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Analysis of PRL, PRL-R, TGFβ1, and TGFβ-RII Gene Expression in Normal and Neoplastic Breast Tissues After Laser Capture Microdissection

1999; Lippincott Williams & Wilkins; Volume: 7; Issue: 3 Linguagem: Inglês

10.1097/00129039-199909000-00004

1541-2016

Sara Kuecker, Long Jin, Elzbieta Kulig, Ghislaine L. Oudraogo, Patrick C. Roche, Ricardo V. Lloyd,

Bone Metabolism and Diseases

Laser capture microdissection (LCM) is a new technique that provides rapid and reliable procurement of specific cells for molecular and other types of analyses. We used LCM to analyze normal and neoplastic breast tissues by reverse transcriptase polymerase chain reaction (RT-PCR) for expression of transforming growth factor beta 1 (TGFβ1), TGFβ receptor II (TGFβ-RII), prolactin (PRL), and PRL receptor (PRL-R) to evaluate this approach and to examine the autocrine/paracrine effects of these proteins and their receptors on mammary cell growth. RT-PCR analyses showed expression of TGFβ and TGFβ-RII mRNAs by most normal and neoplastic breast tissues. PRL and PRL-R mRNAs were detected in more cases of breast carcinomas than in normal breast tissues. The connective tissue stroma in the normal breast also expressed PRL-R as well as TGFβ1 and TGFβ-RII. Immunohistochemical staining of frozen sections of breast carcinoma for PRL-R followed by LCM (immuno-LCM) showed that the heterogeneity of PRL-R protein expression in carcinoma cells was also present at the mRNA level. These results show that LCM is a rapid and reproducible method for the molecular analysis of gene expression of TGFβ and PRL and their receptors in breast tissues. The immuno-LCM technique provides another level of specificity in performing molecular analysis of immunophenotypically characterized cells. The finding of PRL-R mRNA expression by the connective tissues in normal breasts suggests a paracrine regulatory role of PRL produced by the mammary epithelial cells on the adjacent stromal cells.