title: [Expression and promoter methylation of SIX2 gene in peripheral blood of pediatric patients with nephroblastoma].
abstract: OBJECTIVE
  To explore the transcriptional expression and promoter methylation status of SIX2 gene in peripheral blood of pediatric children with nephroblastoma and discuss their clinicopathological correlations.
  
  
  METHODS
  Approved by the hospital ethics committee, peripheral blood samples were collected from 45 children with Wilms' tumor(case group) at the Department of Pediatric Surgery, First Affiliated Hospital, Zhengzhou University from October 2008 to January 2012. And another 15 pediatric cases gender-and-age matched, were selected as the control group (excluding cancer and other malignant diseases). The real-time quantitative (qRT)-PCR and methylation-specific PCR (MSP) were used to detect the mRNA expression level and methylation status of SIX2 gene.t or χ(2) test were used. Then analyzed their clinicopathological correlations in the case group and how SIX2 gene methylation affected its transcription.
  
  
  RESULTS
  Relative quantity(RQ) of SIX2 mRNA in the case group was higher than that of the control group (1.93 ± 1.10 vs 0.57 ± 0.39, t = 5.354, P = 0.000). There were 8 SIX2 gene methylation-positive cases in the case group versus 12 cases in the control group. And the methylation positive ratio was extremely lower in the case group (χ(2) = 19.600, P = 0.000). RQ values in the case group was associated with tumor size, clinical stage, pathological type, lymph node metastasis, treatment and outcome (all P < 0.05). RQ values in the methylated group was lower than that of the unmethylated group both in case and control group (1.35 ± 0.44 vs 1.95 ± 1.15, 0.43 ± 0.29 vs 1.13 ± 0.20, t = 2.459 and 3.896, P = 0.020 and 0.002) . RQ values of case group was higher than that of the control group in methylated group (t = 5.624, P = 0.000) . No statistical significance existed in RQ values between the case and control groups of unmethylated group (t = 1.222, P = 0.229) .
  
  
  CONCLUSIONS
  A close correlation between SIX2 low methylation and high mRNA expression in blood suggests that aberrant promoter methylation is possibly one of gene expression regulations, and may be correlated with the occurrence and development of Wilms' tumor. And SIX2 gene in methylated Wilms' tumor may play the role of oncogenes. A negative correlation exists between the overexpression in transcriptional level and its methylation status.
authors.affiliation: Department of Pediatric Surgery, First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China.
authors.forename: Dong-jian
authors.initials: DJ
authors.lastname: Song
authors.forename: Li-fang
authors.initials: LF
authors.lastname: Yue
authors.forename: He-ying
authors.initials: HY
authors.lastname: Yang
authors.forename: Xin-ya
authors.initials: XY
authors.lastname: Jia
authors.forename: Jun-jie
authors.initials: JJ
authors.lastname: Zhang
authors.forename: Yan-lei
authors.initials: YL
authors.lastname: Xu
authors.forename: Fei
authors.initials: F
authors.lastname: Guo
authors.forename: Jia-xiang
authors.initials: JX
authors.lastname: Wang
mesh_terms: D016022:Case-Control Studies; D002675:Child, Preschool; D019175:DNA Methylation; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D018398:Homeodomain Proteins / Q000097:blood* / Q000235:genetics; D006801:Humans; D007223:Infant; D007680:Kidney Neoplasms / Q000097:blood* / Q000235:genetics; D008297:Male; D009419:Nerve Tissue Proteins / Q000097:blood* / Q000235:genetics; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D009396:Wilms Tumor / Q000097:blood* / Q000235:genetics
chemical_list: D018398:Homeodomain Proteins; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; C484762:SIX2 protein, human