RNA-sequencing analysis of HepG2 cells treated with atorvastatin

• Published in: PloS one 2014-08, Vol.9 (8), p.e105836
• Main Authors: Stormo, Camilla, Kringen, Marianne K, Lyle, Robert, Olstad, Ole Kristoffer, Sachse, Daniel, Berg, Jens P, Piehler, Armin P
• Format: Article
• Language: English
• Subjects: Acids; Alternative splicing; Anticholesteremic Agents - pharmacology; Atorvastatin; Biochemistry; Bioinformatics; Biology and Life Sciences; Biosynthesis; Cholesterol; Clinical medicine; Exons; Experiments; Gene expression; Gene Expression - drug effects; Gene sequencing; Genes; Genomes; Genomics; Hep G2 Cells; Hepatocytes - drug effects; Hepatocytes - metabolism; Heptanoic Acids - pharmacology; Homeostasis; Hospitals; Humans; Kinases; Low density lipoprotein; Medicine; Medicine and Health Sciences; MyD88 protein; Pyrroles - pharmacology; Ribonucleic acid; RNA; RNA Splicing; Sequence Analysis, RNA; Statins; Sterols; Transcription; Transcriptome
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0105836

The cholesterol-lowering drug atorvastatin is among the most prescribed drug in the world. Alternative splicing in a number of genes has been reported to be associated with variable statin response. RNA-seq has proven to be a powerful technique for genome-wide splice variant analysis. In the present study, we sought to investigate atorvastatin responsive splice variants in HepG2 cells using RNA-seq analysis to identify novel candidate genes implicated in cholesterol homeostasis and in the statin response. HepG2 cells were treated with 10 µM atorvastatin for 24 hours. RNA-seq and exon array analyses were performed. The validation of selected genes was performed using Taqman gene expression assays. RNA-seq analysis identified 121 genes and 98 specific splice variants, of which four were minor splice variants to be differentially expressed, 11 were genes with potential changes in their splicing patterns (SYCP3, ZNF195, ZNF674, MYD88, WHSC1, KIF16B, ZNF92, AGER, FCHO1, SLC6A12 and AKAP9), and one was a gene (RAP1GAP) with differential promoter usage. The IL21R transcript was detected to be differentially expressed via RNA-seq and RT-qPCR, but not in the exon array. In conclusion, several novel candidate genes that are affected by atorvastatin treatment were identified in this study. Further studies are needed to determine the biological significance of the atorvastatin responsive splice variants that have been uniquely identified using RNA-seq.