Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
8538698 | Toxicology and Applied Pharmacology | 2018 | 40 Pages |
Abstract
HL1-hT1 cell line represents adult human liver stem cells (LSCs) immortalized with human telomerase reverse transcriptase. In this study, HL1-hT1 cells were found to express mesenchymal markers (vimentin, CD73, CD90/THY-1 and CD105) and an early hepatic endoderm marker FOXA2, while not expressing hepatic progenitor (HNF4A, LGR5, α-fetoprotein) or differentiated hepatocyte markers (albumin, transthyretin, connexin 32). In response to microcystin-LR (MC-LR), a time- and concentration-dependent formation of MC-positive protein bands in HL1-hT1 cells was observed. Cellular accumulation of MC-LR occurred most likely via mechanisms independent on organic anion transporting polypeptides (OATPs) or multidrug resistance (MDR) proteins, as indicated (a) by a gene expression analysis of 11 human OATP genes and 4 major MDR genes (MDR1/P-glycoprotein, MRP1, MRP2 and BCRP); (b) by non-significant effects of OATP or MDR1 inhibitors on MC-LR uptake. Accumulation of MC-positive protein bands in HL1-hT1 cells was associated neither with alterations of cell viability and growth, dysregulations of ERK1/2 and p38 kinases, reactive oxygen species formation, induction of double-stranded DNA breaks nor modulations of stress-inducible genes (ATF3, HSP5). It suggests that LSCs might have a selective, MDR1-independent, survival advantage and higher tolerance towards MC-induced cytotoxic, genotoxic or cancer-related events than differentiated adult hepatocytes, fetal hepatocyte or malignant liver cell lines. HL1-hT1 cells provide a valuable in vitro tool for studying effects of toxicants and pharmaceuticals on LSCs, whose important role in the development of chronic toxicities and liver diseases is being increasingly recognized.
Keywords
TTRRT-PCRMicrocystinsDAPIPAGERESIPSCsBSPreserpineVimentinBcrpDSBsMDR1OATPCx32OATPsVIMCFDA-AMLSCsDNA double-stranded breakstransthyretinICCMC-LRhESCsCX43SDSALBMCs4′,6-diamidino-2-phenylindoleHCChTERTMAPKsROSα-fetoproteinFOCAlamar BlueAlbuminpolyacrylamide gel electrophoresisImmunocytochemistrybromosulfophthaleinionizing radiationtaurocholateAFPhuman telomerase reverse transcriptaseNeutral redRifampicinsodium dodecyl sulfatehuman embryonic stem cellsInduced pluripotent stem cellsMenmenadionemicrocystin-LRreverse transcriptase polymerase chain reactionWestern blottingbreast cancer resistance proteinmultidrug resistance protein 1Multidrug resistance proteinsOrganic anion transporting polypeptidesHepatocellular carcinomaConnexin 32Rifconnexin 43mitogen-activated protein kinasesReactive oxygen species
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Authors
Jan Raska, Lucie Ctverackova, Aneta Dydowiczova, Iva Sovadinova, Ludek Blaha, Pavel Babica,