Figure 5. Polymorphisms at 8q24.21 influence expression of MYC and PVT1.

(a) HIF stabilization leads to allelic imbalance in PVT1 expression. Twelve primary renal tubular cell cultures (PTC) were identified that are heterozygous for SNP rs11604 that resides in the coding region of PVT1. Four individuals had an AC/AC genotype and eight individuals had an AC/CG genotype at rs35252396. For rs11604, the ratio of allele-specific signals was measured by qPCR (FAM/VIC) from input genomic DNA and cDNA derived from control (untreated) or DMOG-treated cells. No significant allelic imbalance at rs11604 was detected between the two groups (AC/AC or AC/CG at rs35252396) in input genomic DNA or cDNA from untreated cells (Supplementary Fig. 14). We then calculated the change of allelic ratios of cDNA from DMOG-treated cells compared with the respective control untreated cDNA. We detected a significant greater change in allelic PVT1 expression induced by DMOG in cells from individuals with an AC/CG genotype at rs35252396. Values are mean±s.d. from four (AC/AC) or eight (AC/CG) individuals. qPCRs for DNA and cDNA were performed in triplicates per individual. *Student's t-test, P<0.05. (b) Genotype expression correlation for rs10111989 and MYC in the KIRC TCGA data. rs10111989 is in LD with rs35252396 (r²=0.33, D′=0.98) and allele C is associated with RCC development (odds ratio 1.16, P<0.05) in data from a meta-analysis of UK and NCI cohorts⁵⁴. *χ²-test; P<0.05 for higher expression in CC individuals compared with TT individuals. Whiskers extend to 1.5 times of the inter quartile range. (c) Schematic representation of the 8q24.21 RCC enhancer of MYC and PVT1 expression. In cells from renal tubular origin (non-transformed tubular cells or RCC cells), HIFs can bind to the enhancer and regulate MYC and PVT1 expression, but binding is dependent on the rs35252396 genotype that affects accessibility of the site.