Cargando…
Stim and Orai mediate constitutive Ca(2+) entry and control endoplasmic reticulum Ca(2+) refilling in primary cultures of colorectal carcinoma cells
Store-operated Ca(2+) entry (SOCE) provides a major Ca(2+) entry route in cancer cells. SOCE is mediated by the assembly of Stim and Orai proteins at endoplasmic reticulum (ER)-plasma membrane junctions upon depletion of the ER Ca(2+) store. Additionally, Stim and Orai proteins underpin constitutive...
Autores principales: | , , , , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Impact Journals LLC
2018
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6089563/ https://www.ncbi.nlm.nih.gov/pubmed/30123430 http://dx.doi.org/10.18632/oncotarget.25785 |
_version_ | 1783347043562946560 |
---|---|
author | Zuccolo, Estella Laforenza, Umberto Ferulli, Federica Pellavio, Giorgia Scarpellino, Giorgia Tanzi, Matteo Turin, Ilaria Faris, Pawan Lucariello, Angela Maestri, Marcello Kheder, Dlzar Ali Guerra, Germano Pedrazzoli, Paolo Montagna, Daniela Moccia, Francesco |
author_facet | Zuccolo, Estella Laforenza, Umberto Ferulli, Federica Pellavio, Giorgia Scarpellino, Giorgia Tanzi, Matteo Turin, Ilaria Faris, Pawan Lucariello, Angela Maestri, Marcello Kheder, Dlzar Ali Guerra, Germano Pedrazzoli, Paolo Montagna, Daniela Moccia, Francesco |
author_sort | Zuccolo, Estella |
collection | PubMed |
description | Store-operated Ca(2+) entry (SOCE) provides a major Ca(2+) entry route in cancer cells. SOCE is mediated by the assembly of Stim and Orai proteins at endoplasmic reticulum (ER)-plasma membrane junctions upon depletion of the ER Ca(2+) store. Additionally, Stim and Orai proteins underpin constitutive Ca(2+) entry in a growing number of cancer cell types due to the partial depletion of their ER Ca(2+) reservoir. Herein, we investigated for the first time the structure and function of SOCE in primary cultures of colorectal carcinoma (CRC) established from primary tumor (pCRC) and metastatic lesions (mCRC) of human subjects. Stim1-2 and Orai1-3 transcripts were equally expressed in pCRC and mCRC cells, although Stim1 and Orai3 proteins were up-regulated in mCRC cells. The Mn(2+)-quenching technique revealed that constitutive Ca2(+) entry was significantly enhanced in pCRC cells and was inhibited by the pharmacological and genetic blockade of Stim1, Stim2, Orai1 and Orai3. The larger resting Ca(2+) influx in pCRC was associated to their lower ER Ca(2+) content as compared to mCRC cells. Pharmacological and genetic blockade of Stim1, Stim2, Orai1 and Orai3 prevented ER-dependent Ca(2+) release, thereby suggesting that constitutive SOCE maintains ER Ca(2+) levels. Nevertheless, pharmacological and genetic blockade of Stim1, Stim2, Orai1 and Orai3 did not affect CRC cell proliferation and migration. These data provide the first evidence that Stim and Orai proteins mediate constitutive Ca(2+) entry and replenish ER with Ca(2+) in primary cultures of CRC cells. However, SOCE is not a promising target to design alternative therapies for CRC. |
format | Online Article Text |
id | pubmed-6089563 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Impact Journals LLC |
record_format | MEDLINE/PubMed |
spelling | pubmed-60895632018-08-17 Stim and Orai mediate constitutive Ca(2+) entry and control endoplasmic reticulum Ca(2+) refilling in primary cultures of colorectal carcinoma cells Zuccolo, Estella Laforenza, Umberto Ferulli, Federica Pellavio, Giorgia Scarpellino, Giorgia Tanzi, Matteo Turin, Ilaria Faris, Pawan Lucariello, Angela Maestri, Marcello Kheder, Dlzar Ali Guerra, Germano Pedrazzoli, Paolo Montagna, Daniela Moccia, Francesco Oncotarget Research Paper Store-operated Ca(2+) entry (SOCE) provides a major Ca(2+) entry route in cancer cells. SOCE is mediated by the assembly of Stim and Orai proteins at endoplasmic reticulum (ER)-plasma membrane junctions upon depletion of the ER Ca(2+) store. Additionally, Stim and Orai proteins underpin constitutive Ca(2+) entry in a growing number of cancer cell types due to the partial depletion of their ER Ca(2+) reservoir. Herein, we investigated for the first time the structure and function of SOCE in primary cultures of colorectal carcinoma (CRC) established from primary tumor (pCRC) and metastatic lesions (mCRC) of human subjects. Stim1-2 and Orai1-3 transcripts were equally expressed in pCRC and mCRC cells, although Stim1 and Orai3 proteins were up-regulated in mCRC cells. The Mn(2+)-quenching technique revealed that constitutive Ca2(+) entry was significantly enhanced in pCRC cells and was inhibited by the pharmacological and genetic blockade of Stim1, Stim2, Orai1 and Orai3. The larger