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Thy-1 as a Negative Growth Regulator in ras-transformed Mouse Fibroblasts¹

Aging Process Research Laboratory (Frontier Research Program) [Y. S.], Laboratory of Molecular Oncology [Y. I.], and Laboratory of Molecular Regulation of Aging (Frontier Research Program) [H. N.], Tsukuba Life Science Center, The Institute of Physical and Chemical Research (RIKEN), Koyadai, Tsukuba, Ibaraki 305, Japan

To identify molecules on the cell surface involved in negative growth regulation, we assumed that their amounts would be reduced after malignant transformation. We analyzed several proteins by fluorescence-activated cell sorter in mouse NIH 3T3 and its transformed cell lines. Surprisingly, the amount of Thy-1, a cell surface glycoprotein anchored in the cell membrane by a glycophosphatidyl inositol linkage, was significantly decreased in the transformed NIH 3T3 lines, especially in ras-transformed NIH 3T3 lines. The malignant properties of clones of NIH 3T3 transformed by Kirsten murine sarcoma virus have a good correlation not only with the high amount of RAS proteins but also inversely with the amount of Thy-1. NIH 3T3 subpopulations lacking Thy-1 exhibit more susceptibility to the induction of colony-forming ability in soft agar by Kirsten murine sarcoma virus than the Thy-1-positive populations. Finally the transfection of Thy-1 complementary DNA to the ras-transformed NIH 3T3 significantly inhibits the colony formation in soft agar as well as the tumor formation in nude mice. Our results suggest that Thy-1 has negative effects on the anchorage-independent growth of ras-transformed NIH 3T3 cells.

¹ This work was partly supported by grants from the International Frontier Research Program and the Science and Technology Agency, Japan.

² To whom requests for reprints should be addressed.

Received 1/29/90. Accepted 9/25/90.

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