title: High-resolution analysis of 16q22.1 in breast carcinoma using DNA amplifiable probes (multiplex amplifiable probe hybridization technique) and immunohistochemistry.
abstract: Loss of the chromosomal material at 16q22.1 is one of the most frequent genetic aberrations found in both lobular and low-grade nonlobular invasive carcinoma of the breast, indicating the presence of a tumour suppressor gene (TSG) at this region in these tumours. However, the TSG (s) at the 16q22.1 in the more frequent nonlobular carcinomas is still unknown. Multiplex Amplifiable Probe Hybridisation (MAPH) is a simple, accurate and a high-resolution technique that provides an alternative approach to DNA copy-number measurement. The aim of our study was to examine the most likely candidate genes at 16q22.1 using MAPH assay combined with protein expression analysis by immunohistochemistry. We identified deletion at 16q22.1 that involves some or all of these genes. We also noticed that the smallest region of deletion at 16q22.1 could be delineated to a 3 Mb region centromeric to the P-cadherin gene. Apart from the correlation between E-cadherin protein expression and its gene copy number, no correlation was detected between the expression of E2F-4, CTCF, TRF2 or P-cadherin with their gene's copy number. In the malignant tissues, no significant loss or decrease of protein expression of any gene other than E-cadherin was seen in association with any specific tumour type. No expression of VE-cadherin or Ksp-cadherin was detected in the normal and/or malignant tissues of the breast in these cases. However, there was a correlation between increased nuclear expression of E2F-4 and tumours with higher histological grade (p = 0.04) and positive lymph node disease (p = 0.02), suggesting that it may have an oncogenic rather than a tumour suppressor role. The malignant breast tissues also showed abnormal cytoplasmic cellular localisation of CTCF, compared to its expression in the normal parenchymal cells. In conclusion, we have demonstrated that MAPH is a potential technique for assessment of genomic imbalances in malignant tissues. Although our results support E-cadherin as the TSG in invasive lobular carcinoma, they argue against the candidacy of E2F-4, CTCF, TRF2, P-cadherin, Ksp-cadherin and VE-cadherin as TSGs in breast cancer.
authors.affiliation: The Breast Unit, Department of Histopathology, Nottingham City Hospital, University of Nottingham, United Kingdom.
authors.forename: Emad A
authors.initials: EA
authors.lastname: Rakha
authors.forename: John A L
authors.initials: JA
authors.lastname: Armour
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Pinder
authors.forename: Claire E
authors.initials: CE
authors.lastname: Paish
authors.forename: Ian O
authors.initials: IO
authors.lastname: Ellis
mesh_terms: D015703:Antigens, CD; D001943:Breast Neoplasms / Q000235:genetics* / Q000378:metabolism; D000076246:CCCTC-Binding Factor; D015820:Cadherins / Q000378:metabolism; D002277:Carcinoma / Q000235:genetics*; D002869:Chromosome Aberrations; D002885:Chromosomes, Human, Pair 16; D004247:DNA / Q000235:genetics*; D017931:DNA Primers / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000378:metabolism; D050704:E2F4 Transcription Factor; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction; D012097:Repressor Proteins / Q000378:metabolism; D035341:Telomeric Repeat Binding Protein 2 / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D000099152:Cadherin 5
chemical_list: D015703:Antigens, CD; D000076246:CCCTC-Binding Factor; D015820:Cadherins; D004247:DNA; D017931:DNA Primers; D004268:DNA-Binding Proteins; D050704:E2F4 Transcription Factor; D012097:Repressor Proteins; D035341:Telomeric Repeat Binding Protein 2; D014157:Transcription Factors; D000099152:Cadherin 5; C114483:CDH16 protein, human; C000619697:CTCF protein, human