Suppression of Tumorigenicity by the Wild-Type Tuberous Sclerosis 2 (Tsc2) Gene and its C-Terminal Region

• Published in: Proceedings of the National Academy of Sciences - PNAS 1996-08, Vol.93 (17), p.9154-9159
• Main Authors: Jin, Fang, Wienecke, Ralf, Xiao, Guang-Hui, Maize, John C., DeClue, Jeffrey E., Yeung, Raymond S.
• Format: Article
• Language: English
• Subjects: Animals; Carcinoma - etiology; Carcinoma - genetics; Carcinoma - pathology; Cell growth; Cell Line; Cell lines; Disease Models, Animal; Fibroblasts; Genes, Tumor Suppressor; Kidney cells; Kidney Neoplasms - etiology; Kidney Neoplasms - genetics; Kidney Neoplasms - pathology; Kidneys; Mice; Mice, SCID; Mutation; Peptide Fragments - biosynthesis; Peptide Fragments - genetics; Plasmids; Rats; Repressor Proteins - biosynthesis; Repressor Proteins - genetics; Transfection; Tuberous sclerosis; Tuberous Sclerosis - etiology; Tuberous Sclerosis - genetics; Tuberous Sclerosis - pathology; Tuberous Sclerosis Complex 2 Protein; Tumor cell line; Tumor Suppressor Proteins; Tumors
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.93.17.9154

The Tsc2 gene, which is mutationally inactivated in the germ line of some families with tuberous sclerosis, encodes a large, membrane-associated GTPase activating protein (GAP) designated tuberin. Studies of the the Eker rat model of hereditary cancer strongly support the role of Tsc2 as a tumor suppressor gene. In this study, the biological activity of tuberin was assessed by expressing the wild-type Tsc2 gene in tumor cell lines lacking functional tuberin and also in rat fibroblasts with normal levels of endogenous tuberin. The colony forming efficiency of Eker rat-derived renal carcinoma cells was significantly reduced following reintroduction of wild-type Tsc2. Tumor cells expressing the transfected Tsc2 gene became more anchorage-dependent and lost their ability to form tumors in severe combined immunodeficient mice. At the cellular level, restoration of tuberin expression caused morphological changes characterized by enlargement of the cells and increased contact inhibition. As with the full-length Tsc2 gene, a clone encoding only the C terminus of tuberin (amino acids 1049-1809, including the GAP domain) was capable of reducing both colony formation and in vivo tumorigenicity when transfected into the Eker rat tumor cells. In normal Rat1 fibroblasts, conditional overexpression of tuberin also suppressed colony formation and cell growth in vitro. These results provide direct experimental evidence for the tumor suppressor function of Tsc2 and suggest that the tuberin C terminus plays an important role in this activity.