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18 pages, 3355 KiB

Open AccessFeature PaperArticle

Orai2 Modulates Store-Operated Ca²⁺ Entry and Cell Cycle Progression in Breast Cancer Cells

by Jose Sanchez-Collado, Jose J. Lopez, Carlos Cantonero, Isaac Jardin, Sergio Regodón, Pedro C. Redondo, Juan Gordillo, Tarik Smani, Gines M. Salido and Juan A. Rosado

Cancers 2022, 14(1), 114; https://doi.org/10.3390/cancers14010114 - 27 Dec 2021

Cited by 17 | Viewed by 3444 | Correction

Abstract

Breast cancer is a heterogeneous disease from the histological and molecular expression point of view, and this heterogeneity determines cancer aggressiveness. Store-operated Ca²⁺ entry (SOCE), a major mechanism for Ca²⁺ entry in non-excitable cells, is significantly remodeled in cancer cells and [...] Read more.

Breast cancer is a heterogeneous disease from the histological and molecular expression point of view, and this heterogeneity determines cancer aggressiveness. Store-operated Ca²⁺ entry (SOCE), a major mechanism for Ca²⁺ entry in non-excitable cells, is significantly remodeled in cancer cells and plays an important role in the development and support of different cancer hallmarks. The store-operated CRAC (Ca²⁺ release-activated Ca²⁺) channels are predominantly comprised of Orai1 but the participation of Orai2 and Orai3 subunits has been reported to modulate the magnitude of Ca²⁺ responses. Here we provide evidence for a heterogeneous expression of Orai2 among different breast cancer cell lines. In the HER2 and triple negative breast cancer cell lines SKBR3 and BT20, respectively, where the expression of Orai2 was greater, Orai2 modulates the magnitude of SOCE and sustain Ca²⁺ oscillations in response to carbachol. Interestingly, in these cells Orai2 modulates the activation of NFAT1 and NFAT4 in response to high and low agonist concentrations. Finally, we have found that, in cells with high Orai2 expression, Orai2 knockdown leads to cell cycle arrest at the G0-G1 phase and decreases apoptosis resistance upon cisplatin treatment. Altogether, these findings indicate that, in breast cancer cells with a high Orai2 expression, Orai2 plays a relevant functional role in agonist-evoked Ca²⁺ signals, cell proliferation and apoptosis resistance. Full article

(This article belongs to the Special Issue Calcium Signaling Remodeling and Functional Role in Cancer Cells)

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32 pages, 7762 KiB

Open AccessArticle

Orai1 Boosts SK3 Channel Activation

by Adéla Tiffner, Valentina Hopl, Romana Schober, Matthias Sallinger, Herwig Grabmayr, Carmen Höglinger, Marc Fahrner, Victoria Lunz, Lena Maltan, Irene Frischauf, Denis Krivic, Rajesh Bhardwaj, Rainer Schindl, Matthias A. Hediger and Isabella Derler

Cancers 2021, 13(24), 6357; https://doi.org/10.3390/cancers13246357 - 17 Dec 2021

Cited by 7 | Viewed by 2957

Abstract

The interplay of SK3, a Ca²⁺ sensitive K⁺ ion channel, with Orai1, a Ca²⁺ ion channel, has been reported to increase cytosolic Ca²⁺ levels, thereby triggering proliferation of breast and colon cancer cells, although a molecular mechanism has remained [...] Read more.

