6-异丙基双硫-2'-脱氧鸟苷通过氧化应激诱导骨髓增生异常综合征细胞凋亡
Thiotert Induces Myelodysplastic Syndromes Cells Apoptosis by Activating Oxidative Stress
敬强安 1周超婷 1吴韵怡 2柯霞 2童向民1
作者信息
- 1. 浙江省人民医院(杭州医学院附属人民医院)临床医学研究所,浙江杭州 310006;浙江工业大学生物工程学院,浙江杭州 310014
- 2. 浙江工业大学生物工程学院,浙江杭州 310014
- 折叠
摘要
目的:研究6-异丙基双硫-2'-脱氧鸟苷(thiotert)对骨髓增生异常综合征(MDS)细胞SKM-1增殖及凋亡的影响.方法:采用CCK-8法检测thiotert对SKM-1细胞活力的抑制及GSH、NAC和Z-VAD-FMK对thiotert细胞毒性的逆转作用;EdU细胞增殖检测试剂盒检测细胞增殖情况;流式细胞术检测细胞内的活性氧(ROS)水平;Western blot检测DNA损伤及凋亡相关蛋白的表达.结果:thiotert显著抑制了 SKM-1细胞的增殖能力,诱导SKM-1细胞中ROS水平升高;thiotert处理导致SKM-1细胞内C-PARP、C-Caspase 3和γ-H2AX蛋白表达上调,而BCL-2显著下调;抗氧化剂GSH和NAC及凋亡抑制剂Z-VAD-FMK能部分逆转thiotert对细胞活力的抑制作用.结论:thiotert能够通过介导ROS的产生和促凋亡蛋白的表达来促进MDS细胞发生凋亡.
Abstract
Objective:To explore whether thiotert treatment can inhibit proliferation and induce apoptosis in myelodysplastic syndromes(MDS)cells.Methods:CCK-8 assay was used for determining the cytotoxicity of thiotert to MDS cell line SKM-1 and the reversal effect of GSH,NAC,and Z-VAD-FMK on thiotert-induced inhibition of cell viability.EdU assay was deployed to detect the cell proliferation ability.Intracellular reactive oxygen species(ROS)was measured by flow cytometry after DCFH-DA staining.The expression of DNA damage-and apoptosis-related proteins was detected by Western blot.Results:Thiotert treatment significantly suppressed the cell viability and proliferation ability in SKM-1 cells.A large amount of ROS generation and markedly elevated C-PARP,C-Caspase 3,and γ-H2AX were observed after thiotert administration,while BCL-2 was significantly decreased.In addition,GSH,NAC,and Z-VAD-FMK were able to mitigate the cytotoxicity of thiotert on SKM-1 cells.Conclusion:Thiotert can promote MDS cell apoptosis by mediating ROS production and pro-apoptotic proteins expression.
关键词
骨髓增生异常综合征/6-异丙基双硫-2'-脱氧鸟苷/活性氧/凋亡Key words
myelodysplastic syndromes/thiotert/reactive oxygen species/apoptosis引用本文复制引用
出版年
2024
NETL
NSTL
万方数据