Dual functions of a monoclonal antibody against cell surface F1F0 ATP synthase on both HUVEC and tumor cells1
- 1 August 2008
- journal article
- Published by Springer Science and Business Media LLC in Acta Pharmacologica Sinica
- Vol. 29 (8), 942-950
- https://doi.org/10.1111/j.1745-7254.2008.00830.x
Abstract
To generate a monoclonal antibody (McAb) against cell surface F1F0 ATP synthase (ATPase) and observe its antitumoral activity on both human umbilical vein endothelial cells (HUVEC) and tumor cells. Hybridoma cells secreting McAb against ATPase were produced by polyethylene glycol-mediated fusions and screened by ELISA. The specificity of McAb was demonstrated by immunofluorescence and confocal imaging, as well as flow cytometry analysis. After the blockade of surface ATPase with McAb on HUVEC and human breast adenocarcinoma MDA-MB-231 cells, an AT P determination kit and CellTiter96 AQueous Assay (MTS) assay were used to detect the effect of the antibody on extracellular ATP modification and cell proliferation. A cellular cytotoxicity assay in combination with doxorubicin, and a cell migration assay on MDA-MB-231 cells were used to determine the antitumoral activity. Finally, a HUVEC tube formation assay was used to detect the antiangiogenic effect of McAb178-5G10. A monoclonal antibody (McAb178-5G10) against the β-subunit of ATPase was generated, and its reactivity toward HUVEC and tumor cells was studied. We demonstrate that McAb178-5G10 binds to ATPase at the cell surface, where it is able to inhibit AT P synthesis. This antibody also prevents the proliferation of HUVEC and MDA-MB-231 cells. Furthermore, McAb178-5G10 enhances the tumoricidal effects of doxorubicin (PPP<0.01). McAb178-5G10 binds preferentially to cell surface ATPase, blocks ATP synthesis, and exhibits both antiangiogenic and antitumorigenic effects.Keywords
This publication has 22 references indexed in Scilit:
- Angiostatin-Like Activity of a Monoclonal Antibody to the Catalytic Subunit of F1F0 ATP SynthaseCancer Research, 2007
- Proteome analysis of hepatocellular carcinoma cell lines with different metastasis potentials by two-dimensional liquid chromatographyWorld Chinese Journal of Digestology, 2007
- Confocal laser scanning microscopy of whole mouse ovaries: Excellent morphology, apoptosis detection, and spectroscopyCytometry Part A, 2006
- Angiostatin Is Directly Cytotoxic to Tumor Cells at Low Extracellular pH: A Mechanism Dependent on Cell Surface–Associated ATP SynthaseCancer Research, 2006
- Cell surface F1FoATP synthase: A new paradigm?Annals of Medicine, 2006
- Proteomic Analysis of Integral Plasma Membrane ProteinsAnalytical Chemistry, 2004
- Turning the ATP motorNature, 2004
- Endothelial cell surface F 1 -F O ATP synthase is active in ATP synthesis and is inhibited by angiostatinProceedings of the National Academy of Sciences of the United States of America, 2001
- Complementary DNA Cloning and Sequencing of the Chicken Muscle Ecto-ATPaseJournal of Biological Chemistry, 1997
- Continuous cultures of fused cells secreting antibody of predefined specificityNature, 1975