Ectopic expression of the ATP synthase β subunit on the membrane of PC-3M cells supports its potential role in prostate cancer metastasis

Li,Wei; Li,Yulin; Li,Gaiyun; Zhou,Zilong; Chang,Xiaona; Xia,Yang; Dong,Xinjie; Liu,Zhijing; Ren,Bo; Liu,Wei; Li,Yilei

doi:10.3892/ijo.2017.3878

Ectopic expression of the ATP synthase β subunit on the membrane of PC-3M cells supports its potential role in prostate cancer metastasis

Authors:
- Wei Li
- Yulin Li
- Gaiyun Li
- Zilong Zhou
- Xiaona Chang
- Yang Xia
- Xinjie Dong
- Zhijing Liu
- Bo Ren
- Wei Liu
- Yilei Li
View Affiliations

Affiliations: The Key Laboratory of Pathobiology, Ministry of Education, The College of Basic Medical Sciences, Jilin University, Changchun, Jilin 130021, P.R. China, Department of Pathology, the Second Clinical Hospital, Jilin University, Changchun, Jilin 130021, P.R. China, Department of Pathology, the First Clinical Hospital, Jilin University, Changchun, Jilin 130021, P.R. China, Department of Pathology, Longgang District Central Hospital of Shenzhen, Shenzhen, Guangdong 518116, P.R. China
Published online on: February 16, 2017 https://doi.org/10.3892/ijo.2017.3878
Pages: 1312-1320

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Metastatic prostate cancer is associated with high mortality rates. Identification of metastasis-related proteins may facilitate the development of novel therapies for the treatment of metastatic disease. In the present study, we aimed to identify prostate cancer metastasis-associated membrane proteins. We developed a phage-displayed 7-mer peptide library to screen the target peptides that were specifically bound to PC-3M cells with subtractive panning from normal prostate cells and PC-3 prostate cancer cells. A novel short peptide (B04) was found to have high affinity to highly metastatic PC-3M cells. ATP synthase β subunit (ATP5B) was then identified as a binding partner of B04 on the PC-3M cell surface. ATP5B was expressed on the PC-3M cell membrane and on highly malignant human prostate cancer specimens, as shown using multiple methodologies. Furthermore, ATP5B-positive gold particles were detected on the cellular and mitochondrial membranes by immunoelectromicroscopy. These results implied the possibility that ATP5B may translocate from the inner mitochondrial membrane to the outer surface of PC-3M cells. Additional analysis showed that incubation of B04 with PC-3M cells reduced the detection of ATP5B by western blotting and flow cytometry and significantly inhibited the proliferation, invasion and metastasis of PC-3M cells. In conclusion, ATP5B, as a binding partner of a metastasis-related short peptide (B04) on prostate cancer cells, is involved in promoting prostate cancer metastasis. In conclusion, ATP5B may be a promising biomarker and therapeutic target for highly metastatic malignancies.