The interplay of SK3, a Ca²⁺ sensitive K⁺ ion channel, with Orai1, a Ca²⁺ ion channel, has been reported to increase cytosolic Ca²⁺ levels, thereby triggering proliferation of breast and colon cancer cells, although a molecular mechanism has remained elusive to date. We show in the current study, via heterologous protein expression, that Orai1 can enhance SK3 K⁺ currents, in addition to constitutively bound calmodulin (CaM). At low cytosolic Ca²⁺ levels that decrease SK3 K⁺ permeation, co-expressed Orai1 potentiates SK3 currents. This positive feedback mechanism of SK3 and Orai1 is enabled by their close co-localization. Remarkably, we discovered that loss of SK3 channel activity due to overexpressed CaM mutants could be restored by Orai1, likely via its interplay with the SK3–CaM binding site. Mapping for interaction sites within Orai1, we identified that the cytosolic strands and pore residues are critical for a functional communication with SK3. Moreover, STIM1 has a bimodal role in SK3–Orai1 regulation. Under physiological ionic conditions, STIM1 is able to impede SK3–Orai1 interplay by significantly decreasing their co-localization. Forced STIM1–Orai1 activity and associated Ca²⁺ influx promote SK3 K⁺ currents. The dynamic regulation of Orai1 to boost endogenous SK3 channels was also determined in the human prostate cancer cell line LNCaP. Full article

(This article belongs to the Special Issue Calcium Signaling Remodeling and Functional Role in Cancer Cells)

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21 pages, 38860 KiB

Open AccessArticle

Orai3 Regulates Pancreatic Cancer Metastasis by Encoding a Functional Store Operated Calcium Entry Channel

by Samriddhi Arora, Jyoti Tanwar, Nutan Sharma, Suman Saurav and Rajender K. Motiani

Cancers 2021, 13(23), 5937; https://doi.org/10.3390/cancers13235937 - 25 Nov 2021

Cited by 7 | Viewed by 2821

Abstract

Store operated Ca²⁺ entry (SOCE) mediated by Orai1/2/3 channels is a highly regulated and ubiquitous Ca²⁺ influx pathway. Although the role of Orai1 channels is well studied, the significance of Orai2/3 channels is still emerging in nature. In this study, we [...] Read more.

Store operated Ca²⁺ entry (SOCE) mediated by Orai1/2/3 channels is a highly regulated and ubiquitous Ca²⁺ influx pathway. Although the role of Orai1 channels is well studied, the significance of Orai2/3 channels is still emerging in nature. In this study, we performed extensive bioinformatic analysis of publicly available datasets and observed that Orai3 expression is inversely associated with the mean survival time of PC patients. Orai3 expression analysis in a battery of PC cell lines corroborated its differential expression profile. We then carried out thorough Ca²⁺ imaging experiments in six PC cell lines and found that Orai3 forms a functional SOCE channel in PC cells. Our in vitro functional assays show that Orai3 regulates PC cell cycle progression, apoptosis and migration. Most importantly, our in vivo xenograft studies demonstrate a critical role of Orai3 in PC tumor growth and secondary metastasis. Mechanistically, Orai3 controls G₁ phase progression, matrix metalloproteinase expression and epithelial-mesenchymal transition in PC cells. Taken together, this study for the first-time reports that Orai3 drives aggressive phenotypes of PC cells, i.e., migration in vitro and metastasis in vivo. Considering that Orai3 overexpression leads to poor prognosis in PC patients, it appears to be a highly attractive therapeutic target. Full article

(This article belongs to the Special Issue Calcium Signaling Remodeling and Functional Role in Cancer Cells)

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16 pages, 8089 KiB

Open AccessArticle

SARAF and EFHB Modulate Store-Operated Ca²⁺ Entry and Are Required for Cell Proliferation, Migration and Viability in Breast Cancer Cells

by Isaac Jardin, Joel Nieto-Felipe, Sandra Alvarado, Raquel Diez-Bello, Jose J. Lopez, Ginés M. Salido, Tarik Smani and Juan A. Rosado

Cancers 2021, 13(16), 4160; https://doi.org/10.3390/cancers13164160 - 19 Aug 2021

Cited by 9 | Viewed by 2286

Abstract

Breast cancer is among the most common malignancies in women. From the molecular point of view, breast cancer can be grouped into different categories, including the luminal (estrogen receptor positive (ER+)) and triple negative subtypes, which show distinctive features and, thus, are sensitive [...] Read more.

