Li Xiao-Feng, Li Jing-He, Wang Chun-Hong, Wang Xiu-Li, Liu Qiu-Ju, Xu Wen, Jia Bo, Qiu Lin, Ma Jun
Department of Oncology & Hematology, Jilin University Second Hospital, Changchun 130041, Jilin Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2011 Jun;19(3):643-7.
This study was aimed to investigate the inducing-apoptosis effect of arsenic trioxide (ATO) on imatinib (IM)-resistant chronic myeloid leukemia (CML) cell line KBM5R with T315I point mutation. CML cell line KBM5R with T315I point mutation and wild-type cell line KBM5 were selected for study. Resistance of KBM5R cells to IM and proliferation of KBM5 and KBM5R cells treated with ATO were detected by MTT; apoptosis of KBM5 and KBM5R cells were quantified by flow cytometry; the expression of apoptosis-related protein caspase-3, -8, -9 was determined by Western blot. The results showed that (1) IC(50) of KBM5R and KBM5 cells treated with IM were 12.66 ± 0.565 µmol/L and 0.303 ± 0.031 µmol/L respectively, and significantly different from each other. (2) the proliferation of KBM5 and KBM5R cells treated with different concentrations of ATO was inhibited in dose- and time-dependent manners at 24, 48, 72, 96 hours, and inhibition of KBM5R cell proliferation was stronger than KBM5 in the same drug concentration and time. (3) the apoptosis rate of KBM5 and KBM5R cells treated with 2, 4, 8 µmol/L ATO for 48 hours increased in a concentration-dependent manner, and the apoptosis rate of KBM5R was higher than that of KBM5 cells in the same drug concentration. (4) the expression of cleaved caspase-3, -8, -9 protein in KBM5 and KBM5R cells treated with 4 µmol/L ATO for 24 hours significantly increased. It is concluded that KBM5R cells are significantly resistant to IM; ATO can inhibit the proliferation and induce the apoptosis of KBM5R and KBM5 cells. As compared with wild-type KBM5 cells, effect of ATO on inhibition of proliferation and induction of apoptosis in KBM5R cells are more stronger. ATO can induce the apoptosis of KBM5 and KBM5R cells through the activation of apoptosis-related caspase-3, -8, -9 protein.
本研究旨在探讨三氧化二砷（ATO）对携带T315I点突变的伊马替尼（IM）耐药慢性髓性白血病（CML）细胞系KBM5R的诱导凋亡作用。选择携带T315I点突变的CML细胞系KBM5R和野生型细胞系KBM5进行研究。采用MTT法检测KBM5R细胞对IM的耐药性以及ATO处理后KBM5和KBM5R细胞的增殖情况；通过流式细胞术定量检测KBM5和KBM5R细胞的凋亡情况；采用蛋白质免疫印迹法检测凋亡相关蛋白caspase-3、-8、-9的表达。结果显示：（1）IM处理后KBM5R和KBM5细胞的IC(50)分别为12.66±0.565µmol/L和0.303±0.031µmol/L，二者差异有统计学意义。（2）在24、48、72、96小时，不同浓度ATO处理KBM5和KBM5R细胞后，细胞增殖呈剂量和时间依赖性抑制，相同药物浓度和时间下，KBM5R细胞增殖抑制作用强于KBM5细胞。（3）2、4、8µmol/L ATO处理KBM5和KBM5R细胞48小时后，细胞凋亡率呈浓度依赖性增加，相同药物浓度下KBM5R细胞凋亡率高于KBM5细胞。（4）4µmol/L ATO处理KBM5和KBM5R细胞24小时后，裂解的caspase-3、-8、-9蛋白表达显著增加。结论：KBM5R细胞对IM显著耐药；ATO可抑制KBM5R和KBM5细胞增殖并诱导其凋亡。与野生型KBM5细胞相比，ATO对KBM5R细胞增殖抑制和凋亡诱导作用更强。ATO可通过激活凋亡相关的caspase-3、-8、-9蛋白诱导KBM5和KBM5R细胞凋亡。
