| As2O3诱导NB4、HL-60细胞凋亡的机制研究 |
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| 引用本文： | 张雪莲,周尊海,张利强,王博,贾玉芳,梁爱斌.As2O3诱导NB4、HL-60细胞凋亡的机制研究[J].同济大学学报(医学版),2016,37(3):8-15, 27. |
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| 作者姓名： | 张雪莲 周尊海 张利强 王博 贾玉芳 梁爱斌 |
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| 作者单位： | 同济大学附属杨浦医院内分泌科,上海 200090,同济大学附属杨浦医院内分泌科,上海 200090,上海赛金生物医药有限公司,上海 201203,同济大学附属杨浦医院内分泌科,上海 200090,济宁市第一人民医院妇产科,山东 济宁 272011,同济大学附属同济医院血液科,上海 200065 |
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| 基金项目： | 国家自然科学基金(81270615)；上海市卫生局科研基金(20134470) |
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| 摘 要： | 目的探讨三氧化二砷(As2O3)在体外抑制急性早幼粒白血病(acute promyelocyticleukemia, APL)细胞NB4、HL-60细胞增殖,诱导细胞凋亡的分子机制。方法 通过WST-8法检测上述两个细胞株的增殖抑制曲线;用AnnexinV/PI流式细胞术检测细胞的凋亡;用Real time PCR检测凋亡相关基因Caspase-3、Caspase-8、Caspase-9、Bcl-2、Bax、p53、C-myc、Survivin在As2O3处理前后的表达变化,同时用Westernblot检测凋亡相关蛋白Caspase-3、Caspase-8、Caspase-9、Bcl-2、Bax、P53在As2O3处理前后的表达变化。结果 As2O3能够显著抑制两种细胞增殖, Westernblot检测及Realtime PCR结果显示,As2O3处理引起了两种细胞中Bax、Caspase-3、Caspase-8 、Caspase-9基因表达水平增加,且蛋白表达水平上都出现了活性形式的Caspases。NB4细胞中Survivin、C-myc、Bcl-2的基因表达水平都显著降低,突变型p53蛋白在细胞内的量同样显著下降；HL-60细胞的C-myc表达水平显著降低,但Survivin、Bcl-2表达水平无明显变化。结论 As2O3能在药物临床浓度范围内有效抑制急性早幼粒白血病细胞HL-60、NB4的细胞增殖, 但方式不同；1.5μmol/L浓度的As2O3对NB4细胞生长的抑制体现在诱导细胞分化并诱导细胞凋亡,对HL-60细胞生长的抑制只体现在诱导细胞分化。HL-60细胞中高表达的Survivin、Bcl-2抑制了细胞的凋亡。NB4、HL-60细胞株中P53基因的细胞遗传学变异的不同可能是As2O3对这两种细胞增殖抑制机制差异的主要原因之一。
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| 关 键 词： | As2O3 细胞凋亡 NB4 HL-60 survivin p53 |
| 收稿时间： | 2015/9/12 0:00:00 |
Mechanisms of arsenic trioxide-induced apoptosis in NB4 and HL60 cells |
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| ZHANG Xue-lian,ZHOU Zun-hai,ZHANG Li-qiang,WANG Bo,JIA Yu-fang and LIANG Ai-bin.Mechanisms of arsenic trioxide-induced apoptosis in NB4 and HL60 cells[J].Journal of Tongji University(Medical Science),2016,37(3):8-15, 27. |
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| Authors: | ZHANG Xue-lian ZHOU Zun-hai ZHANG Li-qiang WANG Bo JIA Yu-fang and LIANG Ai-bin |
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| Institution: | Dept.of Endocrinology, Yangpu Hospital, Tongji University, Shanghai 200090, China,Dept.of Endocrinology, Yangpu Hospital, Tongji University, Shanghai 200090, China,Shanghai Cellgene bio Pharmaceutical Co.Ltd., Shanghai 201203, China,Dept.of Endocrinology, Yangpu Hospital, Tongji University, Shanghai 200090, China,Dept.of Gynecology, Jining First People''s Hospital, Jining 272011, Shandong Province, China and Dept.of Hematology, Tongji Hospital, Tongji University, Shanghai 200065, China |
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| Abstract: | Objective To investigate the mechanism of arsenic trioxide (As2O3) -induced apoptosis in acute promyelocytic leukemia(APL)NB4 and HL60 cells.Methods NB4 and HL60 cells were treated with As2O3 at different concentrations. Cell proliferation was determined by WST-8 assay. The apoptosis rate was detected by flow cytometry with annexinV-FITC/PI double staining.The mRNA expression of apoptosis-related genes caspase-3, Caspase-8, Caspase-9, Bcl-2,Bax, the proto-oncogene C-myc, survivin, p53 and granulocytes differentiation antigen CD11b was detected by RT-PCR. The expression of apoptosis-related proteins caspase-3, Caspase-8, Caspase-9, Bcl-2,Bax was determined by Western blotting. Results Arsenic trioxide proliferation of NB4 and HL60 cells with similar sensitivity. As2O3 induced apoptosis of NB4 cells, but induced HL-60 differentiation to mature stage. Western blot detection and RT-PCR results showed that As2O3 treatment caused up-regulation of Bax, caspase-3, Caspase-8, Caspase-9, CD11b mRNA levels, and protein levels of caspase-3, Caspase-8, Caspase-9 in both cell lines. The expression of survivin, C-myc, Bcl-2 and mutated p53 in NB4 cells treated with As2O3 significantly decreased. The expression of C-myc in HL60 cells treated with As2O3 significantly decreased, but survivin and Bcl-2 expression levels were not changed. Conclusion Arsenic trioxide can effectively inhibit proliferation of NB4 and HL60 cells, and induce apoptosis of NB4 cells, while induce HL-60 differentiation to mature stage. These effects may be associated with the altered expression of survivin, Bcl-2 and mutated p53 genes. |
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