Breast cancer is among the most common malignancies in women. From the molecular point of view, breast cancer can be grouped into different categories, including the luminal (estrogen receptor positive (ER+)) and triple negative subtypes, which show distinctive features and, thus, are sensitive to different therapies. Breast cancer cells are strongly dependent on Ca²⁺ influx. Store-operated Ca²⁺ entry (SOCE) has been found to support a variety of cancer hallmarks including cell viability, proliferation, migration, and metastasis. The Ca²⁺ channels of the Orai family and the endoplasmic reticulum Ca²⁺ sensor STIM1 are the essential components of SOCE, but the extent of Ca²⁺ influx is fine-tuned by several regulatory proteins, such as the STIM1 modulators SARAF and EFHB. Here, we show that the expression and/or function of SARAF and EFHB is altered in breast cancer cells and both proteins are required for cell proliferation, migration, and viability. EFHB expression is upregulated in luminal and triple negative breast cancer (TNBC) cells and is essential for full SOCE in these cells. SARAF expression was found to be similar in breast cancer and pre-neoplastic breast epithelial cells, and SARAF knockdown was found to result in enhanced SOCE in pre-neoplastic and TNBC cells. Interestingly, silencing SARAF expression in ER+ MCF7 cells led to attenuation of SOCE, thus suggesting a distinctive role for SARAF in this cell type. Finally, we used a combination of approaches to show that molecular knockdown of SARAF and EFHB significantly attenuates the ability of breast cancer cells to proliferate and migrate, as well as cell viability. In aggregate, SARAF and EFHB are required for the fine modulation of SOCE in breast cancer cells and play an important role in the maintenance of proliferation, migration, and viability in these cells. Full article

(This article belongs to the Special Issue Calcium Signaling Remodeling and Functional Role in Cancer Cells)

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17 pages, 3171 KiB

Open AccessArticle

Store-Operated Calcium Channels Control Proliferation and Self-Renewal of Cancer Stem Cells from Glioblastoma

by Elodie Terrié, Nadine Déliot, Yassine Benzidane, Thomas Harnois, Laëtitia Cousin, Patrick Bois, Lisa Oliver, Patricia Arnault, François Vallette, Bruno Constantin and Valérie Coronas

Cancers 2021, 13(14), 3428; https://doi.org/10.3390/cancers13143428 - 08 Jul 2021

Cited by 10 | Viewed by 2504

Abstract

Glioblastoma is the most frequent and deadly form of primary brain tumors. Despite multimodal treatment, more than 90% of patients experience tumor recurrence. Glioblastoma contains a small population of cells, called glioblastoma stem cells (GSC) that are highly resistant to treatment and endowed [...] Read more.

Glioblastoma is the most frequent and deadly form of primary brain tumors. Despite multimodal treatment, more than 90% of patients experience tumor recurrence. Glioblastoma contains a small population of cells, called glioblastoma stem cells (GSC) that are highly resistant to treatment and endowed with the ability to regenerate the tumor, which accounts for tumor recurrence. Transcriptomic studies disclosed an enrichment of calcium (Ca²⁺) signaling transcripts in GSC. In non-excitable cells, store-operated channels (SOC) represent a major route of Ca²⁺ influx. As SOC regulate the self-renewal of adult neural stem cells that are possible cells of origin of GSC, we analyzed the roles of SOC in cultures of GSC previously derived from five different glioblastoma surgical specimens. Immunoblotting and immunocytochemistry experiments showed that GSC express Orai1 and TRPC1, two core SOC proteins, along with their activator STIM1. Ca²⁺ imaging demonstrated that SOC support Ca²⁺ entries in GSC. Pharmacological inhibition of SOC-dependent Ca²⁺ entries decreased proliferation, impaired self-renewal, and reduced expression of the stem cell marker SOX2 in GSC. Our data showing the ability of SOC inhibitors to impede GSC self-renewal paves the way for a strategy to target the cells considered responsible for conveying resistance to treatment and tumor relapse. Full article

(This article belongs to the Special Issue Calcium Signaling Remodeling and Functional Role in Cancer Cells)

