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Journal of Histochemistry and Cytochemistry, Vol. 47, 863-870, July 1999, Copyright © 1999, The Histochemical Society, Inc.

ARTICLE

Expression of REG Protein During Cell Growth and Differentiation of Two Human Colon Carcinoma Cell Lines

Françoise R. Bernard–Perronea, Wanda P. Renaudb, Odette M. Guy–Crotteb, Patrice Bernarda, Catherine G. Figarellab, Hiroshi Okamotoc, Daniel C. Balasa, and Françoise O. Senegas–Balasa
a Laboratoire d'Histologie, Faculté de Médecine, Nice, France
b Groupe de Recherche sur les Glandes Exocrines, Faculté de Médecine, Marseille, France
c Department of Biochemistry, Tohoku University School of Medecine, Miyagi, Japan

Correspondence to: Françoise O. Senegas–Balas, Laboratoire d’Histologie, Faculté de Médecine, av. de Valombrose, 06107 Nice Cedex 2, France.

We localized REG protein in Paneth cells and nonmature columnar cells of the human small intestinal crypts and speculated that this protein was associated with growth and/or differentiation. The aim of this study was to determine whether REG protein is present in two human colon cancer cell lines that exhibit enterocytic differentiation after confluence and to investigate changes in the level of its expression during growth and differentiation. Results were compared to those obtained on cells that remain undifferentiated. Western immunoblotting and immunofluorescence demonstrated the presence of REG protein in the three cell lines. With the antisera against human REG protein, the staining was diffusely spread throughout the cytoplasm at Day 2, and after Days 3–4 it appeared to have migrated to cell boundaries. After confluence, we observed only a punctate staining array along cell boundaries, which disappeared at Day 15. REG mRNA expression was demonstrated by RTPCR and REG mRNA hybridization until Day 13, but not after, in the three cell types. REG protein may be involved in cellular junctions. Its presence appears to be associated with the cell growth period and the protein must be downregulated when growth is achieved and differentiation is induced. (J Histochem Cytochem 47:863–870, 1999)

Key Words: REG protein, Caco-2, HT-29, growth, differentiation, human


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