Secretin/Vasoactive Intestinal Peptide-Stimulated Secretion of Bombesin/ Gastrin Releasing Peptide from Human Small Cell Carcinoma of the Lung1

ICANCER RESEARCH 46, 1214-1218, March 1986]

Secretin/Vasoactive Intestinal Peptide-stimulated Secretion of Bombesin/ Gastrin Releasing Peptide from Human Small Cell Carcinoma of the Lung1

Louis Y. Korman,2 Desmond N. Carney, Marc L. Citron, and Terry W. Moody

Medica/ Service (151W), Veterans Administration Medical Center, Washington, DC 20422 [L. Y.K., M.L.C.]; Department of Medicine and Biochemistry George Washington University School of Medicine, Washington, DC 20037 [T. W. M.Â¡;and National Cancer Institute-Navy Medical Oncology Branch National Cancer Institute and National Naval Medical Center, Bethesda, Maryland [D. N. C.Â¡

ABSTRACT autocrine factor for SCCL (12) growth. We studied the mecha nism of BLI secretion in several SCCL cell lines by examining Bombesin/gastrin releasing peptide-like immunoreactivity (BLI) the action of agents that increase intracellular cAMP. is found in the majority of small cell carcinoma of the lung (SCCL) Because of the results of these in vitro studies and the fact cell lines examined. Because BLI is present in high concentration that secretin stimulates hormone release in patients with gastrin in SCCL we studied the mechanism of BLI secretion from several producing tumors (Zollinger-Ellison syndrome), we examined the SCCL cell lines and in patients with SCCL. In cell line NCI-H345 action of i.v. secretin infusion on plasma BLI levels in several the structurally related polypeptide hormones secretin, vasoac- patients with SCCL, non-SCCL lung tumors, and patients with tive intestinal peptide, and peptide histidine isoleucine as well as theophylline, a phosphodiesterase inhibitor, N6,O2'-dibutyryl out any cancer. cyclic adenosine 3':5'-monophosphate, a cyclic nucleotide ana logue, increased BLI release by 16-120% and cyclic adenosine MATERIALS AND METHODS 3':5'-monophosphate by 36-350%. Similar results were ob Cell Culture. SCCL cell line NCI-H345 was established using a bone tained in SCCL cell line NCI-H209. i.v. injection of secretin (2 biopsy from a patient with SCCL who had relapsed following intensive units/kg) significantly increased plasma BLI in 2 patients with combination chemotherapy. In culture NCI-H345 is aneuploid, clonogenic extrapulmonary SCCL. These data suggest that SCCL cells in soft agarose, tumorigenic in nude mice, expresses elevated levels of 3,4-dihydroxy-L-phenylalanine decarboxylase (198 units/mg protein), possess receptors for secretin/vasoactive intestinal peptide and that receptor occupation stimulates in vitro and In vivo BLI neuron specific enolase (4075 ng/mg protein), and produces high quan tities of BLI (4.7 pmol/mg protein (2). The cell line was maintained secretion. continuously for 24 mo in a serum free defined HITES medium (13). Cells were incubated in 75-cm2 Falcon tissue culture flasks in a humidified INTRODUCTION atmosphere of 5% CO2/95% air at 37Â°C. One to 2 wk prior to each experiment stock cultures were passaged and then 3 days prior to use In the United States, small cell carcinoma of the lung accounts the cells were resuspended in fresh HITES medium. Other SCCL cell for 25,000-30,000 new cases of primary lung cancer each year lines were maintained as described previously (2). (1). Unlike the other major forms of lung cancer, in vivo and in BLI Radioimmunoassay. Rabbit antisera to bombesin was obtained vitro studies indicate that SCCL3 expresses several unique cy- as described previously (14). The antiserum recognizes bombesin and the structurally related gastrin releasing peptide equally well, because tochemical and functional characteristics including the presence they have the same COOH-terminal heptapeptide, and does not cross- of high levels of 3,4-dihydroxy-L-phenylalanine decarboxylase, react with secretin, VIP, or PHI (14). For the BLI determination cells or dense core neurosecretory granules, and the secretion of peptide supernatant was extracted with boiling 2 N acetic acid, centrifuged at hormones (2-4). These findings suggest that SCCL may origi 2,000 x g for 10 min to remove