醋酸棉酚诱导淋巴细胞白血病细胞凋亡的作用及联合地塞米松的效应

Effects of Gossypol Acetate on Apoptosis in Primary Cultured Cells from Patients with Lymphoid Leukemia and Its Synergy with Dexamethasone

本研究旨在探讨醋酸棉酚诱导人急、慢性淋巴细胞白血病细胞凋亡的作用及联合地塞米松的效应。用流式细胞仪检测醋酸棉酚对人急、慢性淋巴细胞白血病原代细胞凋亡的影响。用MTT比色法测定醋酸棉酚对人Burkitt淋巴瘤细胞株Raji细胞及正常骨髓单个核细胞存活率的影响。用流式细胞仪检测醋酸棉酚与地塞米松联合诱导Raji细胞的凋亡率。结果表明,≥5μmol/L醋酸棉酚能诱导原代培养的人急性淋巴细胞白血病细胞凋亡,呈浓度及时间依赖性。醋酸棉酚对原代培养的慢性淋巴细胞白血病细胞诱导凋亡的浓度更高,50μmol/L醋酸棉酚作用48 h,急、慢性淋巴细胞白血病细胞的凋亡率分别为(90.4±6.2)%和(51.7±10.3)%。≤10μmol/L醋酸棉酚对正常骨髓单个核细胞的存活率没有明显影响,醋酸棉酚作用48和72 h,对正常骨髓单个核细胞的IC50值分别为Raji细胞的7.1和9.1倍。≥10μmol/L地塞米松能诱导Raji细胞凋亡,不同浓度地塞米松与10μmol/L醋酸棉酚联用,Raji细胞凋亡率明显增加(P<0.05)。结论:醋酸棉酚能诱导原代培养的人急、慢性淋巴细胞白血病细胞凋亡;对正常骨髓单个核细胞的抑制作用明显低于Raji细胞;与地塞米松联用可增强后者诱导Raji细胞凋亡的效应。

To investigate the effects of gossypol acetate on apoptosis in primary cultured cells from patients with acute lymphoblastic leukemia（ALL） and chronic lymphocytic leukemia（CLL） and its synergistic effect with dexamethasone.The apoptosis-inducing effect of gossypol acetate on primary cultured leukemia cells was analyzed by flow cytometry（FCM）.The effect of gossypol acetate on survival rates of Raji cells and mononuclear cells（MNC） from normal bone marrow were evaluated by MTT assay.After co-treatment with gossypol acetate and dexamethasone,the apoptosis rate of Raji cells was detected by FCM.The results showed that gossypol acetate was able to induce apoptosis in primary cultured ALL cells at concentrations of ≥ 5 μmol/L.The effect was concentration and time dependent.Apoptosis-inducing concentration in CLL cells was higher than that in ALL cells.After exposing to 50 μmol/L gossypol acetate for 48 h,the apoptosis rate of ALL and CLL cells were（90.4±6.2）% and（51.7±10.3）% separately.No major growth inhibitory effect was observed in MNC from normal bone marrow when they were exposed to gossypol acetate at concentrations lower than 10 μmol/L.After exposing for 48 and 72 h,the IC50 of gossypol acetate for MNC from normal bone marrow was 7.1 and 9.1 times as much as the IC50 of Raji cells.Co-treatment with 10 μmol/L gossypol acetate and dexamethasone remarkably increased the apoptosis rate of Raji cells.It is concluded that the gossypol acetate has apoptosis-inducing activity in primary cultured leukemia cells from patients diagnosed as ALL and CLL in vitro.The inhibitory effect of gossypol acetate on MNC from normal bone marrow is less prominent than that on Raji cells.Co-treatment with gossypol acetate and dexamethasone notably amplified the pro-apoptosis activity of the latter in Raji cells.