Multiple Roles of Protein Kinase A in Arachidonic Acid–Mediated Ca2+ Entry and Tumor-Derived Human Endothelial Cell Migration

Fiorio, Alessandra; Genova, Tullio; Pupo, Emanuela; Tomatis, Cristiana; Genazzani, Armando A.; Zaninetti, Roberta; Munaron, Luca

doi:10.1158/1541-7786.mcr-10-0002

Cited by 36 publications

(33 citation statements)

References 41 publications

(75 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For instance, TECs from human RCC over-express genes that promote endothelial proliferation, survival, motility, and cell adhesion [79]. In agreement with our results, TECs harvested from breast cancer generate abnormal Ca 2+ signals when exposed to arachidonic acid (AA), a pro-angiogenic intracellular messenger [81]–[82]. Normal ECs may undergo genetic and epigenetic modifications by receiving DNA directly from the tumor through apoptotic bodies, or following mRNA and microRNA transfer by microvesicles [80].…”

Section: Discussionsupporting

confidence: 80%

Store-Operated Ca2+ Entry Is Remodelled and Controls In Vitro Angiogenesis in Endothelial Progenitor Cells Isolated from Tumoral Patients

et al. 2012

View full text Add to dashboard Cite

BackgroundEndothelial progenitor cells (EPCs) may be recruited from bone marrow to sustain tumor vascularisation and promote the metastatic switch. Understanding the molecular mechanisms driving EPC proliferation and tubulogenesis could outline novel targets for alternative anti-angiogenic treatments. Store-operated Ca2+ entry (SOCE), which is activated by a depletion of the intracellular Ca2+ pool, regulates the growth of human EPCs, where is mediated by the interaction between the endoplasmic reticulum Ca2+-sensor, Stim1, and the plasmalemmal Ca2+ channel, Orai1. As oncogenesis may be associated to the capability of tumor cells to grow independently on Ca2+ influx, it is important to assess whether SOCE regulates EPC-dependent angiogenesis also in tumor patients.Methodology/Principal FindingsThe present study employed Ca2+ imaging, recombinant sub-membranal and mitochondrial aequorin, real-time polymerase chain reaction, gene silencing techniques and western blot analysis to investigate the expression and the role of SOCE in EPCs isolated from peripheral blood of patients affected by renal cellular carcinoma (RCC; RCC-EPCs) as compared to control EPCs (N-EPCs). SOCE, activated by either pharmacological (i.e. cyclopiazonic acid) or physiological (i.e. ATP) stimulation, was significantly higher in RCC-EPCs and was selectively sensitive to BTP-2, and to the trivalent cations, La3+ and Gd3+. Furthermore, 2-APB enhanced thapsigargin-evoked SOCE at low concentrations, whereas higher doses caused SOCE inhibition. Conversely, the anti-angiogenic drug, carboxyamidotriazole (CAI), blocked both SOCE and the intracellular Ca2+ release. SOCE was associated to the over-expression of Orai1, Stim1, and transient receptor potential channel 1 (TRPC1) at both mRNA and protein level The intracellular Ca2+ buffer, BAPTA, BTP-2, and CAI inhibited RCC-EPC proliferation and tubulogenesis. The genetic suppression of Stim1, Orai1, and TRPC1 blocked CPA-evoked SOCE in RCC-EPCs.ConclusionsSOCE is remodelled in EPCs from RCC patients and stands out as a novel molecular target to interfere with RCC vascularisation due to its ability to control proliferation and tubulogenesis.

show abstract

Section: Discussionsupporting

confidence: 80%

Store-Operated Ca2+ Entry Is Remodelled and Controls In Vitro Angiogenesis in Endothelial Progenitor Cells Isolated from Tumoral Patients

et al. 2012

View full text Add to dashboard Cite

show abstract

“…Interestingly, BTECs showed lower TRPM8 mRNA and protein levels than normal ECs (Fig. 1 Aii), further confirming the different expression pattern of Ca 2+ -related proteins in BTECs compared with normal ECs (Fiorio Pla et al, 2008, 2010, 2012b; Pupo et al, 2011).…”

