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mRNA expression profiling of phyllodes tumours of the breast: identification of genes important in the development of borderline and malignant phyllodes tumours
Author(s) -
Jones AM,
Mitter R,
Poulsom R,
Gillett C,
Hanby AM,
Consortium Phyllodes Tumour,
Tomlinson IPM,
Sawyer EJ
Publication year - 2008
Publication title -
the journal of pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.964
H-Index - 184
eISSN - 1096-9896
pISSN - 0022-3417
DOI - 10.1002/path.2439
Subject(s) - immunohistochemistry , phyllodes tumor , gene expression profiling , cancer research , stromal cell , hmga2 , cell growth , gene , biology , gene expression , cell , pathology , medicine , genetics , microrna
The aim of this study was to identify genes involved in the development of borderline and malignant phyllodes tumours of the breast (PTs). Expression profiling of 23 PTs (12 benign, 11 borderline/malignant) was performed using Affymetrix U133A GeneChips. mRNA expression in the borderline/malignant PTs was compared to the benign PTs. A group of 162 genes was over‐expressed in the borderline/malignant group with a fold change > 2 and FDR < 0.1. Four of these genes were chosen for further investigation: PAX3, SIX1, TGFB2 and HMGA2 . Over‐expression was validated in a separate set of formalin‐fixed, paraffin‐embedded (FFPE) tumours, using either in situ hybridization or immunohistochemistry. This confirmed that expression of PAX3, SIX1, TGFB2 and HMGA2 in the stromal component of PTs was associated with the borderline/malignant phenotypes ( p = 8.7 × 10 −5 , p = 0.05, p = 0.009, p = 0.003, respectively; Fisher's exact test). The functional consequences of down‐regulating these genes were studied using siRNA in short‐term cultures and cell lines established from PTs. mRNA ‘knock‐down’ of PAX3 resulted in significantly decreased cell proliferation in both a malignant and a borderline PT cell culture. mRNA ‘knock‐down’ of SIX1 and HMGA2 resulted in decreased cell proliferation only in the malignant PT cell line, and ‘knock‐down’ of TGFB2 resulted in decreased cell proliferation only in the borderline PT cell culture. This study shows that these four genes are involved in the development of borderline/malignant PTs. SIX1 over‐expression was most marked in the highly malignant PTs, with particularly high expression in one case of metastatic PT. PAX3, TGFB2 and HMGA2 were expressed predominantly in borderline/malignant PTs, but showed some expression in benign tumours; they may be important in the transition from the benign to borderline/malignant phenotype. Copyright © 2008 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.