Inhibition of the ecto-beta subunit of F1F0-ATPase inhibits proliferation and induces apoptosis in acute myeloid leukemia cell lines.
Inhibition of the ecto-beta subunit of F1F0-ATPase inhibits proliferation and induces apoptosis in acute myeloid leukemia cell lines.
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DOI：
10.1186/1756-9966-31-92
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发表时间：
2012-11-09
期刊：
影响因子：
--
通讯作者：
Jian P
中科院分区：
文献类型：
--
作者：
Wen-Li Z;Jian W;Yan-Fang T;Xing F;Yan-Hong L;Xue-Ming Z;Min Z;Jian N;Jian P
白血病是一种造血祖细胞的异质性克隆性疾病，是世界性的健康问题，尤其是在儿童时期。F1F0 ATPase是线粒体内的一种酶，表达于肿瘤细胞的质膜上，其抑制作用可诱导抗血管生成和抗肿瘤活性。用聚乙二醇介导法制备抗ATPase的单抗，并用ELISA法进行筛选。用McAb阻断细胞表面的ATPase，分析细胞的增殖、细胞周期和细胞凋亡情况。我们在包括急性髓系白血病在内的11个白血病细胞系中检测到F1F0ATPaseβ亚单位在细胞膜上的表达。我们制备的McAb7E10能识别天然和重组的ATPase亚基，其解离常数(Kd)为3.26E-10。我们证明McAb7E10能与细胞表面的ATPase结合，从而抑制β的合成。McAb7E10对体外培养的急性髓系白血病细胞有明显的增殖抑制作用：50 μg/mLMcAb7E10对MV4-11和HL-60细胞的相对抑制率分别为69.6%和81.9%。细胞周期分析显示McAb7E10能显著诱导MV4-11和HL-60细胞的凋亡：5、10和50ug/mLMcAb7E10处理的MV4-11细胞的相对凋亡率分别为3.6 ± 0.83%、8.4 ± 1.69%和17.3 ± 2.56%，而对照组为1.5% ± 0.85%(p <； 0.01)。5、10和50ug/mLMcAb7E10处理的HL-60细胞的相对凋亡率分别为5.5 ± 2.37%、11.3 ± 3.62%和19.9 ± 3.31%，而对照组为1.56% ± 0.97%(p <； 0.01)。Annexin V染色显示，50 μg/mLMcAb7E10处理的MV4-11和HL-60细胞的相对凋亡率分别为50.5% ± 7.04%和32.9% ± 4.52%，显著高于免疫球蛋白对照组的21.9% ± 3.11%和15.3% ± 3.95%，p 和lt； 0.01。提示ATPaseβ亚基的异位表达可能是恶性血液病的肿瘤相关抗原。F1F0ATPaseβ亚单位为急性髓系白血病和血液系统恶性肿瘤的免疫治疗提供了一个潜在的靶点。
Leukemia, a heterogeneous clonal disorder of hematopoietic progenitor cells, presents a world-wide health problem, especially in childhood. F1F0 ATPase, an inner mitochondrial enzyme, is expressed on the plasma membrane of tumor cells, and its inhibition induces both anti-angiogenic and anti-tumorigenic activity. Monoclonal Antibody (McAb) against ATPase was produced by polyethylene glycol-mediated fusions and screened by ELISA. Proliferation, cell cycle and apoptosis of cells were analyzed when the surface ATPase of cells was blockaded with McAb. We detected cell-membrane expression of the F1F0 ATPase β subunit on 0.1% to 56% of the 11 cell lines derived from leukemia, including acute myeloid leukemia (AML). We produced a monoclonal antibody, McAb7E10, which recognizes both the native and recombinant ATPase β subunit, with a dissociation constant (KD) of 3.26E–10. We demonstrate that McAb7E10 binds to ATPase at the cell surface, where it is able to inhibit ATP synthesis. McAb7E10 significantly inhibited proliferation of AML cell lines in vitro: the relative inhibitory rates of 50 μg/mL McAb7E10 treated MV4-11and HL-60 cells were 69.6% and 81.9% respectively. Cell cycle analysis indicated that McAb7E10 significantly induced apoptosis in MV4-11 and HL-60 cells: the relative rates of apoptosis in 5, 10 and 50ug/mL McAb7E10 treated MV4-11 cells was 3.6 ± 0.83%, 8.4 ± 1.69% and 17.3 ± 2.56% compared to 1.5% ± 0.85% in mouse IgG treated cells (p < 0.01). The relative rate of apoptosis in 5, 10 and 50ug/mL McAb7E10 treated HL-60 cells was 5.5 ± 2.37%, 11.3 ± 3.62% and 19.9 ± 3.31% compared to 1.56% ± 0.97% in mouse IgG treated cells (p < 0.01). Annexin V staining demonstrated that the relative apoptotic rates in 50 μg/mL McAb7E10 treated MV4-11 and HL-60 cells were 50.5% ± 7.04% and 32.9% ± 4.52%, respectively, significantly higher than IgG control antibody treated cells were 21.9% ± 3.11% and 15.3% ± 3.95%, p < 0.01. These findings indicate that ectopic expression of ATPase β subunit may be a tumor-associated antigen in hematological malignancies. The F1F0 ATPase β subunit provides a potential target for immunotherapy in AML and hematological malignancies.
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影响因子：
5.6
作者：
Sahlan, Muhamad;Zako, Tamotsu;Yohda, Masafumi
通讯作者：
Yohda, Masafumi
视黄酸通过上调 miR-663 诱导 HL-60 细胞分化
影响因子：
28.5
作者：
Jian, Pan;Li, Zhao Wen;Jian, Ni
通讯作者：
Jian, Ni
影响因子：
--
作者：
Fukui, M.;Hinode, D.;Yoshioka, M.
通讯作者：
Yoshioka, M.
DOI：
10.1073/pnas.96.6.2811
发表时间：
1999-03-16
影响因子：
11.1
作者：
Moser, TL;Stack, MS;Pizzo, SV
通讯作者：
Pizzo, SV
影响因子：
4.2
作者：
Pan, Jian;Hu, Hai;Ran, Yuliang
通讯作者：
Ran, Yuliang