Abstract:
Objective To explore the molecular regulatory mechanism of long non-coding RNA H19 and aarF domain containing kinase 4 (ADCK4) gene and provide new insights into diagnosing and treating nephrotic syndrome in children.
Methods The blood samples from 30 children with nephrotic syndrome and 30 controls were collected from Wuxi Children's Hospital Affiliated to Jiangnan University. The expressions of H19 and ADCK4 were detected by real-time quantitative polymerase chain reaction (qRT-PCR). Also the expression of ADCK4 was detected by qRT-PCR or Western blot (WB) when H19 was over-expressed or knocked down in podocytes. Luciferase activity was quantified whether H19 could regulate the promoter activity of ADCK4. Furthermore, mass spectrometry was employed for identifying the transcription factor of binding with H19 and RNA immunoprecipitation assay (RIPA) for confirming the interaction between H19 and candidate transcription factor.
Results The expressions of H19 and ADCK4 became down-regulated in peripheral blood mononuclear cells (PBMCs). Overexpression of H19 promoted the expression of ADCK4 in podocytes while H19 knockdown suppressed it. Furthermore, our study demonstrated that H19 could regulate the promoter activity of ADCK4. Using RNA pull-down and mass spectrometry revealed that transcription factor THAP1 (THAP domain-containing protein 1) could bind with H19 and mutual interaction was further confirmed by RIPA.
Conclusions H19 provides new rationales for diagnosing and treating nephrotic syndrome in children.
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