目的:观察地西他滨对TP53突变弥漫大B细胞淋巴瘤(diffuse large B cell lymphoma,DLBCL)细胞株DB增殖、凋亡及周期的影响,并评估地西他滨联合治疗TP53突变DLBCL患者的疗效及安全性.方法:体外培养细胞,不同浓度(5～80 μmol/L)地西他滨和阿霉素作用于DB细胞后采用CCK-8法计算增殖抑制率;地西他滨(20 μmol/L)和阿霉素(12 μmol/L)作用于DB细胞24/48 h,采用Annexin V-FITC/PI及流式细胞术检测凋亡率及细胞周期;比较不同组间细胞增殖、凋亡及周期的差异.另收集5例地西他滨联合治疗TP53突变DLBCL患者的临床资料进行回顾性总结.结果:对DB细胞的增殖抑制作用随着地西他滨浓度的增加(5、10、20、40、80 μmol/L)而增强,联合阿霉素(12 μmol/L)后抑制率较单药明显增大[(78.51±1.19)％vs(40.80± 1.62)％,(87.48±0.29)％vs(46.83±1.47)％,(92.59±0.15)％vs(47.07±1.50)％,(94.57±0.58)％vs(52.68±0.84)％,(95.56±0.53)％vs(54.95±1.07)％,P＜0.01];采用 IC50 浓度(20 μmol/L 地西他滨、12 μmol/L阿霉素)药物处理细胞,抑制率随时间增加(12、24、48 h)而增大[(50.40±0.19)％、(61.65±0.16)％、(85.08±0.78)％,P＜0.01].与空白对照组比较,地西他滨作用于DB细胞24 h显示S期细胞比例增加[(10.54±0.40)％vs(2.03±0.01)％,P＜0.01],48 h 显示 G2/M 期细胞比例增加[(17.45±0.19)％vs(3.69±0.09)％,P＜0.01].5例复发/难治TP53突变DLBCL患者使用含有地西他滨方案治疗后,总有效率为40％(2/5),主要不良反应为骨髓抑制,未出现治疗相关死亡.结论:地西他滨可增强阿霉素对TP53突变DLBCL细胞株的增殖抑制作用,阻滞细胞周期.含地西他滨方案治疗TP53突变DLBCL显示出一定疗效,安全性良好,值得进一步探索.
Effect of decitabine combined with adriamycin on TP53 mutated diffuse large B lymphoma cell line and its clinical significance
Objective:To observe the effect of decitabine(DAC)on the proliferation,apoptosis and cell cycle of TP53 mutated diffuse large B cell lymphoma(DLBCL)cell line DB,and evaluate the efficacy and safety of DAC combined therapy in patients with TP53 mutant DLBCL.Methods:Cells were cultured in vitro.The different concentrations(5-80 μmol/L)of DAC and adriamycin(ADM)act on DB cells and the CCK-8 method was used to calculate the proliferation inhibition rate.DAC(20 μmol/L)and ADM(12 μmol/L)act on DB cells 24,48 h,An-nexin V-FITC/PI and flow cytometry were used to detect the apoptosis rate and cell cycle.The differences in cell proliferation,apoptosis and cell cycle in different groups were compared.Clinical data of 5 patients with TP53 mutated DLBCL treated with DAC combination was retrospectively collected and summarized.Results:The inhib-itory effect on the proliferation of DB cells increased with the increase of DAC concentration(5,10,20,40,80 μmol/L),and the inhibition rate in combination with ADM(12 μmol/L)was significantly higher than that of single drug([78.51±1.19]％vs[40.80±1.62]％,[87.48±0.29]％vs[46.83±1.47]％,[92.59±0.15]％vs[47.07±1.50]％,[94.57±0.58]％vs[52.68±0.84]％.[95.56±0.53]％vs[54.95±1.07]％,P＜0.01).When the cells were treated with IC50(20 μmol/L DAC,12 μmol/L ADM),the inhibition rate increased([50.40±0.19]％,[61.65±0.16]％,[85.08±0.78]％)with the increase of time(12,24,48 h)(P＜0.01).Compared with the blank control group,the percentage of DB cells in S phase increased after treatment with DAC for 24 h([10.54±0.40]％vs[2.03±0.01]％,P＜0.01),and the proportion of cells in G2/M phase increased at 48 h([17.45±0.19]％vs[3.69±0.09]％,P＜0.01).The five patients with recurrent refractory TP53 mu-tation DLBCL treated with DAC regimen,the total effective rate was 40％(2/5).The main adverse event was myelosuppression and there was no treatment-related death.Conclusion:DAC can enhance the inhibitory effect of ADM on the proliferation of TP53 mutant DLBCL cell line,and block cell cycle.The regimen containing DAC shows a certain efficacy and safety in the treatment of TP53 mutation DLBCL,which is worthy of further exploration.
diffuse large B cell lymphomarelapsed or refractoryTP53 mutationadriamycindecitabine
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