Differential expression of PMCA2 mRNA isoforms in a cohort of Spanish patients with breast tumor types

Romero‑Lorca,Alicia; Gaibar,Maria; Armesilla,Angel  Luis; Fernandez‑Santander,Ana; Novillo,Apolonia

doi:10.3892/ol.2018.9540

Differential expression of PMCA2 mRNA isoforms in a cohort of Spanish patients with breast tumor types

Authors:
- Alicia Romero‑Lorca
- Maria Gaibar
- Angel Luis Armesilla
- Ana Fernandez‑Santander
- Apolonia Novillo
View Affiliations

Affiliations: Department of Basic Biomedical Sciences, Faculty of Biomedical and Health Sciences, Universidad Europea de Madrid, Villaviciosa de Odón, Madrid 28670, Spain, Faculty of Science and Engineering, School of Pharmacy, University of Wolverhampton, Wolverhampton, West Midlands WV1 1LY, UK
Published online on: October 2, 2018 https://doi.org/10.3892/ol.2018.9540
Pages: 6950-6959
Copyright: © Romero‑Lorca et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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

The present study examined the mRNA expression levels of different isoforms of the plasma membrane calcium ATPase 2 (PMCA2) gene generated by alternative splicing at the first intracellular loop (site A) and C‑terminal region (site C) in 85 human breast cancer tumor and 69 adjacent non‑tumor tissues. Associations were identified between the expression of PMCA2 splice isoforms and the following clinical variables: Estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2) status, tumor size, staging and histological classification, and lymph node status. Transcripts including splice site A or splice site C were amplified by reverse transcription‑quantitative polymerase chain reaction using PMCA2 isoform‑specific primers. Tumor and adjacent tissues were determined to express the different PMCA2 splice isoforms 2w, 2x and 2z (site A), and 2b (site C). The mRNA levels for these variants indicated high biological variability, but increased expression was observed in breast tumor tissues, compared with in adjacent tissues. Significantly increased PMCA2x/b expression levels were detected in breast tumor tissues histologically classified as lobulillar, compared with in ductal‑types breast tumor tissues (P<0.028). Furthermore, PMCA2z expression was significantly associated with PR status (P<0.024, compared with in PR‑negative tumor tissues), and PMCA2w expression was significantly associated with ER status (P<0.048, increased in ER‑positive tumor tissues, compared with ER‑negative tumor tissues). Finally, PMCA2b was overexpressed in HER2‑positive tumor tissues, compared with in HER2‑negative tumor tissues (P<0.014). The data demonstrated the differential mRNA expression of a number of splice site A and C variants of PMCA2 in breast tumor and adjacent tissues, depending on tumor hormone receptor status and histological classification. In agreement with previous data, PMCA2b was overexpressed in HER2‑positive tumor tissues, indicating that high mRNA levels of this variant could be a marker of poor prognosis.

