KCa3.1 (IK) modulates pancreatic cancer cell migration, invasion and proliferation: anomalous effects on TRAM-34
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In the recent decades, ion channels became the focus of cancer biologists, as many channels are overexpressed in tumour tissue and functionally they are linked to abnormal cell behaviour with processes including apoptosis, chemo- and radioresistance, proliferation and migration. KCa3.1 is a Ca2+-activated K+ channel that plays a central role in tumour progression in many cancer types. Therefore, the aim of the present study was to investigate KCa3.1 expression in pancreatic cancer cells and assess possible implications to disease progression. Using qPCR technique, we found abundant expression of KCa3.1 in pancreatic cancer cell lines. Patch clamp measurements on MiaPaCa-2 cells revealed a Ca2+-activated K+ current that matched biophysical characteristics as described for KCa3.1. Moreover, the current was sensitive to the commonly used channel modulators TRAM-34, clotrimazole and DC-EBIO, and it was abolished following transient gene knockdown of KCa3.1. We utilized both pharmacology and RNAi to assess a possible role of the channel in tumour cell behaviour. We found that the channel supported MiaPaCa-2 cell proliferation. Using RNAi protocols, we also identified KCa3.1 as important entity in cell invasion. However, TRAM-34 had unexpected stimulatory effects on cell migration and invasion estimated in various assays. Moreover, TRAM-34 increased intracellular Ca2+. In conclusion, we found prominent functional expression of KCa3.1 in pancreatic cancer cells. We provide evidence that the channel has a key role in cell proliferation and for the first time identify KCa3.1 as important entity in PDAC cell migration. We further reveal anomalous effects of TRAM-34.
KeywordsKCa3.1 KCNN4 TRAM-34 Clotrimazole DC-EBIO Pancreatic ductal adenocarcinoma Migration
This work was supported by the Marie Curie Initial Training Network IonTraC (Grant Agreement No. 289648), by The Danish Council for Independent Research/Natural Sciences (DFF - 4002-00162) and the German Cancer Aid (Project No. 110261). We are grateful to Ph.D. student Marco Tozzi for helping out with Boyden chamber assays and to E.H. Larsen and E.K. Hoffmann for fruitful discussions. Technical assistance of Pernille Roshof and Sabine Mally is greatly appreciated.
Compliance with ethical standards
Conflicts of interest
None of the authors have any conflict of interests.
All experiments were carried out in compliance to the current laws of Denmark and the UK.
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