Inhibition of the ecto-beta subunit of F1F0-ATPase inhibits proliferation and induces apoptosis in acute myeloid leukemia cell lines

Journal of Experimental & Clinical Cancer Research

Pan Jian¹  Ni Jian²  Zhang Min¹  Zhu Xue-Ming¹  Li Yan-Hong¹  Feng Xing¹  Tao Yan-Fang¹  Wang Jian¹  Zhao Wen-Li¹ 

[1] Department of Hematology and Oncology, Children’s Hospital of Soochow University, Suzhou, China;Translational Research Center, Second Hospital, The Second Clinical School, Nanjing Medical University, Nanjing, China

关键词: MV4-11;    HL-60;    Proliferation;    Ecto-ATPase β subunit;    Apoptosis;    Acute myeloid leukemia;   

Others  :  825689
DOI  :  10.1186/1756-9966-31-92

 received in 2012-08-07, accepted in 2012-11-05,  发布年份 2012

【 摘 要 】

Background

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.

Methods

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.

Results

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.

Conclusions

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.

【 授权许可】

   
2012 Wen-Li et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

【 参考文献 】

• [1]Valenti D, Tullo A, Caratozzolo MF, Merafina RS, Scartezzini P, Marra E, Vacca RA: Impairment of F1F0-ATPase, adenine nucleotide translocator and adenylate kinase causes mitochondrial energy deficit in human skin fibroblasts with chromosome 21 trisomy. Biochem J 2010, 431:299-310.
• [2]Percy JM, Pryde JG, Apps DK: Isolation of ATPase I, the proton pump of chromaffin-granule membranes. Biochem J 1985, 231:557-564.
• [3]Zhang X, Gao F, Yu LL, Peng Y, Liu HH, Liu JY, Yin M, Ni J: Dual functions of a monoclonal antibody against cell surface F1F0 ATP synthase on both HUVEC and tumor cells. Acta Pharmacol Sin 2008, 29:942-950.
• [4]Chi SL, Wahl ML, Mowery YM, Shan S, Mukhopadhyay S, Hilderbrand SC, Kenan DJ, Lipes BD, Johnson CE, Marusich MF, et al.: Angiostatin-like activity of a monoclonal antibody to the catalytic subunit of F1F0 ATP synthase. Cancer Res 2007, 67:4716-4724.
• [5]Moser TL, Stack MS, Asplin I, Enghild JJ, Hojrup P, Everitt L, Hubchak S, Schnaper HW, Pizzo SV: Angiostatin binds ATP synthase on the surface of human endothelial cells. Proc Natl Acad Sci U S A 1999, 96:2811-2816.
• [6]Radojkovic C, Genoux A, Pons V, Combes G, de Jonge H, Champagne E, Rolland C, Perret B, Collet X, Terce F, Martinez LO: Stimulation of cell surface F1-ATPase activity by apolipoprotein A-I inhibits endothelial cell apoptosis and promotes proliferation. Arterioscler Thromb Vasc Biol 2009, 29:1125-1130.
• [7]Zick M, Rabl R, Reichert AS: Cristae formation-linking ultrastructure and function of mitochondria. Biochim Biophys Acta 2009, 1793:5-19.
• [8]Plaza H, Whelchel TR, Garczynski SF, Howerth EW, Gherardini FC: Purified outer membranes of Serpulina hyodysenteriae contain cholesterol. J Bacteriol 1997, 179:5414-5421.
• [9]Bienen EJ, Saric M, Pollakis G, Grady RW, Clarkson AB Jr: Mitochondrial development in Trypanosoma brucei brucei transitional bloodstream forms. Mol Biochem Parasitol 1991, 45:185-192.