View Figures

View References

1	Strehler EE, Filoteo AG, Penniston JT and Caride AJ: Plasma-membrane Ca(2+) pumps: Structural diversity as the basis for functional versatility. Biochem Soc Trans. 35:919–922. 2007. View Article : Google Scholar : PubMed/NCBI
2	Padányi R, Pászty K, Hegedűs L, Varga K, Papp B, Penniston JT and Enyedi Á: Multifaceted plasma membrane Ca(2+) pumps: From structure to intracellular Ca(2+) handling and cancer. Biochim Biophys Acta. 1863:1351–1363. 2016. View Article : Google Scholar : PubMed/NCBI
3	Stafford N, Wilson C, Oceandy D, Neyses L and Cartwright EJ: The plasma membrane calcium ATPases and their role as Major new players in human disease. Physiol Rev. 97:1089–1125. 2017. View Article : Google Scholar : PubMed/NCBI
4	Brini M, Calì T, Ottolini D and Carafoli E: The plasma membrane calcium pump in health and disease. FEBS J. 280:5385–5397. 2013. View Article : Google Scholar : PubMed/NCBI
5	Bruce JIE: Metabolic regulation of the PMCA: Role in cell death and survival. Cell Calcium. 69:28–36. 2018. View Article : Google Scholar : PubMed/NCBI
6	Váradi A, Molnár E and Ashcroft SJ: A unique combination of plasma membrane Ca2+-ATPase isoforms is expressed in islets of Langerhans and pancreatic beta-cell lines. Biochem J. 314:663–669. 1996. View Article : Google Scholar : PubMed/NCBI
7	Strehler EE: Plasma membrane calcium ATPases: From generic Ca(2+) sump pumps to versatile systems for fine-tuning cellular Ca(2.). Biochem Biophys Res Commun. 460:26–33. 2015. View Article : Google Scholar : PubMed/NCBI
8	Talarico EF Jr, Kennedy BG, Marfurt CF, Loeffler KU and Mangini NJ: Expression and immunolocalization of plasma membrane calcium ATPase isoforms in human corneal epithelium. Mol Vis. 11:169–178. 2005.PubMed/NCBI
9	Talarico EF Jr and Mangini NJ: Alternative splice variants of plasma membrane calcium-ATPases in human corneal epithelium. Exp Eye Res. 85:869–879. 2007. View Article : Google Scholar : PubMed/NCBI
10	Calì T, Brini M and Carafoli E: Regulation of cell calcium and role of plasma membrane calcium ATPases. Int Rev Cell Mol Biol. 332:259–296. 2017. View Article : Google Scholar : PubMed/NCBI
11	Lee WJ, Roberts-Thomson SJ, Holman NA, May FJ, Lehrbach GM and Monteith GR: Expression of plasma membrane calcium pump isoform mRNAs in breast cancer cell lines. Cell Signal. 14:1015–1022. 2002. View Article : Google Scholar : PubMed/NCBI
12	Lee WJ, Roberts-Thomson SJ and Monteith GR: Plasma membrane calcium-ATPase 2 and 4 in human breast cancer cell lines. Biochem Biophys Res Commun. 337:779–783. 2005. View Article : Google Scholar : PubMed/NCBI
13	Farmer P, Bonnefoi H, Becette V, Tubiana-Hulin M, Fumoleau P, Larsimont D, Macgrogan G, Bergh J, Cameron D, Goldstein D, et al: Identification of molecular apocrine breast tumours by microarray analysis. Oncogene. 24:4660–4671. 2005. View Article : Google Scholar : PubMed/NCBI
14	VanHouten J, Sullivan C, Bazinet C, Ryoo T, Camp R, Rimm DL, Chung G and Wysolmerski J: PMCA2 regulates apoptosis during mammary gland involution and predicts outcome in breast cancer. Proc Natl Acad Sci USA. 107:11405–11410. 2010. View Article : Google Scholar : PubMed/NCBI
15	Jeong J, VanHouten JN, Dann P, Kim W, Sullivan C, Yu H, Liotta L, Espina V, Stern DF, Friedman PA and Wysolmerski JJ: PMCA2 regulates HER2 protein kinase localization and signaling and promotes HER2-mediated breast cancer. Proc Natl Acad Sci USA. 113:E282–E290. 2016. View Article : Google Scholar : PubMed/NCBI
