Stim and Orai mediate constitutive Ca2+ entry and control endoplasmic reticulum Ca2+ refilling in primary cultures of colorectal carcinoma cells

Estella Zuccolo, Umberto Laforenza, Federica Ferulli, Giorgia Pellavio, Giorgia Scarpellino, Matteo Tanzi, Ilaria Turin, Pawan Faris, Angela Lucariello, Marcello Maestri, Dlzar Ali Kheder, Germano Guerra, Paolo Pedrazzoli, Daniela Montagna, Francesco Moccia

Research output: Contribution to journalArticle

Abstract

Store-operated Ca2+ entry (SOCE) provides a major Ca2+ 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 Ca2+ store. Additionally, Stim and Orai proteins underpin constitutive Ca2+ entry in a growing number of cancer cell types due to the partial depletion of their ER Ca2+ 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 Mn2+-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 Ca2+ influx in pCRC was associated to their lower ER Ca2+ content as compared to mCRC cells. Pharmacological and genetic blockade of Stim1, Stim2, Orai1 and Orai3 prevented ER-dependent Ca2+ release, thereby suggesting that constitutive SOCE maintains ER Ca2+ 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 Ca2+ entry and replenish ER with Ca2+ in primary cultures of CRC cells. However, SOCE is not a promising target to design alternative therapies for CRC.

Original languageEnglish
Pages (from-to)31098-31119
Number of pages22
JournalOncotarget
Volume9
Issue number57
DOIs
Publication statusPublished - Jul 24 2018

Fingerprint

Endoplasmic Reticulum
Colorectal Neoplasms
Pharmacology
Proteins
Neoplasms
Complementary Therapies
Cell Movement
Cell Count
Cell Proliferation
Cell Membrane

Keywords

  • Colorectal cancer
  • Orai
  • Proliferation
  • Stim
  • Store-operated Ca entry

ASJC Scopus subject areas

  • Oncology

Cite this

Stim and Orai mediate constitutive Ca2+ entry and control endoplasmic reticulum Ca2+ 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.

In: Oncotarget, Vol. 9, No. 57, 24.07.2018, p. 31098-31119.

Research output: Contribution to journalArticle

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 Ca2+ entry and control endoplasmic reticulum Ca2+ refilling in primary cultures of colorectal carcinoma cells. In: Oncotarget. 2018 ; Vol. 9, No. 57. pp. 31098-31119.
@article{2ec3363d3de84634ab8805b7507f0056,
title = "Stim and Orai mediate constitutive Ca2+ entry and control endoplasmic reticulum Ca2+ refilling in primary cultures of colorectal carcinoma cells",
abstract = "Store-operated Ca2+ entry (SOCE) provides a major Ca2+ 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 Ca2+ store. Additionally, Stim and Orai proteins underpin constitutive Ca2+ entry in a growing number of cancer cell types due to the partial depletion of their ER Ca2+ 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 Mn2+-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 Ca2+ influx in pCRC was associated to their lower ER Ca2+ content as compared to mCRC cells. Pharmacological and genetic blockade of Stim1, Stim2, Orai1 and Orai3 prevented ER-dependent Ca2+ release, thereby suggesting that constitutive SOCE maintains ER Ca2+ 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 Ca2+ entry and replenish ER with Ca2+ in primary cultures of CRC cells. However, SOCE is not a promising target to design alternative therapies for CRC.",
keywords = "Colorectal cancer, Orai, Proliferation, Stim, Store-operated Ca entry",
author = "Estella Zuccolo and Umberto Laforenza and Federica Ferulli and Giorgia Pellavio and Giorgia Scarpellino and Matteo Tanzi and Ilaria Turin and Pawan Faris and Angela Lucariello and Marcello Maestri and Kheder, {Dlzar Ali} and Germano Guerra and Paolo Pedrazzoli and Daniela Montagna and Francesco Moccia",
year = "2018",
month = "7",
day = "24",
doi = "10.18632/oncotarget.25785",
language = "English",
volume = "9",
pages = "31098--31119",
journal = "Oncotarget",
issn = "1949-2553",
publisher = "Impact Journals LLC",
number = "57",

}

TY - JOUR

T1 - Stim and Orai mediate constitutive Ca2+ entry and control endoplasmic reticulum Ca2+ refilling in primary cultures of colorectal carcinoma cells

AU - Zuccolo, Estella

AU - Laforenza, Umberto

AU - Ferulli, Federica

AU - Pellavio, Giorgia

AU - Scarpellino, Giorgia

AU - Tanzi, Matteo

AU - Turin, Ilaria

AU - Faris, Pawan

AU - Lucariello, Angela

AU - Maestri, Marcello

AU - Kheder, Dlzar Ali

AU - Guerra, Germano

AU - Pedrazzoli, Paolo

AU - Montagna, Daniela

AU - Moccia, Francesco

PY - 2018/7/24

Y1 - 2018/7/24

N2 - Store-operated Ca2+ entry (SOCE) provides a major Ca2+ 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 Ca2+ store. Additionally, Stim and Orai proteins underpin constitutive Ca2+ entry in a growing number of cancer cell types due to the partial depletion of their ER Ca2+ 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 Mn2+-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 Ca2+ influx in pCRC was associated to their lower ER Ca2+ content as compared to mCRC cells. Pharmacological and genetic blockade of Stim1, Stim2, Orai1 and Orai3 prevented ER-dependent Ca2+ release, thereby suggesting that constitutive SOCE maintains ER Ca2+ 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 Ca2+ entry and replenish ER with Ca2+ in primary cultures of CRC cells. However, SOCE is not a promising target to design alternative therapies for CRC.

AB - Store-operated Ca2+ entry (SOCE) provides a major Ca2+ 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 Ca2+ store. Additionally, Stim and Orai proteins underpin constitutive Ca2+ entry in a growing number of cancer cell types due to the partial depletion of their ER Ca2+ 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 Mn2+-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 Ca2+ influx in pCRC was associated to their lower ER Ca2+ content as compared to mCRC cells. Pharmacological and genetic blockade of Stim1, Stim2, Orai1 and Orai3 prevented ER-dependent Ca2+ release, thereby suggesting that constitutive SOCE maintains ER Ca2+ 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 Ca2+ entry and replenish ER with Ca2+ in primary cultures of CRC cells. However, SOCE is not a promising target to design alternative therapies for CRC.

KW - Colorectal cancer

KW - Orai

KW - Proliferation

KW - Stim

KW - Store-operated Ca entry

UR - http://www.scopus.com/inward/record.url?scp=85050510736&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85050510736&partnerID=8YFLogxK

U2 - 10.18632/oncotarget.25785

DO - 10.18632/oncotarget.25785

M3 - Article

AN - SCOPUS:85050510736

VL - 9

SP - 31098

EP - 31119

JO - Oncotarget

JF - Oncotarget

SN - 1949-2553

IS - 57

ER -