• [10]Moser TL, Kenan DJ, Ashley TA, Roy JA, Goodman MD, Misra UK, Cheek DJ, Pizzo SV: Endothelial cell surface F1-F0 ATP synthase is active in ATP synthesis and is inhibited by angiostatin. Proc Natl Acad Sci U S A 2001, 98:6656-6661.
• [11]Scotet E, Martinez LO, Grant E, Barbaras R, Jeno P, Guiraud M, Monsarrat B, Saulquin X, Maillet S, Esteve JP, et al.: Tumor recognition following Vgamma9Vdelta2 T cell receptor interactions with a surface F1-ATPase-related structure and apolipoprotein A-I. Immunity 2005, 22:71-80.
• [12]Yonally SK, Capaldi RA: The F(1)F(0) ATP synthase and mitochondrial respiratory chain complexes are present on the plasma membrane of an osteosarcoma cell line: An immunocytochemical study. Mitochondrion 2006, 6:305-314.
• [13]Martinez LO, Jacquet S, Esteve JP, Rolland C, Cabezon E, Champagne E, Pineau T, Georgeaud V, Walker JE, Terce F, et al.: Ectopic beta-chain of ATP synthase is an apolipoprotein A-I receptor in hepatic HDL endocytosis. Nature 2003, 421:75-79.
• [14]Chi SL, Pizzo SV: Angiostatin is directly cytotoxic to tumor cells at low extracellular pH: a mechanism dependent on cell surface-associated ATP synthase. Cancer Res 2006, 66:875-882.
• [15]Das B, Mondragon MO, Sadeghian M, Hatcher VB, Norin AJ: A novel ligand in lymphocyte-mediated cytotoxicity: expression of the beta subunit of H + transporting ATP synthase on the surface of tumor cell lines. J Exp Med 1994, 180:273-281.
• [16]von Haller PD, Donohoe S, Goodlett DR, Aebersold R, Watts JD: Mass spectrometric characterization of proteins extracted from Jurkat T cell detergent-resistant membrane domains. Proteomics 2001, 1:1010-1021.
• [17]Vantourout P, Martinez LO, Fabre A, Collet X, Champagne E: Ecto-F1-ATPase and MHC-class I close association on cell membranes. Mol Immunol 2008, 45:485-492.
• [18]Wang J, Han Y, Liang J, Cheng X, Yan L, Wang Y, Liu J, Luo G, Chen X, Zhao L, et al.: Effect of a novel inhibitory mAb against beta-subunit of F1F0 ATPase on HCC. Cancer Biol Ther 2008, 7:1829-1835.
• [19]Cortes-Hernandez P, Dominguez-Ramirez L, Estrada-Bernal A, Montes-Sanchez DG, Zentella-Dehesa A, de Gomez-Puyou MT, Gomez-Puyou A, Garcia JJ: The inhibitor protein of the F1F0-ATP synthase is associated to the external surface of endothelial cells. Biochem Biophys Res Commun 2005, 330:844-849.
• [20]Wojtczak L: The Crabtree effect: a new look at the old problem. Acta Biochim Pol 1996, 43:361-368.
• [21]Lu ZJ, Song QF, Jiang SS, Song Q, Wang W, Zhang GH, Kan B, Chen LJ, Yang JL, Luo F, et al.: Identification of ATP synthase beta subunit (ATPB) on the cell surface as a non-small cell lung cancer (NSCLC) associated antigen. BMC Cancer 2009, 9:16. BioMed Central Full Text
• [22]Jian P, Li ZW, Fang TY, Jian W, Zhuan Z, Mei LX, Yan WS, Jian N: Retinoic acid induces HL-60 cell differentiation via the upregulation of miR-663. J Hematol Oncol 2011, 4:20. BioMed Central Full Text
• [23]Jian P, Yanfang T, Zhuan Z, Jian W, Xueming Z, Jian N: MMP28 (epilysin) as a novel promoter of invasion and metastasis in gastric cancer. BMC Cancer 2011, 11:200. BioMed Central Full Text