16	Heim R, Hug M, Iwata T, Strehler EE and Carafoli E: Microdiversity of human-plasma-membrane calcium-pump isoform 2 generated by alternative RNA splicing in the N-terminal coding region. Eur J Biochem. 205:333–340. 1992. View Article : Google Scholar : PubMed/NCBI
17	Hilfiker H, Guerini D and Carafoli E: Cloning and expression of isoform 2 of the human plasma membrane Ca2+ ATPase. Functional properties of the enzyme and its splicing products. J Biol Chem. 269:26178–26183. 1994.PubMed/NCBI
18	Stauffer TP, Hilfiker H, Carafoli E and Strehler EE: Quantitative analysis of alternative splicing options of human plasma membrane calcium pump genes. J Biol Chem. 268:25993–26003. 1993.PubMed/NCBI
19	Chicka MC and Strehler EE: Alternative splicing of the first intracellular loop of plasma membrane Ca2+-ATPase isoform 2 alters its membrane targeting. J Biol Chem. 278:18464–18470. 2003. View Article : Google Scholar : PubMed/NCBI
20	Strehler EE and Zacharias DA: Role of alternative splicing in generating isoform diversity among plasma membrane calcium pumps. Physiol Rev. 81:21–50. 2001. View Article : Google Scholar : PubMed/NCBI
21	Edge SB, Byrd DR, Compton CC, Fritz AG, Greene FL and Trotti A: AJCC (American Joint Committee on Cancer) Cancer Staging Manual. Springer (eds); 7th. New York: 2010
22	Zerbino DR, Achuthan P, Akanni W, Amode MR, Barrell D, Bhai J, Billis K, Cummins C, Gall A, Girón CG, et al: Ensembl 2018. Nucleic Acids Res. 46:D754–D761. 2018. View Article : Google Scholar : PubMed/NCBI
23	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
24	Monteith GR, Davis FM and Roberts-Thomson SJ: Calcium channels and pumps in cancer: Changes and consequences. J Biol Chem. 287:31666–31673. 2012. View Article : Google Scholar : PubMed/NCBI
25	Roberts-Thomson SJ, Curry MC and Monteith GR: Plasma membrane calcium pumps and their emerging roles in cancer. World J Biol Chem. 1:248–253. 2010. View Article : Google Scholar : PubMed/NCBI
26	Mu L, Tuck D, Katsaros D, Lu L, Schulz V, Perincheri S, Menato G, Scarampi L, Harris L and Yu H: Favorable outcome associated with an IGF-1 ligand signature in breast cancer. Breast Cancer Res Treat. 133:321–331. 2012. View Article : Google Scholar : PubMed/NCBI
27	Curry MC, Luk NA, Kenny PA, Roberts-Thomson SJ and Monteith GR: Distinct regulation of cytoplasmic calcium signals and cell death pathways by different plasma membrane calcium ATPase isoforms in MDA-MB-231 breast cancer cells. J Biol Chem. 287:28598–28608. 2012. View Article : Google Scholar : PubMed/NCBI
28	Curry M, Roberts-Thomson SJ and Monteith GR: PMCA2 silencing potentiates MDA-MB-231 breast cancer cell death initiated with the Bcl-2 inhibitor ABT-263. Biochem Biophys Res Commun. 478:1792–1797. 2016. View Article : Google Scholar : PubMed/NCBI
29	Krebs J: The plethora of PMCA isoforms: Alternative splicing and differential expression. Biochim Biophys Acta. 1853:2018–2024. 2015. View Article : Google Scholar : PubMed/NCBI
30	Lopreiato R, Giacomello M and Carafoli E: The plasma membrane calcium pump: New ways to look at an old enzyme. J Biol Chem. 289:10261–10268. 2014. View Article : Google Scholar : PubMed/NCBI
31	Brini M, Di Leva F, Ortega CK, Domi T, Ottolini D, Leonardi E, Tosatto SC and Carafoli E: Deletions and mutations in the acidic lipid-binding region of the plasma membrane Ca2+ pump: A study on different splicing variants of isoform 2. J Biol Chem. 285:30779–30791. 2010. View Article : Google Scholar : PubMed/NCBI
