Objective
We have recently discovered that human prostate cancer cells possess anion membrane conductance that is sensitive to the changes in the cell volume. Considering potential modulation of volume-regulated anion channel (VRAC) by P-glycoprotein, that confers multi-drug resistance on human tumours, as well as its probable association with phospholemman-like protein that is highly expressed in tumours it is almost certain that this channel may be involved in prostate cancerogenesis. Therefore, it is very important to determine functional properties of VRAC in prostate cancer cells, to elucidate its molecular nature, to evaluate its role in cells growth and proliferation, and to compare membrane properties of normal and cancerous prostate epithelial cells. Even though we will use LNCaP (Lymph Node Carcinoma of the Prostate) cell line as a primary model for our study essential part of the experiments will be conducted on primary cells isolated from human tissue specimens obtained from prostate surgeries, which will provide clinical relevance to the proposed research.
The following specific aims will be pursued in this study:
To determine functional properties (voltage dependence, selectivity, pharmacology, single channel conductance, intracellular Ca2+-, ATP- and cAMP-sensitivity, role of intracellular ion strength) of volume-sensitive anion conductance in LNCP cells;
To determine if activation of this conductance can be affected by the state of cellular cytoskeleton and small G-proteins which are cytoskeletal organisers (e.g. RhoA);
To determine which known Cl- channel-related proteins - ClC-3, phospholemman, ClC-2, P-glycoprotein, pICln or any other underlie and/or modulate VRAC in prostate cancer cells with the placement of the main emphasis on the most probable candidates - ClC-3 and phospholemman;
To get insight into the signalling mechanisms involved in the gating of VRAC in prostate cancer cells e.g. to examine the role of PKA, PKC, tyrosine kinases and tyrosine phosphathases, PTK´s, PTP´s, and intracellular ionic strength in the VRAC modulation;
To examine how activation of VS anion conductance affects prostate cancer cells growth and proliferation.
Topic(s)
Call for proposal
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59655 Villeneuve d'Ascq Cedex
France