Analyses of long non-coding RNA and mRNA profiling using RNA sequencing in calcium oxalate monohydrate-stimulated renal tubular epithelial cells

Abstract

To study the expression profiles of lncRNA and mRNA in the calcium oxalate monohydrate-attached HK-2 cells, and investigate the association between critical lncRNA expression level and renal injury. The HK-2 cells were treated with crystal suspension of calcium oxalate. The effects of calcium oxalate crystals on the growth of HK-2 cells were determined by MTT assay. Total RNA was extracted and the lncRNA and mRNA expression profiles were analyzed by high-throughput transcriptase sequencing platform HiSeq 2500. The profile of identified lncRNAs and mRNAs were verified by real-time PCR and their potential function was analyzed by Gene Ontology database and KEGG signal pathway analysis. Calcium oxalate crystals adhered to the surface of HK-2 cells in few minutes and showed obvious cytotoxicity. RNA seq results showed that there were 25 differentially expressed lncRNAs in HK-2 cells treated with calcium oxalate crystals, of which 9 were up-regulated and 16 were down-regulated. The difference was verified by real-time PCR which showed statistically significant (P < 0.05). Calcium oxalate crystals have a significant effect on lncRNA and mRNA expression in human renal epithelial cells, which may play critical roles in kidney stone-mediated renal injury.

Keywords: Calcium oxalate monohydrate; Kidney stone; LncRNA; RNA seq.