己碱单独作用均可部分逆转K562/A02细胞对DNR的耐药性，增加K562/A02细胞凋亡率。尼罗替尼与粉防己碱联合显示出明显的协同作用，其机制可能与上调bax mRNA和BAX蛋白表达及下调survivin mRNA及其蛋白表达有关。
