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A and CCKB/Gastrin Receptors in Human Tumors

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A and CCKB/Gastrin Receptors in Human Tumors (CANCER RESEARCH57, 1377-1386, April 1. 19971 Cholecystokinin(CCK)-A and CCKB/Gastrin Receptors in Human Tumors Jean Claude Reubi,' Jean-Claude Schaer, and Beatrice Waser Division of Cell Biology and Experimental Cancer Research, institute of Pathology, University of Berne, Murtenstrasse 31. CH-3010 Berne, Switzerland ABSTRACT The gastrointestinal peptides gastrin and/or CCK have been impli cated in various regulatory functions; as neurotransmitters in the Cholecystokinin (CCK)-A and CCK.B/gastrln receptors were evalu brain; and in the regulation of various functions of the gastrointestinal ated with in vitro receptor autoradlography in 406 human tumors of tract, primarily at the level of the stomach, pancreas, and gallbladder various origins using a sulfated lasI@labelod CCK decapeptide analogue (8). In addition, they can act as physiological growth factors in most lasI.([email protected], Me@1).CCK 26-33 and *asI.Inbelod Leu15-gastrin as radioligands. CCK-B/gastrln receptors were found frequently in medul parts of the gastrointestinal tract (9—11) and also as stimulatory lat@r thyroid carcinomas (92%), In small cell lung cancers (57%), in growth factors in several neoplasms, such as colonic and gastric astrocytomas (65%), and in stromal ovarian cancers (100%). They were cancers (12—14).Gastrin and CCK possess the same terminal five found occasionally In gastroenteropancreatic tumors, breast, endometrial, amino acids at their COOH terminus, which is the biologically active and ovarian adenocarcinomas. They were either not expressed or rarely site; their actions are mediated by two different receptor types, expressed In colorectal cancers, differentiated thyroid cancers, non-small CCK-Aand CCK-Breceptors(15, 16), which can be distinguished cell lung cancers, meninglomas, neuroblastomas, schwannomas, glioblas pharmacologically by their low (CCK-A) versus high (CCK-B) affin tomas, lymphomas, renal cell cancers, prostate carcinomas, and the re ity for gastrin. maining neuroendocrine tumors (Le., pituitary adenomas, pheochromo CCK-Aand CCK-B/gastrinreceptorshave been identifiedin sev cytomas, paragangilomas, and parathyroid adenomas). CCK-A receptors eral normal tissues; CCK-B/gastrin receptors are present in the gut were expressed rarely In tumors except In gastroenteropancreatic tumors (38%), meningiomas (30%), and some neuroblastomas (19%). The iden mucosa and in the brain (8, 17, 18). CCK-A receptors are present in tified CCK-A and CCK-B receptors were specific and of high affinity In the gallbladder, pancreas, and brain (8, 19). The presence of receptors the subnanomolar range. The rank order of potency of various CCK for gastrin and CCK in tumors has also been reported. It is well analogues was: sulfated CCK-8 L-364,718 )D nonsulfated CCK-8 L. established that small cell lung cancers often express CCK-B/gastrin 365,260 gastrIn for CCK-A receptors and sulfated CCK-8 > gas receptors, whereas non-small cell lung cancers do not express them trin nonsulfated CCK-8 > L-365,260 > L-364,718 for CCK-B recep (20, 21). However, the findings are more equivocal for gastrointestinal tors. CCK-B receptors could also be selectively and specifically labeled cancers. Whereas earlier studies have reported CCK-B/gastrin recep with a newly designed nonsulfated lasI.(D.Tyr@Gly,Nle@')-CCK 26-33. tors in colon cancers and gastric cancers (22), more recent investiga Gastrin mRNA measured by in sins hybrkllzation was present In most tions failed to find high-affinity CCK-B/gastrin receptors in most of CCK-B receptor-positive small cell lung cancers, breast tumors, and ovarian tumors, representIng the molecular basis of a possible autocrine these tumors (23). Recently, an unexpected high incidence of CCK-B growth regulation of these tumors. Gastrin and CCK mRNAs were lack receptors was identified in medullary thyroid carcinomas, whereas mg In medullary thyroid cancers. Thus, these results may have patho differentiated thyroid cancers were not expressing them (24). Little genie, diagnostic, differential diagnostic, and therapeutic implications. information is available, however, about the CCK-B/gastrin receptor incidence in other human tumors. INTRODUCTION The main goal of the present study was to investigate the CCK-B/ gastrmnand CCK-A receptor prevalence in a variety of human tumors, There has been recently an increasing interest in the expression of including a large group of neuroendocrine tumors. Receptors can be peptide receptors by human tumors. The in vitro identification of these best evaluated in complex human tissues with receptor autoradiogra receptors in tumors was indeed found to be an important