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  • Title: Laser microdissection allows detection of abnormal gene expression in cystic adenomatoid malformation of the lung.
    Author: Jancelewicz T, Nobuhara K, Hawgood S.
    Journal: J Pediatr Surg; 2008 Jun; 43(6):1044-51. PubMed ID: 18558180.
    Abstract:
    BACKGROUND/PURPOSE: Congenital cystic adenomatoid malformation (CCAM) of the lung may result from a localized aberrant epithelial-mesenchymal interaction during lung development. We used laser microdissection (LMD) to isolate the epithelium and mesenchyme of CCAM, and studied candidate gene expression in these pure cell populations. METHODS: Congenital cystic adenomatoid malformation tissue was obtained from fetal (n = 5) and postnatal (n = 5) surgical specimens. Normal fetal lung (n = 10) was obtained from abortus material, and normal postnatal lung (n = 5) was identified from surgical specimens. Whole tissue was analyzed using immunohistochemistry and reverse transcriptase polymerase chain reaction (RT-PCR). Using LMD, columnar bronchiolar type epithelium and underlying mesenchyme were isolated. Multiplex nested RT-PCR was then used to detect message levels of candidate genes. RESULTS: Reverse transcriptase polymerase chain reaction performed on LMD-isolated tissue, but not whole tissue homogenate, revealed differences between CCAM and normal lung. In this report, we focus on the fibroblast growth factor (FGF) family. By RT-PCR, there was 4-fold more epithelial expression of FGF9 in fetal CCAM vs normal fetal lung (P < .07). This was qualitatively confirmed by immunohistochemistry. We also detected decreased FGF7 expression in CCAM mesenchyme (P < .05) but no significant differences in FGF10 or FGFR2. CONCLUSIONS: LMD may be used to overcome the limitations of tissue heterogeneity in the study of CCAM. Abnormal growth factor expression may play a role in the etiology of this lesion.
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