Down-Regulation of CD9 Expression during Prostate Carcinoma Progression Is Associated with CD9 mRNA Modifications

¹ Authors' Affiliations: ¹McMaster University Medical Centre, Hamilton, Ontario, Canada; ²Departments of Pathology and Surgery (Division of Urology), McGill University, Hôpital du Sacré-Coeur de Montréal; ³Division of Urology, Department of Surgery, McGill University; ⁴The Research Institute of the McGill University Health Centre, Montreal, Quebec, Canada; ⁵University of Western Ontario, London, Ontario, Canada; and ⁶Ecopia BioSciences, Inc. Ville Saint-Laurent, Quebec, Canada

^* To whom correspondence should be addressed. E-mail: mario.chevrette{at}mcgill.ca.

	Abstract

Purpose: Cluster-of-differentiation antigen 9 (CD9) protein, a member of the tetraspanin family, has been implicated in carcinogenesis of various human tumors. Although decreased expression of the CD82 tetraspanin protein, a close CD9 relative, is associated with prostate cancer progression, CD9 expression has not been analyzed in this malignancy.

Experimental Design: CD9 expression in human prostatic adenocarcinoma was analyzed by immunohistochemistry on 167 primary tumors and 88 lymph node or bone metastases. CD9 cDNA was sequenced from two human prostate cancer cell lines, prostatic adenocarcinoma, high-grade prostatic intraepithelial neoplasia (PIN), and normal prostatic tissues.

Results: Although CD9 was detected in the epithelium of normal prostatic tissues, reduced or loss of CD9 expression within neoplastic cells was observed in 24% of 107 clinically localized primary adenocarcinomas, 85% of 60 clinically advanced primary adenocarcinomas, 85% of 65 lymph node metastases, and 65% of 23 bone metastases. Difference in CD9 expression between clinically localized and advanced diseases was highly significant (P < 1 x 10^-7). Whereas there was no alteration of CD9 cDNA in normal tissues, all PC-3-derived cell lines, one PIN, and four prostatic adenocarcinomas harbored deletions in their CD9 cDNAs. Recurring CD9 point mutations were also found in PC-3M-LN4 cells, one PIN, and seven prostatic adenocarcinomas.

Conclusions: CD9 expression is significantly reduced and even lost during prostate cancer progression. Moreover, deletions and mutations of the CD9 mRNA may be associated with loss of protein expression observed in tumor cells. Our data suggest that CD9 inactivation may play an important role in prostate cancer progression.

Key Words: CD9, prostate carcinoma, prostatic adenocarcinoma, prostatic intraepithelial neoplasia (PIN), transmembrane 4 superfamily (TM4SF), mRNA deletions and mutations, RNA editing