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Growth-inhibitory Effects of Epidermal Growth Factor and Overexpression of Its Receptors on Human Squamous Cell Carcinomas in Culture¹

Department of Cancer Cell Research, Institute of Medical Science, University of Tokyo, Shirokanedai, Minato-ku, Tokyo 108 [N. K., K. C., T. K.], and Department of Oral Surgery, Tokyo Medical and Dental University, Faculty of Dentistry, Yushima, Bunkyo-ku, Tokyo 110 [K. R., M. H., S. E.], Japan

The role of epidermal growth factor (EGF) and its receptors in human cancers was studied using 24 human cell cultures including 15 of squamous cell carcinoma (SCC) of the skin, oral cavity, and esophagus. EGF was found to inhibit the growth and colony formation of all the SCC cells at doses that are mitogenic in many other cells, including epidermal keratinocytes and dermal fibroblasts. This inhibitory effect of EGF on SCCs was specific, because EGF did not inhibit and in some cases slightly stimulated the growth of other tumor cells, such as adenocarcinomas of the stomach, cervix, and breast and sarcomas.

The amounts of EGF receptors on these SCC cells were measured by immunoprecipitation of labeled proteins with anti-EGF receptor polyclonal antibody and binding assay of membrane preparations using ¹²⁵I-EGF. Of 13 SCC cell cultures tested, all except 3 of esophageal SCC showed higher levels of EGF receptor than normal epidermal keratinocytes, which contain 1.5 x 10⁵ binding sites/cell. In general, SCCs of the skin and oral cavity had large amounts of EGF receptor on the order of 10⁶/cell, whereas the receptor of esophageal SCCs was on the order of 10⁵/cell. Some SCC cells had about twice as many EGF receptors as A431 cells. The values for the equilibrium dissociation constant (K_d) of these cells were on the order of nM. The sensitivity to the inhibitory effect of EGF correlated well with the elevated level of EGF receptors in 12 SCC cell lines, and higher significance was obtained when data on esophageal SCCs were excluded. The present observations suggest that EGF and EGF receptors play a role in the development of SCCs.

¹ Supported in part by a grant for Cancer Research from the Ministry of Education, Science and Culture of Japan.

² To whom requests for reprints should be addressed.

Received 9/17/85. Revised 1/ 2/86. Accepted 1/ 3/86.

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