Department of Urology, People's Hospital of Longhua, 38 Jianshe RD, Longhua, Shenzhen, 518109, Guangdong, People's Republic of China.
Urolithiasis. 2019 Jun;47(3):225-234. doi: 10.1007/s00240-018-1065-7. Epub 2018 Jun 15.
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.
为了研究草酸钙一水合物附着的 HK-2 细胞中 lncRNA 和 mRNA 的表达谱，并探讨关键 lncRNA 表达水平与肾损伤之间的关联。将 HK-2 细胞用草酸钙晶体悬浮液处理。通过 MTT 测定法确定草酸钙晶体对 HK-2 细胞生长的影响。提取总 RNA，并通过高通量转录组测序平台 HiSeq 2500 分析 lncRNA 和 mRNA 的表达谱。通过实时 PCR 验证鉴定的 lncRNAs 和 mRNAs 的谱，并通过 Gene Ontology 数据库和 KEGG 信号通路分析分析其潜在功能。草酸钙晶体在几分钟内附着在 HK-2 细胞表面，并表现出明显的细胞毒性。RNA 测序结果显示，用草酸钙晶体处理的 HK-2 细胞中有 25 个差异表达的 lncRNA，其中 9 个上调，16 个下调。实时 PCR 验证结果具有统计学意义（P<0.05）。草酸钙晶体对人肾小管上皮细胞中的 lncRNA 和 mRNA 表达有显著影响，可能在肾结石介导的肾损伤中起关键作用。
