Laser microdissection allows detection of abnormal gene expression in cystic adenomatoid malformation of the lung.

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.

• Authors: Tim Jancelewicz, Kerilyn Nobuhara, Sam Hawgood
• Journal: Journal of pediatric surgery (J Pediatr Surg) Vol. 43 Issue 6 Pg. 1044-51 (Jun 2008) ISSN: 1531-5037 [Electronic] United States
• PMID: 18558180 (Publication Type: Comparative Study, Journal Article)

Chemical References

• FGF10 protein, human
• Fibroblast Growth Factor 10
• Fibroblast Growth Factor 9
• Fibroblast Growth Factor 7
• Fibroblast Growth Factors

Topics

• Child
• Cohort Studies
• Cystic Adenomatoid Malformation of Lung, Congenital (genetics, pathology)
• Female
• Fetus (pathology)
• Fibroblast Growth Factor 10 (genetics)
• Fibroblast Growth Factor 7 (genetics)
• Fibroblast Growth Factor 9 (genetics)
• Fibroblast Growth Factors (genetics)
• Gene Expression Regulation, Developmental
• Genetic Predisposition to Disease
• Humans
• Immunohistochemistry
• In Situ Nick-End Labeling
• Infant
• Infant, Newborn
• Lasers
• Male
• Microdissection (instrumentation, methods)
• Reference Values
• Reverse Transcriptase Polymerase Chain Reaction
• Risk Assessment
• Sensitivity and Specificity
• Tissue Culture Techniques