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Identification of Calcitonin and Calcitonin Gene-Related Peptide Messenger Ribonucleic Acid in Medullary Thyroid Carcinomas by Hybridization Histochemistry^*

Department of Medicine, University of Melbourne, Repatriation General Hospital Heidelberg 3081
Howard Florey Institute of Experimental Medicine and Physiology Parkville 3052 Australia

Address requests for reprints to: Dr. T. J. Martin, Department of Medicine, University of Melbourne, Repatriation General Hospital, Heidelbert 3081, Australia.

Synthesis and secretion of calcitonin and calcitonin gene-related peptide (CGRP) were studied in medullary thyroid carcinomas (MTC) by hybridization histochemistry on tissue sections and by Northern gel analyis of mRNA. Five patients with MTC and elevated serum levels of calcitonin and CGRP were studied. Surgically obtained tumor samples (four primary and three lymph node metastases) were extracted after freezing, and the RNA was fractionated on Northern gels. Hybridization was carried out with ³²P-labeled synthetic oligodeoxyribonucleotides coding specifically for calcitonin and CGRP. Calcitonin- and CGRP-specific mRNAs approximately 1000 nucleotides in length were demonstrated in all 7 tumor samples. However, neither calcitonin nor CGRP mRNA was detected in a pheochromocytoma from 1 of the patients who had multiple endocrine neoplasia type II. A series of unselected lung carcinomas yielded the same result. Hybridization histochemistry was carried out on sections from the same tumors using the same probes. The mRNAs for calcitonin and CGRP were located in all cells of neoplastic MTC appearance, with CGRP mRNA at significantly lower levels. This demonstrated that both calcitonin and CGRP mRNA were present within the same tumor cells. The lung tumors and pheochromocytoma were negative with both probes. Hybridization histochemistry is likely to be of use in diagnosis of medullary thyroid cancer and in studying the calcitonin-CGRP mRNA processing mechanism in whole cells.

^* This work was supported by grants from the National Health and Medical Research Council of Australia.

Received July 29, 1985.

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