Macrodissection versus microdissection of rectal carcinoma: minor influence of stroma cells to tumor cell gene expression profiles

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• 作者：Elza C de Bruin (1)
  Simone van de Pas (1)
  Esther H Lips (2)
  Ronald van Eijk (2)
  Minke MC van der Zee (1)
  Marcel Lombaerts (2)
  Tom van Wezel (2)
  Corrie AM Marijnen (1)
  J Han JM van Krieken (3)
  Jan Paul Medema (1)
  Cornelis JH de van Velde (4)
  Paul HC Eilers (5)
  Lucy TC Peltenburg (1)
  
• 刊名：BMC Genomics
• 出版年：2005
• 出版时间：December 2005
• 年：2005
• 卷：6
• 期：1
• 全文大小：351KB
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• 作者单位：Elza C de Bruin (1)
  Simone van de Pas (1)
  Esther H Lips (2)
  Ronald van Eijk (2)
  Minke MC van der Zee (1)
  Marcel Lombaerts (2)
  Tom van Wezel (2)
  Corrie AM Marijnen (1)
  J Han JM van Krieken (3)
  Jan Paul Medema (1)
  Cornelis JH de van Velde (4)
  Paul HC Eilers (5)
  Lucy TC Peltenburg (1)
  
  1. Department of Clinical Oncology, Leiden University Medical Center, Albinusdreef 2, 2333, ZA,Leiden, The Netherlands
  2. Department of Pathology, Leiden University Medical Center, Albinusdreef 2, 2333, ZA Leiden, The Netherlands
  3. Department of Pathology, University Medical Center St. Radboud, Geert Grooteplein-Zuid 10, 6525, GA Nijmegen, The Netherlands
  4. Department of Surgery, Leiden University Medical Center, Albinusdreef 2, 2333, ZA Leiden, The Netherlands
  5. Department of Medical Statistics, Leiden University Medical Center, Wassenaarseweg 62, 2333, AL,Leiden, The Netherlands
  

文摘

Background The molecular determinants of carcinogenesis, tumor progression and patient prognosis can be deduced from simultaneous comparison of thousands of genes by microarray analysis. However, the presence of stroma cells in surgically excised carcinoma tissues might obscure the tumor cell-specific gene expression profiles of these samples. To circumvent this complication, laser microdissection can be performed to separate tumor epithelium from the surrounding stroma and healthy tissue. In this report, we compared RNAs isolated from macrodissected, of which only surrounding healthy tissue had been removed, and microdissected rectal carcinoma samples by microarray analysis in order to determine the most reliable approach to detect the expression of tumor cell-derived genes by microarray analysis. Results As microdissection yielded low tissue and RNA quantities, extra rounds of mRNA amplification were necessary to obtain sufficient RNA for microarray experiments. These second rounds of amplification influenced the gene expression profiles. Moreover, the presence of stroma cells in macrodissected samples had a minor contribution to the tumor cell gene expression profiles, which can be explained by the observation that more RNA is extracted from tumor epithelial cells than from stroma. Conclusion These data demonstrate that the more convenient procedure of macrodissection can be adequately used and yields reliable data regarding the identification of tumor cell-specific gene expression profiles.