A Small Proline-rich Protein Regulated by Vitamin A in Tracheal Epithelial Cells Is Induced in Lung Tumors

Abstract

In cell-free translations of RNA from primary cultures of pig trachea surface epithelial cells, we observed that a 20 kD proline-rich protein (sPRP) is induced during culturing (Biochem. Biophys. Res. Commun. 1990; 172:1304-1309). Subsequently, a cDNA encoding sPRP has been cloned from pig tracheal cell mRNA and sequenced. This cDNA shows a high similarity to cDNAs cloned from monkey tracheal cells cultured in vitamin A-free medium and from UV-irradiated human epidermal keratinocytes. Amino acid sequences from these cDNAs are exceptionally rich in proline, glutamine, cysteine, and lysine but contain no aromatic amino acids. Two repeats of 12 amino acids on the N-terminus are followed by multiple 8 amino acid repeats. When compared with monkey trachea and human keratinocyte cDNAs, the sPRP cDNA from pig trachea has an additional 24 bp nucleotide repeat. Antiserum raised to a synthetic peptide (23 amino acids) on the C-terminus of sPRP (C23-antiserum) reacted with the 20 kD sPRP in immunoprecipitations from cell-free translations. On Northern blot analysis, sPRP cDNA hybridized to RNAs of similar sizes in tracheal cells from cat, rabbit, and lamb. sPRP was not detected in tracheal cells that were cultured with 10(-9) M arotinoid. Since sPRP is considered a putative squamous cell differentiation marker, experiments using lung tumors were performed. sPRP mRNA levels were dramatically increased in squamous lung tumors that were induced by injecting hamsters with 4-(methylnitrosamino)- 1-(3-pyridyl)-1-butanone, a tobacco-specific nitrosamine. In situ hybridization with tissue sections prepared from these lung tumors revealed that cells around the keratin pearls contained high levels of sPRP mRNA.