论文部分内容阅读
本研究旨在探讨三氧化二砷(ATO)对耐伊马替尼(IM)并有T315I点突变的慢性髓系白血病(CML)细胞株KBM5R的诱导凋亡作用。选择T315I点突变的CML细胞KBM5R和野生型细胞KBM5为研究对象,用MTT法检测KBM5R细胞对IM的耐药性及ATO对KBM5、KBM5R细胞的增殖抑制作用;流式细胞术检测ATO诱导KBM5、KBM5R细胞凋亡;Western blot法检测ATO对KBM5、KBM5R细胞凋亡相关蛋白caspase-3、-8、-9的影响。结果表明,①IM作用于KBM5R细胞的IC50值为12.66±0.565μmol/L,明显高于对KBM5细胞的IC50值(0.303±0.031)μmol/L,两者比较差异具有显著性(p<0.01);②不同浓度ATO作用于KBM5、KBM5R细胞24、48、72、96小时均表现出显著的增殖抑制作用,且具有浓度依赖性和时间依赖性;相同的药物浓度和时间点ATO对KBM5R细胞的增殖抑制作用强于对KBM5细胞;③ATO(2、4、8μmol/L)作用于细胞48小时后KBM5、KBM5R细胞凋亡率均以药物浓度依赖形式增加,且相同药物浓度时KBM5R细胞凋亡率较KBM5细胞高;④4μmol/LATO作用于KBM5、KBM5R细胞24小时后,细胞内cleaved caspase-3、-8、-9蛋白表达明显增加。结论 :KBM5R细胞对IM具有显著的耐药性;ATO对KBM5、KBM5R细胞均具有增殖抑制和诱导凋亡作用,与野生型细胞KBM5比较,ATO对T315I突变的KBM5R细胞的增殖抑制和诱导凋亡作用更强;ATO通过活化凋亡相关蛋白caspase-3、-8、-9诱导细胞KBM5和KBM5R的凋亡。
This study was designed to investigate the effect of arsenic trioxide (ATO) on the apoptosis of chronic myeloid leukemia (CML) cell line KBM5R induced by imatinib (IM) and point mutation T315I. Select the point mutation of T315I CML cells KBM5R and wild-type cells KBM5 as the research object, using MTT assay KBM5R cells resistance to IM and ATO on KBM5, KBM5R cell proliferation inhibition; flow cytometry ATO induced KBM5, KBM5R cells. Western blot was used to detect the effect of ATO on apoptosis-related proteins caspase-3, -8 and -9 in KBM5 and KBM5R cells. The results showed that: (1) The IC50 value ofIM for KBM5R cells was12.66 ± 0.565μmol / L, which was significantly higher than that of KBM5cells (0.303 ± 0.031) μmol / L, the difference was significant (p <0.01); ② At different concentrations of ATO, KBM5 and KBM5R cells showed significant inhibition of proliferation at 24, 48, 72, and 96 hours in a concentration-dependent and time-dependent manner. The proliferation of KBM5R cells induced by ATO at the same drug concentration and time points (2) The apoptotic rates of KBM5 and KBM5R cells increased in a dose-dependent manner after treated with ATO (2, 4, 8μmol / L) for 48 hours, and the apoptotic rates of KBM5R cells in the same drug concentration were higher than those in KBM5R cells KBM5 cells high; ④ 4μmol / LATO role in KBM5, KBM5R cells 24 hours after cleaved caspase-3, -8, -9 protein expression increased significantly. CONCLUSION: KBM5R cells have significant resistance to IM. ATO can inhibit proliferation and induce apoptosis of both KBM5 and KBM5R cells. Compared with wild-type KBM5 cells, ATO can inhibit proliferation and induce apoptosis of T315I mutant KBM5R cells ATO can induce the apoptosis of KBM5 and KBM5R cells by activating caspase-3, -8, -9.