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Isolation of a Novel Gene, TSP50, by a Hypomethylated DNA Fragment in Human Breast Cancer¹

Molecular Oncology, Hematology/Oncology Medicine [L. Y., J. S., V. V., H-p. X.], Department of Surgery [D. D. R.], Department of Laboratories [J. B.], and Division of Gynecologic Oncology [J. L., D. G.], North Shore-Long Island Jewish Health System, New York University School of Medicine, Manhasset, New York 11030

A novel gene, testes-specific protease 50 (TSP50), was isolated from a human testes cDNA library by using a genomic DNA probe, BR50. BR50 was isolated by a modified representational difference analysis (RDA) technique due to its hypomethylated feature in a breast cancer biopsy. This altered DNA methylation status was also detected by BR50 in other breast and some ovarian cancer tissues. The TSP50 gene product is a homologue to several human proteases, which indicates that it may encode a protease-like protein. Northern analysis of 16 different types of normal human tissues suggests that TSP50 was highly and specifically expressed in human testes, which indicates that it might possess a unique biological function(s) in that organ. Methylation status analysis in normal human testes and other tissues showed a correlation between DNA methylation and gene expression. Most importantly, reverse transcription-PCR analysis of 18 paired breast cancer tissues found that in 28% of the cancer samples, the TSP50 gene was differentially expressed. The possibility that TSP50 may be an oncogene is presently under investigation.

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