title: DNA methylation profiling of phyllodes and fibroadenoma tumours of the breast.
abstract: Phyllodes tumours and cellular fibroadenomas are both fibroepithelial tumours of the breast. Phyllodes tumours, unlike fibroadenomas, have the ability to recur and metastasise. Although these lesions can be distinguished by their stromal cellularity, mitotic index, presence or absence of stromal overgrowth and cellular atypia, there is overlap and not infrequently a definitive diagnosis cannot be made, particularly on biopsy. We sought to evaluate whether DNA promoter methylation profiling using selected genes known to be methylated in cancer would allow us to learn more about the biology of these tumours, and whether it could identify methylation markers that could differentiate phyllodes tumours from fibroadenomas and/or distinguish phyllodes tumours of different grades. Methylation-sensitive high resolution melting (MS-HRM) was used to screen promoter DNA methylation changes in 86 phyllodes tumours (15 benign, 28 borderline, 43 malignant) and 26 fibroadenomas. A panel of 11 genes (RASSF1A, TWIST1, APC, WIF1, MGMT, MAL, RARβ, CDKN2A, CDH1, TP73 and MLH1) was tested. Methylation status was correlated with histology and with clinicopathological parameters. Five of the gene promoters showed some methylation in a proportion of phyllodes tumours; RASSF1A, 45.3%; TWIST1, 10.7%; APC, 4.1%; WIF1, 2.9% and MGMT, 1.3%. Only two genes showed any methylation in fibroadenomas usually at background levels; RASSF1A, 53.8% and MGMT, 8.3%. No CDKN2A methylation was observed in either tumour type, contrary to previous reports. Overall, the methylation patterns differed little from that which might be seen in normal cells. However, significant levels of methylation of RASSF1A (24.4%) and TWIST1 (7.1%) was observed in some phyllodes tumours. Elevated RASSF1A and/or TWIST1 methylation was significantly associated with phyllodes tumours compared with fibroadenomas (P = 0.02), TWIST1 methylation correlated with increasing malignancy in phyllodes tumours (P < 0.001). In conclusion, assessment of methylation of RASSF1A and TWIST1 may aid in the diagnosis of phyllodes tumours. The absence of frequent methylation in fibroadenomas supports a non-neoplastic origin.
authors.affiliation: Molecular Pathology Research and Development Laboratory, Department of Pathology, Peter MacCallum Cancer Centre, Melbourne, VIC, 3002, Australia.
authors.forename: Katie T
authors.initials: KT
authors.lastname: Huang
authors.forename: Alexander
authors.initials: A
authors.lastname: Dobrovic
authors.forename: Max
authors.initials: M
authors.lastname: Yan
authors.forename: Rooshdiya Z
authors.initials: RZ
authors.lastname: Karim
authors.forename: C Soon
authors.initials: CS
authors.lastname: Lee
authors.forename: Sunil R
authors.initials: SR
authors.lastname: Lakhani
authors.forename: Stephen B
authors.initials: SB
authors.lastname: Fox
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D001315:Australia; D014408:Biomarkers, Tumor / Q000235:genetics*; D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology; D016009:Chi-Square Distribution; D019175:DNA Methylation; D005260:Female; D018226:Fibroadenoma / Q000235:genetics* / Q000473:pathology; D020869:Gene Expression Profiling / Q000379:methods; D005819:Genetic Markers; D006801:Humans; D016015:Logistic Models; D008875:Middle Aged; D009687:Nuclear Proteins / Q000235:genetics; D003557:Phyllodes Tumor / Q000235:genetics* / Q000473:pathology; D011401:Promoter Regions, Genetic; D018570:Risk Assessment; D012307:Risk Factors; D025521:Tumor Suppressor Proteins / Q000235:genetics; D051879:Twist-Related Protein 1 / Q000235:genetics; D055815:Young Adult
chemical_list: D014408:Biomarkers, Tumor; D005819:Genetic Markers; D009687:Nuclear Proteins; C411762:RASSF1 protein, human; C103173:TWIST1 protein, human; D025521:Tumor Suppressor Proteins; D051879:Twist-Related Protein 1