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This study is aimed at exploring the mechanism by which the −254C>G single nucleotide polymorphism (SNP) on the transient receptor potential cation channel 6 ( TRPC6 ) gene promoter could increase its activation in steroid-resistant nephrotic syndrome children of China. Plasmids containing the  TRPC6  promoter region (with the −254C or G allele) were constructed and then transfected into human embryonic kidney (HEK) 293T cells and human podocytes. Luciferase assays were used to test the promoter activity in both cell lines with or without tumor necrosis factor- α  (TNF- α ) treatment, and chromatin immunoprecipitation-polymerase chain reaction (ChIP-PCR) analysis was used to verify the transcription factor that could bind to this mutant sequence. Luciferase results indicate that the activity of the mutant promoter was greater than that of the normal promoter of the  TRPC6  gene in both cell lines. We further predicted and verified that this variation was mediated by the nuclear factor kappa B (NF- κ B) subunit RELA, and TNF- α  significantly enhanced the transcription activity of  TRPC6  with the −254G allele. In conclusion, the −254C>G SNP is a gain-of-function variation of the  TRPC6  gene, and it is also an early and effective factor for predicting steroid-resistant nephrotic syndrome (SRNS) in Chinese children.

international journal of genomics

−254C&gt;G SNP in the<i>TRPC6</i>Gene Promoter Influences Its Expression via Interaction with the NF-<i>κ</i>B Subunit RELA in Steroid-Resistant Nephrotic Syndrome Children

−254C>G SNP in theTRPC6Gene Promoter Influences Its Expression via Interaction with the NF-κB Subunit RELA in Steroid-Resistant Nephrotic Syndrome Children