摘要
目的:探讨大剂量维生素C对急性髓系白血病细胞株HL-60、U937及原代急性髓系白血病细胞增殖及凋亡的影响。方法:采用免疫磁珠的方法分选CD34^+细胞,体外培养CD34^+细胞及急性髓系白血病细胞株HL-60、U937。给予不同浓度的维生素C处理各组细胞,采用MTT法检测细胞存活率,Annexin V/PI双染法检测细胞凋亡率,并通过Western blot检测凋亡相关蛋白cleaved caspase-3、cleaved caspase-9及cleaved PARP的表达情况。结果:大剂量维生素C可以抑制HL-60及U937细胞的增殖,且呈浓度依赖性(r=-0.9664;r=-0.9796)。采用不同浓度维生素C(8及20 mmol/L)分别处理HL-60、U937细胞24 h的结果显示,随着维生素C浓度的增加,细胞凋亡率明显增加(r=0.9905;r=0.9971),且凋亡相关蛋白cleaved caspase-3、cleaved caspase-9及cleaved PARP的表达也明显增加。无论有无TET2基因的突变,大剂量维生素C均可抑制原代CD34^+急性髓系白血病细胞的增殖(r=-0.9719;r=-0.9699)、促进其凋亡(r=0.9998;r=0.9901),且呈浓度依赖性。但是却对正常的CD34^+细胞的增殖(r=-0.2032)及凋亡(r=0.1912)无影响。结论:大剂量维生素C能够抑制急性髓系白血病细胞的增殖、促进其凋亡,并且具有选择性杀伤原代CD34^+急性髓系白血病细胞的作用。
Objective:To investigate the effects of high dose vitamin C on proliferation and apoptosis of acute myeloid leukemia(AML)cell lines including HL-60,U937 and primary CD34+leukemia cells in AML.Methods:CD34+cells were sorted by using immunomagnetic cell sorting system,then the primary CD34+leukemia cells,including HL-60 and U937 cell lines were cultured in vitro.Cells in each group were treated with different concentrations of vitamin C,the survival rate of cells was determined by MTT assay,the apoptosis rate of cells was evaluated by Annexin V/PI double staining,the expression of apoptotic proteins-including cleaved caspase 3,cleaved caspase-9 and cleaved PARP were detected by Western blot.Results:The proliferation of HL-60 and U937 cells could be inhibited by high dose vitamin C,which showed a concentration-dependent manner(r=-0.9664;r=-0.9796).HL-60 and U937 cells were treated with different concentrations of vitamin C(8 and 20 mmol/L)for 24 hours,respectively,it was found that with the increasing of vitamin C concentration,cell apoptosis rate was significantly increased(r=0.9905;r=0.9971),and the expression of apoptosis related proteins including cleaved caspase 3,cleaved caspase-9 and cleaved PARP was aslo significantly increased with the increasing of concentration.In addition,it was found that with or without the mutation of TET2,high dose vitamin C could inhibit the proliferation(r=-0.9719;r=-0.9699)and promote the apoptosis(r=0.9998;r=0.9901)of primary CD34^+leukemia cells in AML,which showed a dose-dependent manner,but it showed no effect on the proliferation(r=-0.2032)and apoptosis(r=0.1912)of normal CD34^+cells.Conclusion:High dose vitamin C can inhibit the proliferation and promote the apoptosis of acute myeloid leukemia cells,and selectively kill primary CD34+leukemia cells in AML.
作者
林晓静
邹兴立
赵臻怡
王静
杨竹
倪勋
魏锦
LIN Xiao-Jing;ZOU Xing-Li;ZHAO Zhen-Yi;WANG Jing;YANG Zhu;NI Xun;WEI Jin(Department of Hematology,The Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,Sichuan Province,China)
出处
《中国实验血液学杂志》
CAS
CSCD
北大核心
2020年第3期833-841,共9页
Journal of Experimental Hematology
基金
南充市市校科技战略合作项目(18SXHZ0136)。
