Allele-specific <em>CDH1</em> downregulation and hereditary diffuse gastric cancer

Hugo Pinheiro

Renata Bordeira-Carriço

Susana Seixas

Joana Carvalho

Jorge Rocha+3 authors

Supplementary Material

[Supplementary Data]

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^{Institute of Molecular Pathology and Immunology, University of Porto (IPATIMUP), Porto 4200-465, Portugal,}

^{Hereditary Cancer Program, British Columbia Cancer Agency, Vancouver, Canada, V5Z 4E6,}

^{Department of Zoology and Anthropology, Faculty of Sciences, University of Porto, Porto 4169-007, Portugal}

^{Faculty of Medicine, University of Porto, Porto 4200-319, Portugal}

^{To whom correspondence should be addressed at: IPATIMUP, Rua Roberto Frias s/n, 4200-465 Porto, Portugal. Tel: +351 225570700; Fax: +351 225570799; Email:}tp.pumitapi@loalrac

Received 2009 Aug 24; Revised 2009 Nov 10; Accepted 2009 Dec 1.

Abstract

Hereditary diffuse gastric cancer (HDGC) is an autosomal dominant cancer susceptibility syndrome characterized by early-onset diffuse gastric cancer (DGC) and lobular breast cancer. E-cadherin (CDH1) heterozygous germline mutations and deletions are found in 40% of families. Independent of CDH1 alterations, most HDGC tumours display mislocalized or absent E-cadherin immunoexpression, therefore undetected defects at the CDH1 locus may still be involved. We aimed at determining whether CDH1 mutation-negative probands display germline CDH1 allele-specific expression (ASE) imbalance, using a single-nucleotide primer extension-based procedure and tried to uncover the underlying molecular defect. CDH1 ASE analysis was performed using three intragenic SNPs in RNA extracted from the blood of 21 cancer-free individuals and 22 HDGC probands (5 CDH1 mutation carriers and 17 CDH1 negative). Germline promoter methylation, deletions and haplotype-related susceptibility at the CDH1 locus were analysed. Both CDH1 alleles from cancer-free individuals displayed equivalent expression levels, whereas monoallelic CDH1 expression or high allelic expression imbalance (AI) was present in 80% of CDH1 mutant and 70.6% (n = 12) of CDH1-negative HDGC probands. Germline deletions and promoter hypermethylation were found in 25% of probands displaying high CDH1 AI. No particular haplotype was found to be associated with CDH1 high AI. Germline CDH1 AI is highly frequent among CDH1 mutation-negative probands but was not seen in cancer-free individuals. This implicates the CDH1 locus in the majority of mutation-negative HDGC families.

Abstract

NA, not applicable; high A1 values and CDH1 alterations are highlighted in bold.

^{See Results section.}

^{Mutation negative and not tested for large deletions.}

^{Family 1.}

^{Family 4 in reference (}4).

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ACKNOWLEDGEMENTS

We gratefully acknowledge patients, families and their caregivers for their willing participation in this project and who provided consent regarding the use of the information obtained from the study. We thank Luis Maia, Marta Novais and Teresa Coelho for providing biological material from cancer-free individuals.

Conflict of Interest statement. None declared.

ACKNOWLEDGEMENTS

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