prerequisite phy, which preserves the morphological integrity of the tissues and for the evaluation of new potential clinical applications of the corre allows the precise localization of the receptors. Adequate ligands have sponding peptides (1—3). For example, the discovery of the high been shown previously to identify CCK-B and/or CCK-A receptors in incidence of somatostatin receptors in several types of human cancers animal and human tissues (17, 24—26).Therefore, the following two has led to the development of in vivo somatostatin receptor imaging of ligands were used routinely in all cases: ‘251-Leu'5-gastrin, which tumors, allowing the in vivo localization in patients of somatostatin directly identifies gastrin receptors, and ‘251-CCKdecapeptide, which receptor-positive tumors after injection of a labeled somatostatin identifies CCK-B/gastrin or CCK-A receptors based on whether the analogue (4, 5). The same may be true for VIP@receptors as well (6). ligand is displaced by nanomolar concentrations of CCK and gastrin The presenceof peptidereceptorsin tumorsmayalso be of therapeu or of CCK only. tic interest. Tumoral somatostatin receptors were shown to mediate the successful symptomatic treatment of neuroendocrine tumors with MATERIALS AND METHODS somatostatin analogues (2), and the growth of VIP receptor-positive breast cancers can be inhibited by a synthetic VIP receptor antagonist Aliquots of surgically resected tumors or of biopsies submitted for diag (7). These examples suggest that the evaluation of the expression of nostic histopathology were frozen immediately after surgical resection and other peptide receptors in tumors should be of considerable potential stored at —70°C.Thespecimens originated from several different clinical interest. institutions, and some have been used previously for other purposes. The following tumors were investigated: neuroendocrine tumors, including small cell lung cancers, pituitary adenomas, endocrine gastroenteropancreatic tu Received 10/15/96; accepted 1/31/97. mors, medullary thyroid carcinomas, parathyroid adenomas, pheochromocy The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with tomas, paragangliomas, and neuroblastomas; non-small cell lung cancers; 18 U.S.C. Section 1734 solely to indicate this fact. colorectal adenocarcinomas; papillary and follicular thyroid carcinomas (dif I To whom requestsfor reprintsshouldbe addressed.Phone:031-63232 42; Fax: ferentiated thyroid cancers); breast carcinomas; astrocytomas; glioblastomas; 031-632 89 99 or 632 49 95. schwannomas; meningiomas; endometrial and prostate carcinomas; ovarian 2 The abbreviations used are: VIP, vasoactive intestinal peptide; CCK, cholecystoki nin; @‘I-gastrin,‘251-Leu'5-gastrinI; @‘l-CCK,‘251-D-Tyr-Gly-Asp-Tyr(SO3H)-Nle cancers (including adenocarcinomas and stromal tumors); renal cell tumors; Gly-Trp-Nle-Asp-Phe-amide, i.e., @‘I-(i-Tyr-Gly,Nle28'31)-CCK26-33. and non-Hodgkin's lymphomas (Table I). 1377 Downloaded from cancerres.aacrjournals.org on September 24, 2021. © 1997 American Association for Cancer Research. CCK-A AND CCK-B/GASTRIN RECEPTORS IN HUMAN TUMORS tumorsTumorTable 1 CCX receptor incidence in were preincubated as described above and then incubated at room temperature receptorsbNeuroendocrinetypeCCK-B receptors―CCK-A for 150 mm in a solution containing the same medium as for the preincubation tumorsMedullary except that BSA was omitted, and the following compounds were added: 45 @M (8%)cSmall thyroid carcinomas22 of 24 (92%)c2 of 24 ‘@I-CCK(2000Ci/mmol; Anawa), 0.025% bacitracin,1 mMDTI', 2 jxg/ml (0%)Gastroenteropancreaticcell lung cancers8 of 14 (57%)0 of 14 chymostatin, and 4 @xg/mlleupeptin, pH 6.5. The slides were then treated as (38%)Growth tumors7 of 32 (22%)12 of 32 described above for ‘25I-gastrinreceptorautoradiography. 9(0%)Inactivehormone pituitary adenomas0 of 9 (0%)0 of 10(0%)Pheochromocytomas0pituitary adenomas0 of 10 (0%)0 of The autoradiograms were quantified using a computer-assisted image-pro 10(0%)Paragangliomas0 of 10(0%)0 of cessing system, as described previously (27, 28). Tissue standards for iodinated 10(0%)Neuroblastomas1 of 10 (0%)0 of compounds (Amersham UK, Little Chalfont, United Kingdom) were used for 16(19%)Parathyroid of 16 (6%)3 of 4(0%)Tumors adenomas0 of 4 (0%)0 of this purpose. A tissue was defined as receptor positive when the absorbance systemAstrocytomasIof the nervous measured over a tissue area in the total binding section was at least twice the (0%)Meningiomas1 1of 17(65%)0 of 17 absorbance of the nonspecific binding section. (30%)Schwannomas0 of 27 (4%)8 of 27 Receptor Autoradlography UsIng Nonsulfated tasI@(D@Tyr@Gly, (0%)Glioblastomas0 of I3 (0%)0 of 13 10(0%)Tumors of 10(0%)0 of Me@')-CCK 26—33.Nonsulfated (D-Tyr-Gly, Nle28'31)-CCK26—33was
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