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Isolation of a Bone-resorptive Factor from Human Cancer Ascites Fluid¹

Departments of Biochemistry [R. B. N., D. E. H., K. S.], Periodontology [R. B. N.], Microbiology [A. M. B., S. R. C.], and Oral Biology [W. S. L., H. W.], Boston University School of Medicine and Henry M. Goldman School of Graduate Dentistry, Boston University Medical Center, Boston, Massachusetts 02118

A protein fraction that induces the resorption of bone explants in organ culture was isolated from the ascitic fluid of patients with advanced cancer metastatic to the peritoneal cavity. Partial purification was achieved by means of gel filtration, affinity chromatography, and ion-exchange chromatography. The isolated fraction, the components of which have an apparent molecular weight of 60,000, was found to be heterogeneous by disc gel electrophoresis and to be composed primarily of proteins with relatively acidic electrophoretic properties. The specific bone-resorptive activity of this protein fraction was greatly increased over that of the unfractionated starting material, and the activity could be completely destroyed upon incubation with pronase and on heating. As determined by immunoassay and extraction procedures with various solvents, the bone-resorptive action of the isolated fraction was not attributable to the presence of parathyroid hormone, prostaglandin E₂ or vitamin D-like sterols. In parallel experiments the supernatants of phytohemagglutinin-stimulated normal human peripheral leukocytes were subjected to identical chromatographic techniques, and a protein fraction with a molecular weight of 60,000, which resembled the resorptive fraction isolated from cancer ascites fluid and which contained significant bone-resorptive activity, was also partially purified.

¹ This study was supported by USPHS Research Grants CA-22062, CA-14129, CA-12209, CA-15848, DE-03565, and GM-10374 and by an award from the American Cancer Society, IN-97B.

Received 9/29/77. Accepted 4/10/78.

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