Zhu Zewu, Huang Fang, Xia Weiping, Zeng Huimin, Gao Meng, Li Yongchao, Zeng Feng, He Cheng, Chen Jinbo, Chen Zhiyong, Li Yang, Cui Yu, Chen Hequn
Department of Urology, Xiangya Hospital, Central South University, Changsha, China.
Front Cell Dev Biol. 2021 Jan 11;8:596363. doi: 10.3389/fcell.2020.596363. eCollection 2020.
The current belief is that Randall's plaques (RP) constitute a nidus for the formation of idiopathic calcium oxalate stones, but the upstream events in RP formation remain unclear. The present study aimed to investigate whether RP formation shares similarities with biomineralization and to illustrate the potential role played by the lncRNA in osteogenic differentiation of human renal interstitial fibroblasts (hRIFs).
Biomineralization and expression were assessed in RP, and hRIFs were isolated and induced under osteogenic conditions for further experiments. The transcription initiation and termination sites in were identified by 5' and 3' RACE. RNA immunoprecipitation assays and luciferase assays were used to validate the interactions among , and miRNAs.
Upregulated expression of osteogenic markers and was observed in RP and hRIFs induced with osteogenic medium. Biomineralization in RP and calcium phosphate (CaP) deposits in induced hRIFs were further verified by electron microscopy. Furthermore, overexpression of promoted the osteogenic phenotype of hRIFs, while treatment with a miR-320a-5p mimic and knockdown of significantly suppressed the osteogenic phenotype. Further analysis showed that functioned as a competing endogenous RNA to sponge miR-320a-5p, leading to upregulation of Runx2 and thus promoting osteogenic differentiation of hRIFs.
Ectopic calcification and partially contributed to the formation of RP, in which might promote Runx2 expression to regulate osteogenic differentiation of hRIFs by sponging miRNA-320a-5p. The current study sheds new light on the lncRNA-directed mechanism of RP formation via a process driven by osteogenic-like cells.
目前的观点认为,兰德尔斑(RP)是特发性草酸钙结石形成的病灶,但RP形成的上游事件仍不清楚。本研究旨在探讨RP形成是否与生物矿化有相似之处,并阐明长链非编码RNA(lncRNA)在人肾间质成纤维细胞(hRIFs)成骨分化中所起的潜在作用。
评估RP中的生物矿化和lncRNA表达,并分离hRIFs并在成骨条件下诱导以进行进一步实验。通过5'和3'RACE确定lncRNA中的转录起始和终止位点。采用RNA免疫沉淀试验和荧光素酶试验验证lncRNA、微小RNA(miRNAs)之间的相互作用。
在RP和用成骨培养基诱导的hRIFs中观察到成骨标志物和lncRNA的表达上调。通过电子显微镜进一步证实了RP中的生物矿化以及诱导的hRIFs中的磷酸钙(CaP)沉积。此外,lncRNA的过表达促进了hRIFs的成骨表型,而用miR-320a-5p模拟物处理和lncRNA敲低则显著抑制了成骨表型。进一步分析表明,lncRNA作为竞争性内源RNA吸附miR-320a-5p,导致Runx2上调,从而促进hRIFs的成骨分化。
异位钙化和lncRNA部分促成了RP的形成,其中lncRNA可能通过吸附miRNA-320a-5p促进Runx2表达来调节hRIFs的成骨分化。本研究通过类成骨细胞驱动的过程,为RP形成的lncRNA导向机制提供了新的见解。