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目的研究细胞分化帮助剂(CDA-Ⅱ)促进低浓度三氧化二砷(As_2O_3)对肝癌细胞株HepG-2,BEL-7402凋亡的协同效应。方法用四甲基偶氮唑蓝(MTT)法检测单用 CDA-Ⅱ或As_2O_3和联用对肝癌细胞株 HepG-2,BEL-7402生长及抑制率的影响:用乳酸脱氧酶(LDH)法检测药物对细胞膜的毒性反应;流式细胞仪(FCM)分析细胞周期和凋亡率变化。结果单用 CDA-Ⅱ和As_2O_3分别为3.0mg/ml 和50μmol/L 时,肝癌细胞株 BEL-7402,HepG-2发生凋亡达到最大值,而联用5μmol/L As_2O_3+1mg/ml CDA-Ⅱ可达到同样效果;结果显示:该浓度下 HepG_2的抑制率可达88%;BEL-7402的抑制率在100%;LDH 检测表明联用比单用 As_2O_310μmol/L 更安全。流式细胞仪结果显示凋亡率联合组为40.2%,明显高于单用组 As_2O_3(11.3%)或 CDA-Ⅱ(8.7%)。结论细胞分化帮助剂1mg/ml CDA-Ⅱ联用5μmol/L As_2O_3可促进肝癌细胞株 HepG-2,BEL-7402凋亡,可以明显降低 As_2O_3的使用浓度,减少对正常细胞的毒副作用。二者联用具有协同作用。
Objective To study the synergistic effect of CDA-Ⅱ on apoptosis of HepG-2 and BEL-7402 cells induced by low concentration of arsenic trioxide (As 2 O 3). Methods The effects of combination of CDA-Ⅱ or As_2O_3 alone and on HepG-2 and BEL-7402 hepatocellular carcinoma cell lines were detected by MTT assay. The effect of lactate dehydrogenase (LDH) Toxicity of the drug on the cell membrane was detected. The changes of cell cycle and apoptosis rate were analyzed by flow cytometry (FCM). Results The apoptosis rates of BEL-7402 and HepG-2 cells reached the maximum when CDA-Ⅱ and As_2O_3 were 3.0mg / ml and 50μmol / L, respectively. However, the combination of 5μmol / L As_2O_3 + 1mg / Ⅱ can achieve the same effect; the results showed that: the concentration of HepG_2 inhibition rate up to 88%; BEL-7402 inhibition rate of 100%; LDH test showed that combined with As_2O_310μmol / L safer. The results of flow cytometry showed that the apoptosis rate in combination group was 40.2%, which was significantly higher than that in single group (11.3%) or CDA-Ⅱ (8.7%). Conclusions Apoptosis of hepatocellular carcinoma cell lines HepG-2 and BEL-7402 can be significantly inhibited by 1mg / ml CDA-Ⅱ combined with 5μmol / L As_2O_3, which can significantly reduce the concentration of As_2O_3 and decrease the toxicity to normal cells. The combination of the two has a synergistic effect.