View Figures

View References

1	Jemal A, Siegel R, Xu J and Ward E: Cancer statistics, 2010. CA Cancer J Clin. 60:277–300. 2010. View Article : Google Scholar : PubMed/NCBI
2	Hanahan D and Weinberg RA: Hallmarks of cancer: the next generation. Cell. 144:646–674. 2011. View Article : Google Scholar : PubMed/NCBI
3	Kauffman EC, Robinson VL, Stadler WM, Sokoloff MH and Rinker-Schaeffer CW: Metastasis suppression: The evolving role of metastasis suppressor genes for regulating cancer cell growth at the secondary site. J Urol. 169:1122–1133. 2003. View Article : Google Scholar : PubMed/NCBI
4	Hanash SM, Madoz-Gurpide J and Misek DE: Identification of novel targets for cancer therapy using expression proteomics. Leukemia. 16:478–485. 2002. View Article : Google Scholar : PubMed/NCBI
5	Gao J, Gao Y, Ju Y, Yang J, Wu Q, Zhang J, Du X, Wang Z, Song Y, Li H, et al: Proteomics-based generation and characterization of monoclonal antibodies against human liver mitochondrial proteins. Proteomics. 6:427–437. 2006. View Article : Google Scholar
6	Liu B, Huang L, Sihlbom C, Burlingame A and Marks JD: Towards proteome-wide production of monoclonal antibody by phage display. J Mol Biol. 315:1063–1073. 2002. View Article : Google Scholar : PubMed/NCBI
7	Chi SL and Pizzo SV: Cell surface F1Fo ATP synthase: A new paradigm? Ann Med. 38:429–438. 2006. View Article : Google Scholar : PubMed/NCBI
8	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 cellsurface as a non-small cell lung cancer (NSCLC) associated antigen. BMC Cancer. 9:162009. View Article : Google Scholar
9	Dowling P, Meleady P, Dowd A, Henry M, Glynn S and Clynes M: Proteomic analysis of isolated membrane fractions from superin- vasivecancer cells. Biochim Biophys Acta. 1774:93–101. 2007. View Article : Google Scholar
10	Kim BW, Choo HJ, Lee JW, Kim JH and Ko YG: Extracellular ATP isgenerated by ATP synthase complex in adipocyte lipid rafts. Exp Mol Med. 36:476–485. 2004. View Article : Google Scholar : PubMed/NCBI
11	Hong D, Jaron D, Buerk DG and Barbee KA: Transport- dependent calcium signaling in spatially segregated cellular caveolar domains. Am J Physiol Cell Physiol. 294:C856–C866. 2008. View Article : Google Scholar
12	Bae TJ, Kim MS, Kim JW, Kim BW, Choo HJ, Lee JW, Kim KB, Lee CS, Kim JH, Chang SY, et al: Lipid raft proteome reveals ATP synthase complex in the cell surface. Proteomics. 4:3536–3548. 2004. View Article : Google Scholar : PubMed/NCBI
13	Ribatti D: Chicken chorioallantoic membrane angiogenesis model. Methods Mol Biol. 843:47–57. 2012. View Article : Google Scholar : PubMed/NCBI
14	Kariya Y, Kato K, Hayashizaki Y, Himeno S, Tarui S and Matsubura K: Revision of consensus sequence of human Alu repeats a review. Gene. 53:1–10. 1987. View Article : Google Scholar
15	Boyer PD: The ATP synthase - a splendid molecular machine. Annu Rev Biochem. 66:717–749. 1997. View Article : Google Scholar
16	Larimer BM, Thomas WD, Smith GP and Deutscher SL: Affinity maturation of an ERBB2-targeted SPECT imaging peptide by in vivo phage display. Mol Imaging Biol. 16:449e582014. View Article : Google Scholar
17	Goetz M and Wang TD: Molecular imaging in gastrointestinal endoscopy. Gastroenterology. 138:828e33.e8212010. View Article : Google Scholar
18	Senior AE: ATP synthesis by oxidative phosphorylation. Physiol Rev. 68:177–231. 1988.PubMed/NCBI
19	Stock D, Leslie AG and Walker JE: Molecular architecture of the rotary motor in ATP synthase. Science. 286:1700–1705. 1999. View Article : Google Scholar : PubMed/NCBI
