title: TWIST1 and TWIST2 promoter methylation and protein expression in tumor stroma influence the epithelial-mesenchymal transition-like tumor budding phenotype in colorectal cancer.
abstract: Tumor budding in colorectal cancer is likened to an epithelial-mesenchymal transition (EMT) characterized predominantly by loss of E-cadherin and up-regulation of E-cadherin repressors like TWIST1 and TWIST2. Here we investigate a possible epigenetic link between TWIST proteins and the tumor budding phenotype. TWIST1 and TWIST2 promoter methylation and protein expression were investigated in six cell lines and further correlated with tumor budding in patient cohort 1 (n = 185). Patient cohort 2 (n = 112) was used to assess prognostic effects. Laser capture microdissection (LCM) of tumor epithelium and stroma from low- and high-grade budding cancers was performed. In colorectal cancers, TWIST1 and TWIST2 expression was essentially restricted to stromal cells. LCM results of a high-grade budding case show positive TWIST1 and TWIST2 stroma and no methylation, while the low-grade budding case was characterized by negative stroma and strong hypermethylation. TWIST1 stromal cell staining was associated with adverse features like more advanced pT (p = 0.0044), lymph node metastasis (p = 0.0301), lymphatic vessel invasion (p = 0.0373), perineural invasion (p = 0.0109) and worse overall survival time (p = 0.0226). Stromal cells may influence tumor budding in colorectal cancers through expression of TWIST1. Hypermethylation of the tumor stroma may represent an alternative mechanism for regulation of TWIST1.
authors.affiliation: Translational Research Unit (TRU), Institute of Pathology, University of Bern, Bern 3010, Switzerland.
authors.forename: José A
authors.initials: JA
authors.lastname: Galván
authors.affiliation: Translational Research Unit (TRU), Institute of Pathology, University of Bern, Bern 3010, Switzerland.
authors.forename: Melina
authors.initials: M
authors.lastname: Helbling
authors.affiliation: Translational Research Unit (TRU), Institute of Pathology, University of Bern, Bern 3010, Switzerland.|Clinical Pathology Division, Institute of Pathology, University of Bern, Switzerland.
authors.forename: Viktor H
authors.initials: VH
authors.lastname: Koelzer
authors.affiliation: Experimental Pathology Division, Institute of Pathology, University of Bern, Switzerland.
authors.forename: Mario P
authors.initials: MP
authors.lastname: Tschan
authors.affiliation: Department of Medical Oncology, Bern University Hospital, Bern, Switzerland.
authors.forename: Martin D
authors.initials: MD
authors.lastname: Berger
authors.affiliation: Departments of Visceral Surgery and Medicine, Bern University Hospital, Bern, Switzerland.
authors.forename: Marion
authors.initials: M
authors.lastname: Hädrich
authors.affiliation: Departments of Visceral Surgery and Medicine, Bern University Hospital, Bern, Switzerland.
authors.forename: Beat
authors.initials: B
authors.lastname: Schnüriger
authors.affiliation: Translational Research Unit (TRU), Institute of Pathology, University of Bern, Bern 3010, Switzerland.|Clinical Pathology Division, Institute of Pathology, University of Bern, Switzerland.
authors.forename: Eva
authors.initials: E
authors.lastname: Karamitopoulou
authors.affiliation: Translational Research Unit (TRU), Institute of Pathology, University of Bern, Bern 3010, Switzerland.|Clinical Pathology Division, Institute of Pathology, University of Bern, Switzerland.
authors.forename: Heather
authors.initials: H
authors.lastname: Dawson
authors.affiliation: Departments of Visceral Surgery and Medicine, Bern University Hospital, Bern, Switzerland.|Department of Surgery, Tiefenau Hospital, Bern, Switzerland.
authors.forename: Daniel
authors.initials: D
authors.lastname: Inderbitzin
authors.affiliation: Translational Research Unit (TRU), Institute of Pathology, University of Bern, Bern 3010, Switzerland.|Clinical Pathology Division, Institute of Pathology, University of Bern, Switzerland.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Lugli
authors.affiliation: Translational Research Unit (TRU), Institute of Pathology, University of Bern, Bern 3010, Switzerland.
authors.forename: Inti
authors.initials: I
authors.lastname: Zlobec
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015820:Cadherins / Q000378:metabolism; D015331:Cohort Studies; D015179:Colorectal Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D019175:DNA Methylation; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics*; D005260:Female; D045325:HCT116 Cells; D019073:HT29 Cells; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D053208:Kaplan-Meier Estimate; D008297:Male; D008875:Middle Aged; D015999:Multivariate Analysis; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D011379:Prognosis; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism; D017422:Sequence Analysis, DNA / Q000379:methods; D017154:Stromal Cells / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D051879:Twist-Related Protein 1 / Q000235:genetics* / Q000378:metabolism; D000071799:Zinc Finger E-box-Binding Homeobox 1; D051176:beta Catenin / Q000378:metabolism
chemical_list: D015820:Cadherins; D018398:Homeodomain Proteins; D009687:Nuclear Proteins; D012097:Repressor Proteins; C103173:TWIST1 protein, human; C486274:TWIST2 protein, human; D014157:Transcription Factors; D051879:Twist-Related Protein 1; C486307:ZEB1 protein, human; D000071799:Zinc Finger E-box-Binding Homeobox 1; D051176:beta Catenin