Isolation, characterization, and expression of feline stromelysin-1 in naturally developing tumors in cats

Kara C. Sorensen Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois, Urbana, IL 61801.
Present address is Department of Natural Resources and Environmental Sciences, College of Agricultural, Consumer, and Environmental Sciences, University of Illinois, Urbana, IL 61802.

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Barbara E. Kitchell Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois, Urbana, IL 61801.
Present address is Department of Small Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824-1314.

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Arthur M. Siegel Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois, Urbana, IL 61801.

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Paul Mardis Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois, Urbana, IL 61801.

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Abstract

Objective—To detect, isolate, and characterize feline stromelysin-1 (ie, matrix metalloproteinase [MMP]-3) in naturally developing tumors in cats.

Sample Population—31 tissue samples obtained from primary tumors and 6 samples of normal tissues from cats.

Procedure—Biopsy specimens were obtained from primary tumors. Primers were designed on the basis of known sequences. The sequence of stromelysin- 1 was cloned and analyzed. An additional primer set was used as a screening tool. Samples were assayed in duplicate or triplicate, when possible. Data obtained were analyzed for differences in expression of stromelysin-1 with regard to overall survival among cats of various sex, age, and disease status.

Results—A 1,181-bp cDNA nucleotide sequence was amplified. The open reading frame encoded 393 amino acids. This amino acid sequence shared 70% to 85% sequence homology with sequences of other species. In addition, samples were screened for stromelysin-1. Of the 31 tumor samples tested, 16 (51.6%) had positive results for expression of stromelysin-1. Total RNA expression was detected in a diverse group of tumor types. Prognostic factors associated with a shorter duration of survival included evidence of metastasis and metastasis associated with expression of stromelysin-1.

Conclusions and Clinical Relevance—Feline stromelysin-1 contains all the conserved regions typically found in members of the MMP family. Activity of stromelysin-1 has been implicated in a wide number of physiologic and pathologic processes. Identification of this gene may lead to the development of useful reagents to assist with diagnosis and management of neoplastic diseases in cats. (Am J Vet Res 2004; 65:213–219)

Abstract

Objective—To detect, isolate, and characterize feline stromelysin-1 (ie, matrix metalloproteinase [MMP]-3) in naturally developing tumors in cats.

Sample Population—31 tissue samples obtained from primary tumors and 6 samples of normal tissues from cats.

Procedure—Biopsy specimens were obtained from primary tumors. Primers were designed on the basis of known sequences. The sequence of stromelysin- 1 was cloned and analyzed. An additional primer set was used as a screening tool. Samples were assayed in duplicate or triplicate, when possible. Data obtained were analyzed for differences in expression of stromelysin-1 with regard to overall survival among cats of various sex, age, and disease status.

Results—A 1,181-bp cDNA nucleotide sequence was amplified. The open reading frame encoded 393 amino acids. This amino acid sequence shared 70% to 85% sequence homology with sequences of other species. In addition, samples were screened for stromelysin-1. Of the 31 tumor samples tested, 16 (51.6%) had positive results for expression of stromelysin-1. Total RNA expression was detected in a diverse group of tumor types. Prognostic factors associated with a shorter duration of survival included evidence of metastasis and metastasis associated with expression of stromelysin-1.

Conclusions and Clinical Relevance—Feline stromelysin-1 contains all the conserved regions typically found in members of the MMP family. Activity of stromelysin-1 has been implicated in a wide number of physiologic and pathologic processes. Identification of this gene may lead to the development of useful reagents to assist with diagnosis and management of neoplastic diseases in cats. (Am J Vet Res 2004; 65:213–219)

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