Methylation in the promoter regions of WT1, NKX6-1 and DBC1 genes in cervical cancer tissues of Uygur women in Xinjiang

<div><p>Abstract This study aimed to explore: 1) DNA methylation in the promoter regions of Wilms tumor gene 1 (WT1), NK6 transcription factor related locus 1 gene (NKX6-1) and Deleted in bladder cancer 1 (DBC1) gene in cervical cancer tissues of Uygur women in Xinjiang, and 2) the correlation of gene methylation with the infection of HPV16/18 viruses. We detected HPV16/18 infection in 43 normal cervical tissues, 30 cervical intraepithelial neoplasia lesions (CIN) and 48 cervical cancer tissues with polymerase chain reaction (PCR) method. Methylation in the promoter regions of the WT1, NKX6-1 and DBC1 genes in the above-mentioned tissues was measured by methylation-specific PCR (MSP) and cloning sequencing. The expression level of these three genes was measured by real-time PCR (qPCR) in 10 methylation-positive cervical cancer tissues and 10 methylation-negative normal cervical tissues. We found that the infection of HPV16 in normal cervical tissues, CIN and cervical cancer tissues was 14.0, 36.7 and 66.7%, respectively. The infection of HPV18 was 0, 6.7 and 10.4%, respectively. The methylation rates of WT1, NKX6-1 and DBC1 genes were 7.0, 11.6 and 23.3% in normal cervical tissues, 36.7, 46.7 and 30.0% in CIN tissues, and 89.6, 77.1 and 85.4% in cervical cancer tissues. Furthermore, WT1, NKX6-1 and DBC1 genes were hypermethylated in the high-grade squamous intraepithelial lesion (CIN2, CIN3) and in the cervical cancer tissues with infection of HPV16/18 (both P< 0.05). The expression of WT1, NKX6-1 and DBC1 was significantly lower in the methylation-positive cervical cancer tissues than in methylation-negative normal cervical tissues. Our findings indicated that methylation in the promoter regions of WT1, NKX6-1 and DBC1 is correlated with cervical cancer tumorigenesis in Uygur women. The infection of HPV16/18 might be correlated with methylation in these genes. Gene inactivation caused by methylation might be related to the incidence and development of cervical cancer.</p></div

Additional file 1: of Variants of human papillomaviruses 16 (HPV16) in Uigur women in Xinjiang, China

Phylogenetic trees of HPV16 E6, E7 and L1 variants based on Neighbor Joining Molecular Phylogenetic analysis. Figure S1. Neighbor Joining Molecular Phylogenetic analysis using 43 nucleotide sequences of HPV16 E6 gene. Phylogenetic studies were performed on E6 nucleotide sequence alignment of 477 positions from each case, which was constructed by the neighbor joining method and the Kimura 2-Parameter model by MEGA 6 package. Bootstrap proportions were calculated with 1000 replicates. Study sequences are labeled in KT GenBank accession numbers, others are reference GenBank sequences. E, European variant; Ep, European prototype; As, Asia lineage; AA, Asian American lineage; Af, African lineage. Figure S2. Neighbor Joining Molecular Phylogenetic analysis using 43 nucleotide sequences of HPV16 E7 gene. Phylogenetic studies were performed on E7 nucleotide sequence alignment of 297 positions from each case, which was constructed by the neighbor joining method and the Kimura 2-Parameter model by MEGA 6 package. Bootstrap proportions were calculated with 1000 replicates. Study sequences are labeled in KT GenBank accession numbers, others are reference GenBank sequences. E, European variant; Ep, European prototype; As, Asia lineage; AA, Asian American lineage; Af, African lineage. Figure S3.  Neighbor Joining Molecular Phylogenetic analysis using 39 nucleotide sequences of HPV16 L1 gene. Phylogenetic studies were performed on partial L1 nucleotide sequence alignment of 369 positions from each case, which was constructed by the neighbor joining method and the Kimura 2-Parameter model by MEGA 6 package. Bootstrap proportions were calculated with 1000 replicates. Study sequences are labeled in KT GenBank accession numbers, others are reference GenBank sequences. E, European variant; Ep, European prototype; As, Asia lineage; AA, Asian American lineage; Af, African lineage. (PDF 136 kb

Supplementary figure 1 -Supplemental material for Nuclear factor kappa B regulated monocyte chemoattractant protein-1/chemokine CC motif receptor-2 expressing in spinal cord contributes to the maintenance of cancer-induced bone pain in rats

<p>Supplemental material, Supplementary figure 1 for Nuclear factor kappa B regulated monocyte chemoattractant protein-1/chemokine CC motif receptor-2 expressing in spinal cord contributes to the maintenance of cancer-induced bone pain in rats by Yungong Wang, Huadong Ni, Hongbo Li, Houshen Deng, Long S Xu, Shijie Xu, Ying Zhen, Hui Shen, Huan Pan and Ming Yao in Molecular Pain</p