Section: Resultssupporting

confidence: 59%

“…Cells were grown on glass gelatin-coated coverslips (gelatin coating was avoided for experiments using Tyrode physiological solution without Ca 2+ ) at a density of 5,000 cells/cm 2 for 24 to 48 h. 2 to 5 h before experiments, cells were starved in DMEM with 0% FCS. Cells were next loaded (45 min at 37°C) with 2 µM Fura-2 AM (Invitrogen) for ratiometric cytosolic Ca 2+ ([Ca 2+ ]i) measurements as previously described (Fiorio Pla et al, 2010, 2012b). During the experiments, cells were continuously bathed with a microperfusion system.…”

Section: Methodsmentioning

confidence: 99%

TRPM8 inhibits endothelial cell migration via a non-channel function by trapping the small GTPase Rap1

Genova

Grolez

Camillo

et al. 2017

Journal of Cell Biology

Self Cite

View full text Add to dashboard Cite

show abstract

“…We have previously shown that AA-induced Ca 2+ entry promotes the organization of BTEC into vessel-like structures in vitro , a phenomenon that is partly dependent on cell migration (Fiorio Pla et al , 2008). HMVEC migration, on the other hand, was not effected by AA (Fiorio Pla et al , 2010). Thus, wound-healing assays were conducted with BTEC to further confirm the role of AA on EC migration.…”

Section: Resultsmentioning

confidence: 90%

TRPV4 mediates tumor-derived endothelial cell migration via arachidonic acid-activated actin remodeling

et al. 2011

View full text Add to dashboard Cite

Changes in intracellular calcium [Ca2+]i levels control critical cytosolic and nuclear events that are involved in the initiation and progression of tumor angiogenesis in endothelial cells (ECs). Therefore, the mechanism(s) involved in agonist-induced Cai2+ signaling is a potentially important molecular target for controlling angiogenesis and tumor growth. Several studies have shown that blood vessels in tumors differ from normal vessels in their morphology, blood flow and permeability. We had previously reported a key role for arachidonic acid (AA)-mediated Ca2+ entry in the initial stages of tumor angiogenesis in vitro. In this study we assessed the mechanism involved in AA-induced EC migration. We report that TRPV4, an AA-activated channel, is differentially expressed in EC derived from human breast carcinomas (BTEC) as compared with ‘normal’ EC (HMVEC). BTEC display a significant increase in TRPV4 expression, which was correlated with greater Ca2+ entry, induced by AA or 4αPDD (a selective TRPV4 agonist) in the tumor-derived ECs. Wound-healing assays revealed a key role of TRPV4 in regulating cell migration of BTEC but not HMVEC. Knockdown of TRPV4 expression completely abolished AA-induced BTEC migration, suggesting that TRPV4 mediates the pro-angiogenic effects promoted by AA. Furthermore, pre-incubation of BTEC with AA induced actin remodeling and a subsequent increase in the surface expression of TRPV4. This was consistent with the increased plasma membrane localization of TRPV4 and higher AA-stimulated Ca2+ entry in the migrating cells. Together, the data presented herein demonstrate that: (1) TRPV4 is differentially expressed in tumor-derived versus ‘normal’ EC; (2) TRPV4 has a critical role in the migration of tumor-derived but not ‘normal’ EC migration; and (3) AA induces actin remodeling in BTEC, resulting in a corresponding increase of TRPV4 expression in the plasma membrane. We suggest that the latter is critical for migration of EC and thus in promoting angiogenesis and tumor growth.

show abstract

Multiple Roles of Protein Kinase A in Arachidonic Acid–Mediated Ca2+ Entry and Tumor-Derived Human Endothelial Cell Migration

Cited by 36 publications

References 41 publications

Store-Operated Ca2+ Entry Is Remodelled and Controls In Vitro Angiogenesis in Endothelial Progenitor Cells Isolated from Tumoral Patients

Store-Operated Ca2+ Entry Is Remodelled and Controls In Vitro Angiogenesis in Endothelial Progenitor Cells Isolated from Tumoral Patients

TRPM8 inhibits endothelial cell migration via a non-channel function by trapping the small GTPase Rap1

TRPV4 mediates tumor-derived endothelial cell migration via arachidonic acid-activated actin remodeling

Contact Info

Product

Resources

About