Immunological identification and distribution of parathyroid hormone-like protein polypeptides in normal and malignant tissues.
Monoclonal and polyclonal antibodies recognizing human parathyroid hormone-like protein (PTHLP) have been produced using a series of recombinant and synthetic PTHLP peptides. These antibodies have been used to develop a two-site immunometric enzyme immunoassay which detects PTHLP[1-87] and PTHLP[1-141] but not PTH. The immunoassay detected PTHLP in extracts of squamous carcinomas and normal tissues at concentrations from 7-515 ng PTHLP[1-87]/mg protein. Immunoblotting of the extract which showed the highest immunoreactivity, a squamous carcinoma of the lung from a patient with hypercalcemia, revealed a major band having an apparent mol wt of 26,500 and several other higher mol wt bands. Similar polypeptides were observed by immunoblotting cell extracts from a cell line, SCaBER, which secretes immunoreactive PTHLP into its medium and also from tumors in nude mice derived from this cell line. Chaotropic agents did not alter the immunoblotting pattern, and antibodies to three different epitopes of PTHLP recognized these bands, indicating PTHLP expression in the extracts. Immunohistochemical staining of normal human tissue with these antibodies revealed several PTHLP-containing tissues and confirmed the results of the immunoassay, suggesting a paracrine role for PTHLP. Staining was observed in several neoplastic tissues including squamous cell carcinomas, lung carcinoma, bladder carcinoma, osteogenic sarcoma, and adenocarcinoma of the colon.
Kramer
S
S
Oncogene Science, Inc., Manhasset, New York 11030.
Reynolds
F H
FH
Castillo
M
M
Valenzuela
D M
DM
Thorikay
M
M
Sorvillo
J M
JM
D000230:Adenocarcinoma / Q000737:chemistry; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000918:Antibody Specificity; D002294:Carcinoma, Squamous Cell / Q000737:chemistry; D003110:Colonic Neoplasms / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D015151:Immunoblotting; D007150:Immunohistochemistry; D008175:Lung Neoplasms / Q000737:chemistry; D008297:Male; D051379:Mice; D008819:Mice, Nude; D008970:Molecular Weight; D012516:Osteosarcoma / Q000737:chemistry; D044162:Parathyroid Hormone-Related Protein; D011506:Proteins / Q000032:analysis* / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology; D014018:Tissue Distribution; D001749:Urinary Bladder Neoplasms / Q000737:chemistry
D000911:Antibodies, Monoclonal; C470121:PTHLH protein, human; D044162:Parathyroid Hormone-Related Protein; D011506:Proteins; D011994:Recombinant Proteins