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17 pages, 3859 KiB

Open AccessArticle

The Impact of Extracellular Ca²⁺ and Nanosecond Electric Pulses on Sensitive and Drug-Resistant Human Breast and Colon Cancer Cells

by Julita Kulbacka, Nina Rembiałkowska, Anna Szewczyk, Helena Moreira, Anna Szyjka, Irutė Girkontaitė, Kamil P. Grela and Vitalij Novickij

Cancers 2021, 13(13), 3216; https://doi.org/10.3390/cancers13133216 - 28 Jun 2021

Cited by 13 | Viewed by 2350

Abstract

(1) Background: Calcium electroporation (CaEP) is based on the application of electrical pulses to permeabilize cells (electroporation) and allow cytotoxic doses of calcium to enter the cell. (2) Methods: In this work, we have used doxorubicin-resistant (DX) and non-resistant models of human breast [...] Read more.

(1) Background: Calcium electroporation (CaEP) is based on the application of electrical pulses to permeabilize cells (electroporation) and allow cytotoxic doses of calcium to enter the cell. (2) Methods: In this work, we have used doxorubicin-resistant (DX) and non-resistant models of human breast cancer (MCF-7/DX, MCF-7/WT) and colon cancer cells (LoVo, LoVo/DX), and investigated the susceptibility of the cells to extracellular Ca²⁺ and electric fields in the 20 ns–900 ns pulse duration range. (3) Results: We have observed that colon cancer cells were less susceptible to PEF than breast cancer cells. An extracellular Ca²⁺ (2 mM) with PEF was more disruptive for DX-resistant cells. The expression of glycoprotein P (MDR1, P-gp) as a drug resistance marker was detected by the immunofluorescent (CLSM) method and rhodamine-123 efflux as an MDR1 activity. MDR1 expression was not significantly modified by nanosecond electroporation in multidrug-resistant cells, but a combination with calcium ions significantly inhibited MDR1 activity and cell viability. (4) Conclusions: We believe that PEF with calcium ions can reduce drug resistance by inhibiting drug efflux activity. This phenomenon of MDR mechanism disruption seems promising in anticancer protocols. Full article

(This article belongs to the Special Issue Calcium Signaling Remodeling and Functional Role in Cancer Cells)

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23 pages, 5010 KiB

Open AccessArticle

Orai3-Mediates Cisplatin-Resistance in Non-Small Cell Lung Cancer Cells by Enriching Cancer Stem Cell Population through PI3K/AKT Pathway

by Hiba Abou Daya, Sana Kouba, Hakim Ouled-Haddou, Nazim Benzerdjeb, Marie-Sophie Telliez, Charles Dayen, Henri Sevestre, Loïc Garçon, Frédéric Hague and Halima Ouadid-Ahidouch

Cancers 2021, 13(10), 2314; https://doi.org/10.3390/cancers13102314 - 12 May 2021

Cited by 17 | Viewed by 2444

Abstract

The development of the resistance to platinum salts is a major obstacle in the treatment of non-small cell lung cancer (NSCLC). Among the reasons underlying this resistance is the enrichment of cancer stem cells (CSCs) populations. Several studies have reported the involvement of [...] Read more.

The development of the resistance to platinum salts is a major obstacle in the treatment of non-small cell lung cancer (NSCLC). Among the reasons underlying this resistance is the enrichment of cancer stem cells (CSCs) populations. Several studies have reported the involvement of calcium channels in chemoresistance. The Orai3 channel is overexpressed and constitutes a predictive marker of metastasis in NSCLC tumors. Here, we investigated its role in CSCs populations induced by Cisplatin (CDDP) in two NSCLC cell lines. We found that CDDP treatment increased Orai3 expression, but not Orai1 or STIM1 expression, as well as an enhancement of CSCs markers. Moreover, Orai3 silencing or the reduction of extracellular calcium concentration sensitized the cells to CDDP and led to a reduction in the expression of Nanog and SOX-2. Orai3 contributed to SOCE (Store-operated Calcium entry) in both CDDP-treated and CD133⁺ subpopulation cells that overexpress Nanog and SOX-2. Interestingly, the ectopic overexpression of Orai3, in the two NSCLC cell lines, lead to an increase of SOCE and expression of CSCs markers. Furthermore, CD133⁺ cells were unable to overexpress neither Nanog nor SOX-2 when incubated with PI3K inhibitor. Finally, Orai3 silencing reduced Akt phosphorylation. Our work reveals a link between Orai3, CSCs and resistance to CDDP in NSCLC cells. Full article