• [24]Pan J, Hu H, Zhou Z, Sun L, Peng L, Yu L, Sun L, Liu J, Yang Z, Ran Y: Tumor-suppressive mir-663 gene induces mitotic catastrophe growth arrest in human gastric cancer cells. Oncol Rep 2010, 24:105-112.
• [25]Ran Y, Pan J, Hu H, Zhou Z, Sun L, Peng L, Yu L, Sun L, Liu J, Yang Z: A novel role for tissue factor pathway inhibitor-2 in the therapy of human esophageal carcinoma. Hum Gene Ther 2009, 20:41-49.
• [26]Mowery YM, Pizzo SV: Targeting cell surface F1F0 ATP synthase in cancer therapy. Cancer Biol Ther 2008, 7:1836-1838.
• [27]Ahmed FE, Wiley JE, Weidner DA, Bonnerup C, Mota H: Surface plasmon resonance (SPR) spectrometry as a tool to analyze nucleic acid-protein interactions in crude cellular extracts. Cancer Genomics Proteomics 2010, 7:303-309.
• [28]Biswas KB, Nabi AH, Arai Y, Nakagawa T, Ebihara A, Ichihara A, Watanabe T, Inagami T, Suzuki F: Aliskiren binds to renin and prorenin bound to (pro)renin receptor in vitro. Hypertens Res 2010, 33:1053-1059.
• [29]Sahlan M, Zako T, Tai PT, Ohtaki A, Noguchi K, Maeda M, Miyatake H, Dohmae N, Yohda M: Thermodynamic characterization of the interaction between prefoldin and group II chaperonin. J Mol Biol 2010, 399:628-636.
• [30]Fukui M, Hinode D, Yokoyama M, Tanabe S, Yoshioka M: Salivary immunoglobulin A directed to oral microbial GroEL in patients with periodontitis and their potential protective role. Oral Microbiol Immunol 2006, 21:289-295.
• [31]Zhou H, Xu Y, Yang Y, Huang A, Wu J, Shi Y: Solution structure of AF-6 PDZ domain and its interaction with the C-terminal peptides from Neurexin and Bcr. J Biol Chem 2005, 280:13841-13847.
• [32]Hu C, Gan N, Chen Y, Bi L, Zhang X, Song L: Detection of microcystins in environmental samples using surface plasmon resonance biosensor. Talanta 2009, 80:407-410.
• [33]Situ C, Wylie AR, Douglas A, Elliott CT: Reduction of severe bovine serum associated matrix effects on carboxymethylated dextran coated biosensor surfaces. Talanta 2008, 76:832-836.
• [34]Rowe PS, Garrett IR, Schwarz PM, Carnes DL, Lafer EM, Mundy GR, Gutierrez GE: Surface plasmon resonance (SPR) confirms that MEPE binds to PHEX via the MEPE-ASARM motif: a model for impaired mineralization in X-linked rickets (HYP). Bone 2005, 36:33-46.
• [35]Aggeler R, Coons J, Taylor SW, Ghosh SS, Garcia JJ, Capaldi RA, Marusich MF: A functionally active human F1F0 ATPase can be purified by immunocapture from heart tissue and fibroblast cell lines. Subunit structure and activity studies. J Biol Chem 2002, 277:33906-33912.
• [36]Grover GJ, Malm J: Pharmacological profile of the selective mitochondrial F1F0 ATP hydrolase inhibitor BMS-199264 in myocardial ischemia. Cardiovasc Ther 2008, 26:287-296.
• [37]Papathanassiu AE, MacDonald NJ, Bencsura A, Vu HA: F1F0-ATP synthase functions as a co-chaperone of Hsp90-substrate protein complexes. Biochem Biophys Res Commun 2006, 345:419-429.
• [38]Reikvam H, Ersvaer E, Bruserud O: Heat shock protein 90 - a potential target in the treatment of human acute myelogenous leukemia. Curr Cancer Drug Targets 2009, 9:761-776.
• [39]Banerji U: Heat shock protein 90 as a drug target: some like it hot. Clin Cancer Res 2009, 15:9-14.