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MCF-10A Cells Infected with the int-2 Oncogene Induce Angiogenesis in the Chick Chorioallantoic Membrane and in the Rat Mesentery¹

Romano Danesi², Cristiana Agen, Antonello Di Paolo, Fulvio Basolo, Silvia Del Bianchi and Mario Del Tacca

Istituto di Farmacologia Medica, Università di Pisa, 56126 Pisa [M. C., C. A., M. D. T.]; Scuola Superiore di Studi Universitari e di Perfezionamento S. Anna, 56127 Pisa [R. D., A. D. P.]; Istituto di Anatomia e Istologia Patologica, Università di Pisa, 56126 Pisa [F. B.]; and Centro Ricerche Esperienze e Studi Applicazioni Militari, 56100 Pisa [S. D. B.], Italy

² To whom requests for reprints should be addressed, at Clinical Pharmacology Branch, Bldg. 10, Rm. 12C-103, National Cancer Institute, NIH, Bethesda, MD 20892.

A growing body of evidence demonstrates the relevant role of the int-2 (FGF-3) oncogene in human carcinomas. To investigate its angiogenic activity, the human epithelial mammary cell line MCF-10A was infected with a retroviral expression vector carrying the int-2 oncogene. Infected cells were entrapped in an alginate pellet and placed on the chorioallantoic membrane of chick embryos. After 7 days, a dense capillary network was found to grow toward the pellet, whereas parental cells did not show any angiogenic activity. Conditioned medium from int-2-infected cells was injected i.p. twice daily into rats over a period of 10 days. The mesentery of treated rats showed numerous small blood vessels originating from larger vascular arcades and growing through the stromal layer of the mesentery. In control experiments, neither medium for cell culture nor conditioned medium from parental cells was found to induce angiogenesis. In conclusion, the stimulation of blood vessel growth by int-2-infected cells suggests that the production of the int-2 protein is associated with the acquisition of the angiogenic phenotype.

¹ Supported in part by grants from the Italian Association for Cancer Research (Associazione Italiana Ricerca sul Cancro, Milano, Italy). Preliminary results were presented at the 84th Annual Meeting of the American Association for Cancer Research, Orlando, FL.

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Received 8/ 9/93. Accepted 11/12/93.

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