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

• Published in: Urolithiasis Vol. 47; no. 3; pp. 225 - 234
• Main Authors: Wang, Zhu, Zhang, Jian-wen, Zhang, Ying, Zhang, Sheng-ping, Hu, Qi-yi, Liang, Hui
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.06.2019; Springer Nature B.V
• Subjects: Cell growth; Cytotoxicity; Disease; Genomes; Kidney stones; Kidneys; Kinases; Medical Biochemistry; Medicine; Medicine & Public Health; Nephrology; Original Paper; Signal transduction; Urine; Urology; LncRNA; RNA seq; Calcium oxalate monohydrate; Kidney stone
• ISSN: 2194-7228; 2194-7236
• DOI: 10.1007/s00240-018-1065-7

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.