Coordinate regulation by transcription factors and DNA methylation in the core promoter region of SIRT6 in bovine adipocytes.

Sirtuin6 (SIRT6) is an ADP-ribosyltransferase and NAD+-dependent deacylase of acetyl groups and long-chain fatty acyl groups, and has been shown as a regulator of insulin secretion, glucose metabolism, lipid metabolism, and cancer. In this study, we determined that the bovine SIRT6 showed higher levels of mRNA expression in the testis, longissimus thoracis, and subcutaneous fat tissue. To elucidate the molecular regulation mechanism of bovine SIRT6 expression, we obtained a 2-kb fragment containing the 5'-regulatory region, and the functional proximal minimal promoter of bovine SIRT6 was identified in the -472/-73 bp region. The CCAAT enhancer binding protein beta (CEBPß), paired box 6 (PAX6), Kruppel-like factor 2 (KLF2), myb proto-oncogene protein (CMYB), nuclear respiratory factor 1 (NRF1), and E2F transcription factor 1 (E2F1) binding sites, as transcriptional activators or repressors in the core promoter region of SIRT6, were determined by electrophoretic mobility shift assay (EMSA) experiments and luciferase reporter assays. In addition, the results from methylation assay and luciferase report assay showed that the bovine SIRT6 promoter activity was coordinately regulated by methylation and NRF1 or E2F1 during bovine adipocyte differentiation. Taken together, this study illuminated the underlying mechanism of methylation and transcription regulation of SIRT6 expression in bovine adipocytes.

[Distribution of WNT10A gene rs10177996 polymorphism between Han and Uygur populations in Xinjiang area].

PURPOSE: To explore the characteristics of distribution of WNT10A gene rs10177996 polymorphism between Han and Uygur populations in Xinjiang area. METHODS: A cross-sectional survey on 154 Han individuals in Urumqi area and 134 Uygur individuals in Kashgar area was performed. Buccal epithelial cells were harvested using Cotton swab scraping, and DNA was extracted by special kit. After screening, the corresponding SNP segments of qualified samples were propagated by PCR. Dideoxy-mediated chain termination method was used for gene sequencing, and then, genotyping was conducted with corresponding software. Statistical analysis of genetic data was performed by SPSS 23.0 software package. RESULTS: Among Uygur nationality in Kashgar area, the frequencies of CC, CT, TT genetypes in rs10177996 were 8.21%, 30.60% and 61.19%, respectively. The allele frequency of C was 23.51% and T was 76.49%. Among Han nationality in Urumqi area, the frequencies on CC, CT, TT genetypes of rs10177996 were 9.74%, 43.51% and 46.75%, respectively. The allele frequency of C was 31.49% and T was 68.51%. When compared with Han nationality, the frequency of TT was significantly higher in Uygur nationality(P=0.046). When compared with European, the frequency of TT was significantly lower in Uygur nationality (P=0.05). When compared with European, the frequency of TT was significantly lower in Han nationality(P＜0.01). Compared with European, the distribution on C allele frequency was significantly higher, the distribution on T allele frequency was significantly lower in Han nationality (P=0.033). However, there was no significant difference between Han nationality in Urumqi area and Uygur nationality in Kashgar area (P＞0.05), and, between Uygur nationality in Kashgar area and European (P＞0.05). Meanwhile, there was no significant difference in gender between Uygur nationality in Kashgar area and Han nationality in Urumqi area (P＞0.05). CONCLUSIONS: The distributions of WNT10A gene rs10177996 SNP among Han nationality in Urumqi area, Uygur nationality in Kashgar area and the reported European population are obviously different.