20	Ma Z, Cao M, Liu Y, He Y, Wang Y, Yang C, Wang W, Du Y, Zhou M and Gao F: Mitochondrial F1Fo-ATP synthase translocates to cell surface in hepatocytes and has high activity in tumor-like acidic and hypoxic environment. Acta Biochim Biophys Sin (Shanghai). 42:530–537. 2010. View Article : Google Scholar
21	Champagne E, Martinez LO and Collet Xand Barbaras R: Ecto-F1Fo ATP synthase/F1 ATPase: metabolic and immunological functions. Curr Opin Lipidol. 17:279–284. 2006. View Article : Google Scholar : PubMed/NCBI
22	Jung KH, Song SH, Paik JY, Koh BH, Choe YS, Lee EJ, Kim BT and Lee KH: Direct targeting of tumor cell F₁F₀ ATP-synthase by radioiodine angiostatin in vitro and in vivo. Cancer Biother Radiopharm. 22:704–712. 2007. View Article : Google Scholar : PubMed/NCBI
23	Das B, Mondragon MO, Sadeghian M, Hatcher VB and 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. 180:273–281. 1994. View Article : Google Scholar : PubMed/NCBI
24	Arakaki N, Nagao T, Niki R, Toyofuku A, Tanaka H, Kuramoto Y, Emoto Y, Shibata H, Magota K and Higuti T: Possible role of cell surface H -ATP synthase in the extracellular ATP synthesis and proliferation of human umbilical vein endothelial cells. Mol Cancer Res. 1:931–939. 2003.PubMed/NCBI
25	Moser TL, Stack MS, Asplin I, Enghild JJ, Højrup P, Everitt L, Hubchak S, Schnaper HW and Pizzo SV: Angiostatin binds ATP synthase on the surface of human endothelial cells. Proc Natl Acad Sci USA. 96:2811–2816. 1999. View Article : Google Scholar : PubMed/NCBI
26	Moser TL, Kenan DJ, Ashley TA, Roy JA, Goodman MD, Misra UK, Cheek DJ and Pizzo SV: Endothelial cell surface F1-F0 ATP synthase is active in ATP synthesis and is inhibited by angiostatin. Proc Natl Acad Sci USA. 98:6656–6661. 2001. View Article : Google Scholar : PubMed/NCBI
27	Notari L, Arakaki N, Mueller D, Meier S, Amaral J and Becerra SP: Pigment epithelium-derived factor binds to cell- surface F₁-ATP synthase. FEBS J. 277:2192–2205. 2010. View Article : Google Scholar : PubMed/NCBI
28	Rumjahn SM, Yokdang N, Baldwin KA, Thai J and Buxton IL: Purinergic regulation of vascular endothelial growth factor signaling in angiogenesis. Br J Cancer. 100:1465–1470. 2009. View Article : Google Scholar : PubMed/NCBI
29	Chi SL and Pizzo SV: Angiostatin is directly cytotoxic to tumor cells at low extracellular pH: A mechanism dependent on cell surface-associated ATP synthase. Cancer Res. 66:875–882. 2006. View Article : Google Scholar : PubMed/NCBI
30	Schwiebert EM and Zsembery A: Extracellular ATP as a signaling molecule for epithelial cells. Biochim Biophys Acta. 1615:7–32. 2003. View Article : Google Scholar : PubMed/NCBI
31	Bodin P and Burnstock G: Increased release of ATP from endothelial cells during acute inflammation. Inflamm Res. 47:351–354. 1998. View Article : Google Scholar : PubMed/NCBI
32	Di Virgilio F: Dr. Jekyll/Mr. Hyde: The dual role of extracellular ATP. J Auton Nerv Syst. 81:59–63. 2000. View Article : Google Scholar : PubMed/NCBI
33	Ralevic V and Burnstock G: Receptors for purines and pyrimidines. Pharmacol Rev. 50:413–492. 1998.PubMed/NCBI
34	Wen LT and Knowles AF: Extracellular ATP and adenosine induce cell apoptosis of human hepatoma Li-7A cells via the A3 adenosine receptor. Br J Pharmacol. 140:1009–1018. 2003. View Article : Google Scholar : PubMed/NCBI
35	Schwiebert LM, Rice WC, Kudlow BA, Taylor AL and Schwiebert EM: Extracellular ATP signaling and P2X nucleotide receptors in monolayers of primary human vascular endothelial cells. Am J Physiol Cell Physiol. 282:C289–C301. 2002. View Article : Google Scholar : PubMed/NCBI
36	Burnstock G: Purinergic signaling and vascular cell proliferation and death. Arterioscler Thromb Vasc Biol. 22:364–373. 2002. View Article : Google Scholar : PubMed/NCBI