resting Ca(2+) influx in pCRC was associated to their lower ER Ca(2+) content as compared to mCRC cells. Pharmacological and genetic blockade of Stim1, Stim2, Orai1 and Orai3 prevented ER-dependent Ca(2+) release, thereby suggesting that constitutive SOCE maintains ER Ca(2+) levels. Nevertheless, pharmacological and genetic blockade of Stim1, Stim2, Orai1 and Orai3 did not affect CRC cell proliferation and migration. These data provide the first evidence that Stim and Orai proteins mediate constitutive Ca(2+) entry and replenish ER with Ca(2+) in primary cultures of CRC cells. However, SOCE is not a promising target to design alternative therapies for CRC. Impact Journals LLC 2018-07-24 /pmc/articles/PMC6089563/ /pubmed/30123430 http://dx.doi.org/10.18632/oncotarget.25785 Text en Copyright: © 2018 Zuccolo et al. http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/) 3.0 (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Paper Zuccolo, Estella Laforenza, Umberto Ferulli, Federica Pellavio, Giorgia Scarpellino, Giorgia Tanzi, Matteo Turin, Ilaria Faris, Pawan Lucariello, Angela Maestri, Marcello Kheder, Dlzar Ali Guerra, Germano Pedrazzoli, Paolo Montagna, Daniela Moccia, Francesco Stim and Orai mediate constitutive Ca(2+) entry and control endoplasmic reticulum Ca(2+) refilling in primary cultures of colorectal carcinoma cells |
title | Stim and Orai mediate constitutive Ca(2+) entry and control endoplasmic reticulum Ca(2+) refilling in primary cultures of colorectal carcinoma cells |
title_full | Stim and Orai mediate constitutive Ca(2+) entry and control endoplasmic reticulum Ca(2+) refilling in primary cultures of colorectal carcinoma cells |
title_fullStr | Stim and Orai mediate constitutive Ca(2+) entry and control endoplasmic reticulum Ca(2+) refilling in primary cultures of colorectal carcinoma cells |
title_full_unstemmed | Stim and Orai mediate constitutive Ca(2+) entry and control endoplasmic reticulum Ca(2+) refilling in primary cultures of colorectal carcinoma cells |
title_short | Stim and Orai mediate constitutive Ca(2+) entry and control endoplasmic reticulum Ca(2+) refilling in primary cultures of colorectal carcinoma cells |
title_sort | stim and orai mediate constitutive ca(2+) entry and control endoplasmic reticulum ca(2+) refilling in primary cultures of colorectal carcinoma cells |
topic | Research Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6089563/ https://www.ncbi.nlm.nih.gov/pubmed/30123430 http://dx.doi.org/10.18632/oncotarget.25785 |
work_keys_str_mv | AT zuccoloestella stimandoraimediateconstitutiveca2entryandcontrolendoplasmicreticulumca2refillinginprimaryculturesofcolorectalcarcinomacells AT laforenzaumberto stimandoraimediateconstitutiveca2entryandcontrolendoplasmicreticulumca2refillinginprimaryculturesofcolorectalcarcinomacells AT ferullifederica stimandoraimediateconstitutiveca2entryandcontrolendoplasmicreticulumca2refillinginprimaryculturesofcolorectalcarcinomacells AT pellaviogiorgia stimandoraimediateconstitutiveca2entryandcontrolendoplasmicreticulumca2refillinginprimaryculturesofcolorectalcarcinomacells AT scarpellinogiorgia stimandoraimediateconstitutiveca2entryandcontrolendoplasmicreticulumca2refillinginprimaryculturesofcolorectalcarcinomacells AT tanzimatteo stimandoraimediateconstitutiveca2entryandcontrolendoplasmicreticulumca2refillinginprimaryculturesofcolorectalcarcinomacells AT turinilaria stimandoraimediateconstitutiveca2entryandcontrolendoplasmicreticulumca2refillinginprimaryculturesofcolorectalcarcinomacells AT farispawan stimandoraimediateconstitutiveca2entryandcontrolendoplasmicreticulumca2refillinginprimaryculturesofcolorectalcarcinomacells AT lucarielloangela stimandoraimediateconstitutiveca2entryandcontrolendoplasmicreticulumca2refillinginprimaryculturesofcolorectalcarcinomacells AT maestrimarcello stimandoraimediateconstitutiveca2entryandcontrolendoplasmicreticulumca2refillinginprimaryculturesofcolorectalcarcinomacells AT khederdlzarali stimandoraimediateconstitutiveca2entryandcontrolendoplasmicreticulumca2refillinginprimaryculturesofcolorectalcarcinomacells AT guerragermano stimandoraimediateconstitutiveca2entryandcontrolendoplasmicreticulumca2refillinginprimaryculturesofcolorectalcarcinomacells AT pedrazzolipaolo stimandoraimediateconstitutiveca2entryandcontrolendoplasmicreticulumca2refillinginprimaryculturesofcolorectalcarcinomacells AT montagnadaniela stimandoraimediateconstitutiveca2entryandcontrolendoplasmicreticulumca2refillinginprimaryculturesofcolorectalcarcinomacells AT mocciafrancesco stimandoraimediateconstitutiveca2entryandcontrolendoplasmicreticulumca2refillinginprimaryculturesofcolorectalcarcinomacells |