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High Invasiveness Associated with Augmentation of Motility in a fos-transferred Highly Metastatic Rat 3Y1 Cell Line¹

Department of Experimental Cell Research, Medical Institute of Bioregulation [S. T., K. N., M. I., H. S., T. B.], and Division of Orthopedic Surgery, Faculty of Medicine [H. I.], Kyushu University, Maidashi 3-1-1, Higashi-ku, Fukuoka 812; Department of Oncogene Research, Research Institute for Microbial Diseases, Osaka University, Suita 565 [M. T.]; and New Hoechst Building, No. 10-16, 8-chome, Akasaka, Minami-ku, Tokyo [H. O.], Japan

We previously reported that v-fos transfer to a src-transformed rat 3Y1 cell line enhanced lung metastasis. To clarify the mechanism of this enhancement, we compared various biological factors related to metastatic potential between a fos-transferred highly metastatic cell line (fos-SR-3Y1-202) and the control cell line transferred with genetic marker (pSV2-neo) plus pBR322, neo-SR-3Y1-200. Lung arrest, the effect of lung extract on cell growth, or sensitivity to natural killer cells could not explain the higher metastasis of fos-SR-3Y1-202, compared to findings with neo-SR-3Y1-200. The invasiveness, assessed by penetration through a Matrigel-coated filter was about 5 times higher in fos-SR-3Y1-202 than in neo-SR-3Y1-200; high invasiveness in vitro was also observed in a fos-transferred mixed-population cell line (fos-SR-3Y1-200) and fos-transferred highly metastatic clones. Histopathological evidence of an in vivo tumor also showed the high invasiveness of fos-SR-3Y1-202 cells. To elucidate the causes of the increased invasiveness of fos-SR-3Y1-202, attachment of the cells to Matrigel and its components, such as laminin and type IV collagen, type IV collagenase activity, and motility were examined. Attachment of the cells to the substrate coated on Petri dishes or the activity of type IV collagenase did not differ significantly. On the other hand, cell motility, determined by a new method to directly quantitate alteration of cell shape continuously, using video image analysis and computor techniques, was higher in fos-SR-3Y1-202 than in neo-SR-3Y1-200. Thus, the fos-transferred cell line, fos-SR-3Y1-202 has a high invasiveness, in association with augmentation of motility, hence the enhancement in metastatic potential.

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

² To whom requests for reprints should be addressed.

³ Present address: Department of Molecular Bioregulation, Institute of Molecular and Cellular Biology for Pharmaceutical Sciences, Kyoto Pharmaceutical University, Yamashina, Kyoto 607, Japan.

Received 3/20/89. Revised 8/28/89. Accepted 9/ 1/89.

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