Application of differential display, with in situ hybridization verification, to microscopic samples of breast cancer tissue

Ruey Ho Kao, Giulio Francia, Richard Poulsom, Andrew M. Hanby, Ian R. Hart

研究成果: 雜誌貢獻文章

4 引文 (Scopus)

摘要

The technique of differential display (DD) has been used widely to identify potentially interesting overexpressed or repressed genes in a variety of compared samples. When used in studying tissue samples, it inevitably confronts problems of limited amount of input material and cell-type heterogeneity. We report here the application of in situ hybridization as a method of confirmatory test for DD as well as definition of cell type expressing differential cDNA. This procedure employed material derived from a single case of human mammary, grade III, infiltrating ductal carcinoma, using free-hand microdissection, where we have compared gene expression profiles in invasive tumour with those in adjacent normal tissue. A total of 21 cDNAs were found to be differentially expressed between the two tissue types; 11 upregulated in the tumour sample and 10 upregulated in the normal sample. Six cDNAs were utilized as probes for in situ hybridization analysis of a further five cases of comparably staged breast cancer. One of these clones, 11AT1, which was found to be homologous to Hsc70, was shown to be overexpressed in tumour cells relative to adjacent normal stroma and to benign glandular epithelium in all five cases; an increase in expression was further confirmed at protein level by immunohistochemistry. The study demonstrated the applicability of in situ hybridization as a screening test in DD strategy for studying tissue material and a reasonable technique combination of identifying changes in gene expression associated with tumour development.
原文英語
頁(從 - 到)207-212
頁數6
期刊International Journal of Experimental Pathology
84
發行號5
DOIs
出版狀態已發佈 - 十月 1 2003
對外發佈Yes

指紋

In Situ Hybridization
Breast Neoplasms
Complementary DNA
Neoplasms
Microdissection
Ductal Carcinoma
Transcriptome
Breast
Epithelium
Hand
Clone Cells
Immunohistochemistry
Gene Expression
Genes
Proteins

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Molecular Biology
  • Cell Biology

引用此文

Application of differential display, with in situ hybridization verification, to microscopic samples of breast cancer tissue. / Kao, Ruey Ho; Francia, Giulio; Poulsom, Richard; Hanby, Andrew M.; Hart, Ian R.

於: International Journal of Experimental Pathology, 卷 84, 編號 5, 01.10.2003, p. 207-212.

研究成果: 雜誌貢獻文章

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