(This article belongs to the Special Issue Calcium Signaling Remodeling and Functional Role in Cancer Cells)

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21 pages, 3562 KiB

Open AccessArticle

Furin Prodomain ppFurin Enhances Ca²⁺ Entry Through Orai and TRPC6 Channels’ Activation in Breast Cancer Cells

by Jose J. López, Geraldine Siegfried, Carlos Cantonero, Fabienne Soulet, Jean Descarpentrie, Tarik Smani, Iker Badiola, Simon Pernot, Serge Evrard, Juan A. Rosado and Abdel-Majid Khatib

Cancers 2021, 13(7), 1670; https://doi.org/10.3390/cancers13071670 - 01 Apr 2021

Cited by 8 | Viewed by 2411

Abstract

The intracellular calcium concentration ([Ca²⁺]_i) modulation plays a key role in the regulation of cellular growth and survival in normal cells and failure of [Ca²⁺]_i homeostasis is involved in tumor initiation and progression. Here we showed [...] Read more.

The intracellular calcium concentration ([Ca²⁺]_i) modulation plays a key role in the regulation of cellular growth and survival in normal cells and failure of [Ca²⁺]_i homeostasis is involved in tumor initiation and progression. Here we showed that inhibition of Furin by its naturally occurring inhibitor the prodomain ppFurin in the MDA-MB-231 breast cancer cells resulted in enhanced store-operated calcium entry (SOCE) and reduced the cell malignant phenotype. Expression of ppFurin in a stable manner in MDA-MB-231 and the melanoma MDA-MB-435 cell lines inhibits Furin activity as assessed by in vitro digestion assays. Accordingly, cell transfection experiments revealed that the ppFurin-expressing cells are unable to adequately process the proprotein convertase (PC) substrates vascular endothelial growth factor C (proVEGF-C) and insulin-like growth factor-1 receptor (proIGF-1R). Compared to MDA-MB-435 cells, expression of ppFurin in MDA-MB-231 and BT20 cells significantly enhanced SOCE and induced constitutive Ca²⁺ entry. The enhanced SOCE is impaired by inhibition of Orai channels while the constitutive Ca²⁺ entry is attenuated by silencing or inhibition of TRPC6 or inhibition of Orai channels. Analysis of TRPC6 activation revealed its upregulated tyrosine phosphorylation in ppFurin-expressing MDA-MB-231 cells. In addition, while ppFurin had no effect on MDA-MB-435 cell viability, in MDA-MB-231 cells ppFurin expression reduced their viability and ability to migrate and enhanced their sensitization to the apoptosis inducer hydrogen peroxide and similar results were observed in BT20 cells. These findings suggest that Furin inhibition by ppFurin may be a useful strategy to interfere with Ca²⁺ mobilization, leading to breast cancer cells’ malignant phenotype repression and reduction of their resistance to treatments. Full article

(This article belongs to the Special Issue Calcium Signaling Remodeling and Functional Role in Cancer Cells)

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18 pages, 4910 KiB

Open AccessArticle

Epigenetic Modulation of SPCA2 Reverses Epithelial to Mesenchymal Transition in Breast Cancer Cells

by Monish Ram Makena, Myungjun Ko, Donna Kimberly Dang and Rajini Rao

Cancers 2021, 13(2), 259; https://doi.org/10.3390/cancers13020259 - 12 Jan 2021

Cited by 10 | Viewed by 2658

Abstract

The secretory pathway Ca²⁺-ATPase SPCA2 is a tumor suppressor in triple receptor negative breast cancer (TNBC), a highly aggressive molecular subtype that lacks tailored treatment options. Low expression of SPCA2 in TNBC confers poor survival prognosis in patients. Previous work has [...] Read more.

