Silencing of TRPC1 regulates store-operated calcium entry and proliferation in Huh7 hepatocellular carcinoma cells

WOS: 000354179500030PubMed ID: 25960236Purpose: Previously, we observed reciprocal changes in TRPC1 and TRPC6 expression levels in aging rat aorta and A7r5, rat embryonic vascular smooth muscle cells. Furthermore, downregulation of TRPC1 significantly elevated store-operated Ca2+ entry suggesting the regulatory role of TRPC1 in A7r5 cells. Since TRPC6 upregulation shown to be associated with cell proliferation, the purpose of our study was to investigate the functional consequences of TRPC1 ion channel downregulation by RNA interference in Huh7 human hepatocellular carcinoma cell line. Methods: Huh7 cells used in quantitative gene and protein expression as well as in functional analyses. To determine mRNA and protein levels, quantitative real-time RT-PCR and western blot analyses were performed, respectively. In functional analyses, real-time changes in proliferation, migration and intracellular Ca2+ levels were monitored. Results: In shTRPC1-transfected Huh7 cells, TRPC1 mRNA and protein levels significantly decreased whereas store-operated Ca2+ entry significantly elevated. TRPC1-silencing suppressed cell proliferation without affecting cell migration in real-time cellular analyses. Conclusion: These results suggest that TRPC1 may take part both in regulation of store-operated Ca2+ entry and proliferation of hepatocellular carcinoma cells. (C) 2015 Elsevier Masson SAS. All rights reserved.Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [108S072, BIDEB-2211]; Ege UniversityEge University [EBILTEM-09BIL003, BAP-06ECZ013]; NovartisNovartisThis work was supported by the Scientific and Technological Research Council of Turkey (TUBITAK) Research Project [108S072] to Tosun M.; and Graduate Scholarship [BIDEB-2211] to Selli C. A partial support was also provided by the Ege University [EBILTEM-09BIL003; BAP-06ECZ013] and; Novartis [Pharmacology Research Project-2008] to Tosun M. to conduct initial experiments. The authors would like to thank Professor M. Ozturk for providing Huh7 cells and plasmid vectors