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Assessment of TRPV4-mediated calcium influx induced by receptor activation in MDA-MB-468 breast cancer cells

Citation

Armitage, K and Azimi, I and McIntyre, P and Bunnett, N and Roberts-Thomson, SJ and Monteith, GR, Assessment of TRPV4-mediated calcium influx induced by receptor activation in MDA-MB-468 breast cancer cells, ASCEPT-MPGPCR Joint Scientific Meeting 2016: Therapeutic targeting: right place, right time, right effect, 27-30 November 2016, Melbourne, Australia, pp. 26. (2016) [Conference Extract]


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Abstract

Introduction: The calcium permeable ion channel TRPV4 is involved in breast cancer cell invasion, migration and metastasis and appears to be associated with poorer patient outcomes. TRPV4 transactivation by G-protein coupled receptors and receptor tyrosine kinases, have been identified as important mechanisms of regulation in other tissues and pathologies. Characterisation of these transactivation pathways present an opportunity to better understand the nature of TRPV4 signalling in breast cancer cells.

Aims: To assess the sensitivity of Ca2+ influx elucidated by ATP, trypsin and epidermal growth factor (EGF) to the pharmacological TRPV4 inhibitor HC067047 and the Gαq inhibitor UBO-QIC in MDA-MB-468 breast cancer cells.

Methods: MDA-MB-468 cells were plated at 6 x 103 cells per well into 384 well black plates. For the measurement of free cytosolic Ca2+, cells were loaded with 2 μM PBX no-wash calcium indicator dye for 60 min at 37C, and were subsequently treated with TRPV4 and/or Gαq inhibitors for 15 min at room temperature. Intracellular Ca2+ levels associated with ATP, trypsin and epidermal growth factor (EGF) were then assessed using a Fluorescence Imaging Plate Reader (FLIPR).

Results: Ca2+ influx induced by the purinergic receptor activator ATP was unaffected by TRPV4 pharmacological inhibition. However, TRPV4 inhibition attenuated Ca2+ influx associated with trypsin-mediated activation of protease activated receptor 2 (PAR2) and EGF. Calcium influx induced by trypsin was UBO-QIC sensitive; in the presence of the Gαq inhibitor, TRPV4 inhibition had no effect on Ca2+ influx.

Discussion: These experiments suggest that PAR2 activation promotes Ca2+ influx in MDA-MB-468 breast cancer cells through a Gαq-dependent mechanism. Further investigation using siRNA silencing of TRPV4 and further assessment of the mechanistic components responsible for TRPV4 activation in breast cancer cells is now required.

Item Details

Item Type:Conference Extract
Keywords:TRPV4, calcium, breast cancer
Research Division:Biomedical and Clinical Sciences
Research Group:Pharmacology and pharmaceutical sciences
Research Field:Basic pharmacology
Objective Division:Expanding Knowledge
Objective Group:Expanding knowledge
Objective Field:Expanding knowledge in the health sciences
UTAS Author:Azimi, I (Dr Iman Azimi)
ID Code:151228
Year Published:2016
Deposited By:Pharmacy
Deposited On:2022-07-25
Last Modified:2022-07-28
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