The secretory pathway Ca²⁺-ATPase SPCA2 is a tumor suppressor in triple receptor negative breast cancer (TNBC), a highly aggressive molecular subtype that lacks tailored treatment options. Low expression of SPCA2 in TNBC confers poor survival prognosis in patients. Previous work has established that re-introducing SPCA2 to TNBC cells restores basal Ca²⁺ signaling, represses mesenchymal gene expression, mitigates tumor migration in vitro and metastasis in vivo. In this study, we examined the effect of histone deacetylase inhibitors (HDACi) in TNBC cell lines. We show that the pan-HDACi vorinostat and the class I HDACi romidepsin induce dose-dependent upregulation of SPCA2 transcript with concurrent downregulation of mesenchymal markers and tumor cell migration characteristic of epithelial phenotype. Silencing SPCA2 abolished the ability of HDACi to reverse epithelial to mesenchymal transition (EMT). Independent of ATPase activity, SPCA2 elevated resting Ca²⁺ levels to activate downstream components of non-canonical Wnt/Ca²⁺ signaling. HDACi treatment led to SPCA2-dependent phosphorylation of CAMKII and β-catenin, turning Wnt signaling off. We conclude that SPCA2 mediates the efficacy of HDACi in reversing EMT in TNBC by a novel mode of non-canonical Wnt/Ca²⁺ signaling. Our findings provide incentive for screening epigenetic modulators that exploit Ca²⁺ signaling pathways to reverse EMT in breast tumors. Full article

(This article belongs to the Special Issue Calcium Signaling Remodeling and Functional Role in Cancer Cells)

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Review

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18 pages, 2220 KiB

Open AccessReview

Ca²⁺ Signaling as the Untact Mode during Signaling in Metastatic Breast Cancer

by Dongun Lee and Jeong Hee Hong

Cancers 2021, 13(6), 1473; https://doi.org/10.3390/cancers13061473 - 23 Mar 2021

Cited by 8 | Viewed by 2974

Abstract

Metastatic features of breast cancer in the brain are considered a common pathology in female patients with late-stage breast cancer. Ca²⁺ signaling and the overexpression pattern of Ca²⁺ channels have been regarded as oncogenic markers of breast cancer. In other words, [...] Read more.

Metastatic features of breast cancer in the brain are considered a common pathology in female patients with late-stage breast cancer. Ca²⁺ signaling and the overexpression pattern of Ca²⁺ channels have been regarded as oncogenic markers of breast cancer. In other words, breast tumor development can be mediated by inhibiting Ca²⁺ channels. Although the therapeutic potential of inhibiting Ca²⁺ channels against breast cancer has been demonstrated, the relationship between breast cancer metastasis and Ca²⁺ channels is not yet understood. Thus, we focused on the metastatic features of breast cancer and summarized the basic mechanisms of Ca²⁺-related proteins and channels during the stages of metastatic breast cancer by evaluating Ca²⁺ signaling. In particular, we highlighted the metastasis of breast tumors to the brain. Thus, modulating Ca²⁺ channels with Ca²⁺ channel inhibitors and combined applications will advance treatment strategies for breast cancer metastasis to the brain. Full article

(This article belongs to the Special Issue Calcium Signaling Remodeling and Functional Role in Cancer Cells)

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2 pages, 502 KiB

Open AccessCorrection

Correction: Sanchez-Collado et al. Orai2 Modulates Store-Operated Ca²⁺ Entry and Cell Cycle Progression in Breast Cancer Cells. Cancers 2021, 14, 114

by Jose Sanchez-Collado, Jose J. Lopez, Carlos Cantonero, Isaac Jardin, Sergio Regodón, Pedro C. Redondo, Juan Gordillo, Tarik Smani, Gines M. Salido and Juan A. Rosado

Cancers 2023, 15(4), 1290; https://doi.org/10.3390/cancers15041290 - 17 Feb 2023

Cited by 1 | Viewed by 706

Abstract

In the original publication [...] Full article

(This article belongs to the Special Issue Calcium Signaling Remodeling and Functional Role in Cancer Cells)

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