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Expression of the Second Calcitonin/Calcitonin Gene-Related Peptide Gene in Ewing Sarcoma Cell Lines^*

Institute of Molecular Biology, State University of Utrecht Utrecht, The Netherlands
Laboratory for Physiological Chemistry, State University of Utrecht Utrecht, The Netherlands
The Department of Internal Medicine of the University Hospital, State University of Utrecht Utrecht, The Netherlands
International Agency for Research on Cancer Lyon, France
The Research Laboratory for Calcium Metabolism, Departments of Orthopedic Surgery and Medicine, University of Zurich Zurich, Switzerland

Address all correspondence and requests for reprints to: Dr. J. W. M. Höppener, Institute of Molecular Biology, University of Utrecht, Transitorium III, Padualaan 8, Utrecht 3584 CH, The Netherlands.

The existence of two closely related calcitonin (CT)/CT gene-related peptide (CGRP) genes has been recognized in rat and man. In man, expression of the CALC-I gene produces CT and/or CGRP-I mRNA, whereas CGRP-II mRNA is transcribed from the CALC-II gene. Until recently, expression of the CALC-II gene had been detected only in a metastasis of medullary thyroid carcinoma. In this study, expression of the CALC-II gene was demonstrated by Northern blot hybridization analysis in four of six cell lines established from different Ewing sarcomas, a malignant neoplasm of bone. Expression of the CALC-I gene was not detected in any of the six cell lines. A presumed large mol wt immunoreactive precursor of CGRP-II and small amounts of mature CGRP-II, but no CT, were found in medium from IARC/EW 1 cells. Nucleotide sequence analysis of cloned cDNA from this cell line confirmed the production of CGRP-II mRNA. CALC-II gene expression in Ewing sarcoma might be useful for studies concerning regulation of gene expression in the CALC gene family and possibly for tumor classification.

^* This work was supported by the Swiss National Science Foundation (Grant 3.957-0.84), the Kanton of Zurich, and Ciba-Geigy, Ltd. (Basel, Switzerland).

Received July 29, 1986.

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