32	Reinhardt TA and Horst RL: Ca2+-ATPases and their expression in the mammary gland of pregnant and lactating rats. Am J Physiol. 276:C796–C802. 1999. View Article : Google Scholar : PubMed/NCBI
33	Brini M and Carafoli E: Calcium pumps in Health and disease. Physiol Rev. 89:1341–1378. 2009. View Article : Google Scholar : PubMed/NCBI
34	Reinhardt TA, Lippolis JD, Shull GE and Horst RL: Null mutation in the gene encoding plasma membrane Ca2+-ATPase isoform 2 impairs calcium transport into milk. J Biol Chem. 279:42369–42373. 2004. View Article : Google Scholar : PubMed/NCBI
35	Reinhardt TA and Lippolis JD: Mammary gland involution is associated with rapid down regulation of major mammary Ca2+-ATPases. Biochem Biophys Res Commun. 378:99–102. 2009. View Article : Google Scholar : PubMed/NCBI
36	Silverstein RS and Tempel BL: Atp2b2, encoding plasma membrane Ca2+-ATPase type 2, (PMCA2) exhibits tissue-specific first exon usage in hair cells, neurons, and mammary glands of mice. Neuroscience. 141:245–257. 2006. View Article : Google Scholar : PubMed/NCBI
37	Faddy HM, Smart CE, Xu R, Lee GY, Kenny PA, Feng M, Rao R, Brown MA, Bissell MJ, Roberts-Thomson SJ and Monteith GR: Localization of plasma membrane and secretory calcium pumps in the mammary gland. Biochem Biophys Res Commun. 369:977–981. 2008. View Article : Google Scholar : PubMed/NCBI
38	Hill JK, Williams DE, LeMasurier M, Dumont RA, Strehler EE and Gillespie PG: Splice-site A choice targets plasma-membrane Ca2+-ATPase isoform 2 to hair bundles. J Neurosci. 26:6172–6180. 2006. View Article : Google Scholar : PubMed/NCBI
39	Antalffy G, Caride AJ, Pászty K, Hegedus L, Padanyi R, Strehler EE and Enyedi A: Apical localization of PMCA2w/b is enhanced in terminally polarized MDCK cells. Biochem Biophys Res Commun. 410:322–327. 2011. View Article : Google Scholar : PubMed/NCBI
40	Padányi R, Xiong Y, Antalffy G, Lór K, Pászty K, Strehler EE and Enyedi A: Apical scaffolding protein NHERF2 modulates the localization of alternatively spliced plasma membrane Ca2+ pump 2B variants in polarized epithelial cells. J Biol Chem. 285:31704–31712. 2010. View Article : Google Scholar : PubMed/NCBI
41	Peters AA, Milevskiy MJ, Lee WC, Curry MC, Smart CE, Saunus JM, Reid L, da Silva L, Marcial DL, Dray E, et al: The calcium pump plasma membrane Ca(2+)-ATPase 2 (PMCA2) regulates breast cancer cell proliferation and sensitivity to doxorubicin. Sci Rep. 6:255052016. View Article : Google Scholar : PubMed/NCBI
42	Desmedt C, Zoppoli G, Sotiriou C and Salgado R: Transcriptomic and genomic features of invasive lobular breast cancer. Semin Cancer Biol. 44:98–105. 2017. View Article : Google Scholar : PubMed/NCBI
43	Holton M, Yang D, Wang W, Mohamed TM, Neyses L and Armesilla AL: The interaction between endogenous calcineurin and the plasma membrane calcium-dependent ATPase is isoform specific in breast cancer cells. FEBS Lett. 581:4115–4119. 2007. View Article : Google Scholar : PubMed/NCBI
44	Kosiorek M, Podszywalow-Bartnicka P, Zylinska L and Pikula S: NFAT1 and NFAT3 cooperate with HDAC4 during regulation of alternative splicing of PMCA isoforms in PC12 cells. PLoS One. 9:e991182014. View Article : Google Scholar : PubMed/NCBI
45	Eroles P, Bosch A, Pérez-Fidalgo JA and Lluch A: Molecular biology in breast cancer: Intrinsic subtypes and signaling pathways. Cancer Treat Rev. 38:698–707. 2012. View Article : Google Scholar : PubMed/NCBI