维生素C联合地西他滨对人急性T淋巴细胞白血病细胞株Jurkat细胞凋亡及周期相关基因的影响

Effect of vitamin C combined with decitabine on cell apoptosis and cycle-related genes of human acute T lymphoblastic leukemia cell line Jurkat

目的探讨维生素C联合地西他滨对人急性T淋巴细胞白血病细胞株Jurkat细胞凋亡及周期相关基因的影响。方法采用维生素C 100μg/mL(维生素C组)、地西他滨10μmol/L(地西他滨组)及维生素C 100μg/mL+地西他滨10μmol/L(联合组)处理Jurkat细胞;对照组未加药物。培养24及48h后采用倒置显微镜观察各组细胞的生长状态,培养48h后采用实时荧光定量PCR检测细胞凋亡及周期相关基因的表达。结果培养48h,维生素C组细胞数量较对照组减少(P<0.01),细胞周期相关基因细胞周期依赖性激酶4及细胞周期蛋白E1(cyclin E1)表达下调(P<0.05)。地西他滨组细胞数量较对照组和维生素C组减少(P<0.05),细胞凋亡相关基因Bcl-2拮抗/杀伤因子(Bak)、BH3相互作用域死亡激动剂(Bid)、凋亡相关因子(Fas)、凋亡相关因子配体(FasL)及细胞周期相关基因cyclin E1、p21表达较对照组和维生素C组上调,增殖细胞核抗原(PCNA)、Bax表达较对照组上调(P<0.05)。联合组细胞数量较对照组和维生素C组减少(P<0.05),Bcl-2相关死亡启动因子、Bak、Bid、Fas、cyclin E1、p21表达较对照组和维生素C组上调,PCNA、Bax表达较对照组上调,FasL表达较其他三组上调(P<0.05)。结论维生素C联合地西他滨能进一步上调Jurkat细胞凋亡相关基因及细胞周期抑制基因p21,起到协同抗肿瘤作用。

Objective To investigate the effect of vitamin C combined with decitabine on the cell apoptosis and cycle-related genes of human acute T lymphoblastic leukemia cell line Jurkat.Methods The Jurkat cells were treated with vitamin C 100 μg/mL(group A),decitabine 10 μmol/L(group B) and vitamin C 100 μg/mL + decitabine 10 μmol/L(group C).The control group(group D) was not given any drugs.The cell growth status was observed by inverted microscope 24 and 48 hours after cell culture.The expressions of cell apoptosis and cycle-related genes were detected 48 hours after cell culture.Results After cell culture for 48 hours, the cell number of group A was less than that of group D(P<0.01).The expressions of cyclin-dependent kinase 4 and cyclin E1 were downregulated in group A than those in group D(P<0.05).The cell number of group B was less than that of groups of D and A(P<0.05).The expressions of Bcl-2 antagonist/killer 1(Bak),BH3 interacting domain death agonist(Bid),factor-related apoptosis(Fas),factor-related apoptosis ligand(FasL),cyclin E1 and cell cycle suppressor gene p21 were higher in group B than those in groups of A and D,and those of Bcl-2-assaciated X(Bax) and proliferating cell nuclear antigen(PCNA) were higher in group B than those in groups of D and A(P<0.05).The cell number of group C was less than that of groups of D and A(P<0.05).The expressions of Bcl-2-associated death promoter, Bak, Bid, Fas, cyclin E1 and p21 were higher in group C than those in groups of A and D,those of Bax and PCNA were higher in group C than those in group D,and that of FasL was higher in group C than that in groups of A,B and D(P<0.05).Conclusion Vitamin combined with decitabine can further upregulate Jurkat cell apoptosis gene and cell cycle suppressor gene p21,and plays a synergistic anti-tumor effect.