title: Silencing of TRPC1 regulates store-operated calcium entry and proliferation in Huh7 hepatocellular carcinoma cells.
abstract: OBJECTIVE
  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 Ca(2+) 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 Ca(2+) levels were monitored.
  
  
  RESULTS
  In shTRPC1-transfected Huh7 cells, TRPC1 mRNA and protein levels significantly decreased whereas store-operated Ca(2+) entry significantly elevated. TRPC1-silencing suppressed cell proliferation without affecting cell migration in real-time cellular analyses.
  
  
  CONCLUSIONS
  These results suggest that TRPC1 may take part both in regulation of store-operated Ca(2+) entry and proliferation of hepatocellular carcinoma cells.
authors.affiliation: Department of Pharmacology, Faculty of Pharmacy, Ege University, 35040 Izmir, Turkey. Electronic address: cigdem.selli@ege.edu.tr.
authors.forename: Cigdem
authors.initials: C
authors.lastname: Selli
authors.affiliation: Department of Pharmacology, Faculty of Pharmacy, Ege University, 35040 Izmir, Turkey. Electronic address: yasemin.erac@ege.edu.tr.
authors.forename: Yasemin
authors.initials: Y
authors.lastname: Erac
authors.affiliation: Department of Medical Biology, Faculty of Medicine, Ege University, 35040 Izmir, Turkey. Electronic address: buket.kosova@ege.edu.tr.
authors.forename: Buket
authors.initials: B
authors.lastname: Kosova
authors.affiliation: Department of Medical Biology and Genetics, Faculty of Medicine, Dokuz Eylul University, 35340 Izmir, Turkey. Electronic address: esra.erdal@deu.edu.tr.
authors.forename: Esra S
authors.initials: ES
authors.lastname: Erdal
authors.affiliation: Department of Pharmacology, Faculty of Pharmacy, Ege University, 35040 Izmir, Turkey. Electronic address: metiner.tosun@ege.edu.tr.
authors.forename: Metiner
authors.initials: M
authors.lastname: Tosun
mesh_terms: D002118:Calcium / Q000378:metabolism*; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000473:pathology*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D020868:Gene Silencing / Q000187:drug effects; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D042541:Intracellular Space / Q000378:metabolism; D008113:Liver Neoplasms / Q000235:genetics / Q000473:pathology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D050052:TRPC Cation Channels / Q000235:genetics / Q000378:metabolism*; D000074287:TRPC6 Cation Channel; D014162:Transfection
chemical_list: D007211:Indoles; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D050052:TRPC Cation Channels; D000074287:TRPC6 Cation Channel; C117032:TRPC6 protein, human; C096351:transient receptor potential cation channel, subfamily C, member 1; D002118:Calcium; C000543:cyclopiazonic acid
keywords: Calcium store depletion; Cell proliferation; Fura-2; TRP channels