title: Methylation of tumor suppressor gene CDH13 and SHP1 promoters and their epigenetic regulation by the UHRF1/PRMT5 complex in endometrial carcinoma.
abstract: OBJECTIVE
  Epigenetic changes in cancer and precancerous lesions could be utilized as biomarkers for cancer early detection. This study aims to investigate the novel biomarkers in endometrial carcinoma, and explore their epigenetic regulation.
  
  
  METHODS
  Methylation of six tumor suppressor genes (CDH13, SHP1, HIN1, DKK3, CTNNA1 and PCDH8) was evaluated in 155 endometrium samples. Changes of methylation and mRNA expression after treatment with 5-Aza-2'-deoxycytidine (5-Aza-CdR) or/and trichostatin A (TSA) were investigated by MSP and qRT-PCR respectively. Co-immunoprecipitation was used to detect the interactions between UHRF1 and PRMT5 proteins.
  
  
  RESULTS
  CDH13 and SHP1 promoters were highly methylated (81.36% and 86.44%, respectively) in endometrial carcinoma, while CDH13 promoter methylation was also present in complex hyperplasia and atypical hyperplasia (51.72% and 50.00%, respectively). Methylation of CDH13 and SHP1 promoters was associated with age and tumor differentiation or muscular infiltration depth. CDH13 and SHP1 promoters were completely methylated in endometrial carcinoma cell lines and were partially reversed by 5-Aza-CdR or TSA to induce mRNA levels (P<0.01). After combined treatment with these two agents, methylation of CDH13 and SHP1 promoters was completely reversed and expression of their mRNA was significantly increased (P<0.01). Moreover, PRMT5 could bind to UHRF1 and down-regulated by 5-Aza-CdR and/or TSA treatment (P<0.05).
  
  
  CONCLUSIONS
  Our data demonstrate for the first time that SHP1 methylation has high specificity for diagnosis of endometrial carcinoma, while CDH13 promoter methylation plays a role in the earlier stage. Furthermore, UHRF1 could form a complex with PRMT5 to contribute to the endometrial carcinogenesis.
authors.affiliation: Institute of Pathology and Pathophysiology, Shandong University School of Medicine, Jinan 250012, China; Department of Gynecology and Obstetrics, Tianjin Medical University General Hospital, Tianjin, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Sheng
authors.affiliation: Tianjin Medical University Cancer Institute and Hospital, National Clinical Research for Cancer, The Key Laboratory of Cancer Prevention and Therapy, Tianjin 300060, China.
authors.forename: Hongtao
authors.initials: H
authors.lastname: Wang
authors.affiliation: Institute of Pathology and Pathophysiology, Shandong University School of Medicine, Jinan 250012, China.
authors.forename: Dongchen
authors.initials: D
authors.lastname: Liu
authors.affiliation: Institute of Pathology and Pathophysiology, Shandong University School of Medicine, Jinan 250012, China.
authors.forename: Cheng
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Institute of Pathology and Pathophysiology, Shandong University School of Medicine, Jinan 250012, China.
authors.forename: Yupeng
authors.initials: Y
authors.lastname: Deng
authors.affiliation: Institute of Pathology and Pathophysiology, Shandong University School of Medicine, Jinan 250012, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Yang
authors.affiliation: Institute of Pathology and Pathophysiology, Shandong University School of Medicine, Jinan 250012, China. Electronic address: ztguo@sdu.edu.cn.
authors.forename: Tingguo
authors.initials: T
authors.lastname: Zhang
authors.affiliation: Institute of Pathology and Pathophysiology, Shandong University School of Medicine, Jinan 250012, China. Electronic address: cuijuanzhang@sdu.edu.cn.
authors.forename: Cuijuan
authors.initials: C
authors.lastname: Zhang
mesh_terms: D001374:Azacitidine / Q000031:analogs & derivatives / Q000494:pharmacology; D022762:CCAAT-Enhancer-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D015820:Cadherins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D019175:DNA Methylation; D000077209:Decitabine; D016889:Endometrial Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D005260:Female; D016147:Genes, Tumor Suppressor; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D008875:Middle Aged; D011401:Promoter Regions, Genetic; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6 / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D011484:Protein-Arginine N-Methyltransferases / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D044767:Ubiquitin-Protein Ligases; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D022762:CCAAT-Enhancer-Binding Proteins; D015820:Cadherins; C099972:H-cadherin; D006877:Hydroxamic Acids; D012333:RNA, Messenger; C012589:trichostatin A; D000077209:Decitabine; C400529:PRMT5 protein, human; D011484:Protein-Arginine N-Methyltransferases; C406364:UHRF1 protein, human; D044767:Ubiquitin-Protein Ligases; C509249:PTPN6 protein, human; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6; D001374:Azacitidine
keywords: DNA methylation; Endometrial carcinoma; Epigenetics; Tumor suppressor genes