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Putrescine-dependent Invasive Capacity of Rat Ascites Hepatoma Cells¹

Departments of Biochemistry [Y. A., J-i. K., H. I.] and Pedodontics [F. K., M. N.], School of Dentistry, University of Tokushima, 3 Kuramoto-cho, Tokushima 770, and Department of Tumor Biochemistry, Center for Adult Diseases, Osaka, 1 Nakamichi, Higashinari-ku, Osaka 537 [K. S., H. A.], Japan

The effects of inhibitors of polyamine synthesis on the invasive capacity of rat ascites hepatoma (LC-AH) cells were examined by in vitro assay of penetration of the LC-AH cells through a monolayer of calf pulmonary arterial endothelial (CPAE) cells. Pretreatment of LC-AH cells with -difluoromethylornithine (DFMO), an inhibitor of ornithine decarboxylase, before seeding them onto a CPAE cell monolayer and culturing them for 24 h in the absence of DFMO decreased the number of penetrating tumor cells time and dose dependently (about 35% of the maximal inhibition) without affecting their viability or proliferative activity. DFMO treatment caused a marked decrease in the intracellular level of putrescine but not of spermidine or spermine. The DFMO-induced decreases in invasive capacity and putrescine level were almost completely reversed by the addition of putrescine to the medium during pretreatment with DFMO or invasion assay but were not affected by exogenous spermidine or spermine. No change in the invasive capacity was observed when the CPAE cells were treated with DFMO and the LC-AH cells with methylglyoxal-bis(guanylhydrazone), an inhibitor of S-adenosylmethionine decarboxylase, which depressed the spermidine and spermine levels but increased the putrescine level in the LC-AH cells.

These results suggest that intracellular putrescine modulates the in vitro invasive capacity of LC-AH cells.

¹ Supported in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan and by a Grant-in-Aid from the Ministry of Health and Welfare, Japan for a Comprehensive 10-Year Strategy for Cancer Control.

² To whom requests for reprints should be addressed.

Received 4/20/92. Accepted 7/21/92.

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