paniculate material, and the supernatant nate from endocrine (Kultschitzky) cells in the lung (5). However, was frozen, lyophilized, and stored at -70Â°C. The assay was performed recent studies indicate that SCCL may be derived from a he- by resuspending samples in 140 mw NaCI/16 mw NaPO4 (pH 7.4) matopoietic stem cell (6). Although several polypeptide hor containing 0.25% bovine serum albumin and incubating with rabbit anti- mones including adrenocorticotropic hormone, calcitonin, phys- bombesin serum at 1:100,000 dilution for 1 h at 4Â°C,then adding 5000 cpm of 125l-labeled tyrosine4 bombesin for a total vol of 400 pi and alaemin, arginine vasopressin, and neurotensin are produced by incubated for 16 h at 4Â°C.At the end of the incubation 200 pi of normal SCCL, BLI is frequently present (4, 7-9). Prior investigations rabbit serum at 1:200 dilution was added followed by 100 pi of goat anti- have demonstrated that hormone secretion from cultured endo rabbit serum at 1:20 dilution. After 30 min, 200 pi of 12% polyethylene crine cells including SCCL (10) may be mediated by the intracel- glycol was added to enhance precipitation and the samples were centri lular second messenger cAMP (11) and bombesin may be an fuged at 1000 x g for 20 min. The supernatant was removed and the pellet assayed for antibody-tracer complex in a LKB gamma counter Received 2/18/85; revised 7/30/85, 11/26/8S; accepted 12/2/85. 1This work was supported by funds from the Veterans Administration Research (LKB, Rockville, MD). Service and by National Cancer Institute grant CA-33767. Presented in part at the Venous blood samples were centrifuged in a Sorvall RC-5B refriger Annual Meeting of the American Federation for Clinical Research, Washington DC ated centrifuge at 500 x g for 10 min and the plasma was removed and April 1983. stored at -70Â°C. Prior to assay the plasma was thawed and processed 2 To whom requests for reprints should be addressed, at Gastroenterology Section (151 W), Veterans Administration Medical Center, 50 Irving St., NW, Wash using a modification of the method of Bohlen ef a/. (15). In brief, 2 ml of ington, DC 20422. plasma were added to 6 ml of 5% formic acid, 15% trifluoracetic acid, 3 The abbreviations used are: SCCL, small cell carcinoma of the lung; BLI, 9% HCI, and 1% NaCI. The precipitated plasma proteins were centrifuged bombesin/gastrin releasing peptide-like immunoreactivity; cAMP, cyclic AMP; VIP, at 2000 x g for 10 min and the supernatant was applied to a Sep-Pak vasoactive intestinal peptide; PHI, peptide histidine isoleucine; HITES (medium), RPMI 1640 medium supplemented with hydrocortisone (10 nM), bovine insulin (5 (Millipore). The peptides were eluted from the Sep-Pak using ethanol/0.2 fig/ml), transferrin (10 jig/ml), 17^-estradiol (10 nw), and sodium selenite (30 nM)- N HCI (1/1), and the eluate was frozen, lyophilized, and assayed for BLI cGMP, cyclic GMP. (14).

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Cyclic AMP Radioimmunoassay. Cyclic AMP was determined using mg protein by 5 min which declined to 39 pmol/mg protein by a minor modification (16) of the procedure described by Brooker ef al. 60 min. Similarly, 1 IÂ¿Msecretin, a gastrointestinal peptide with (17). At the times specified iced (4Â°C)absolute ethanol was added to an equal vol of either the cell suspension or the cell pellet, kept at 4Â°Cfor significant sequence homology to VIP (20), increased cAMP from 15 min and then frozen and stored at -70Â°C. Samples were thawed and 17 to 42 pmol/mg protein by 60 min. Without additions cAMP varied between 10 and 25 pmol/mg protein during the 60-min centrifuged at 10,000 x g for 5 min, and an aliquot of supernatant was incubation. Equivalent results were obtained for cell line NCI- removed and assayed for cAMP. Protein was measured by the method described by Lowry ef at. (18). H209 (data not shown). Secretin Infusion. Secretin (2 units/kg) was injected over a 30-s Fig. 2 shows that 1 Â»Msecretin increased BLI in the superna period into a large antecubital vein via a freely flowing i.v. At 15 min, just tant from 300 to 480 fmol/mg protein after 5 min. Secretion then prior to and at 2, 5,10, 30, and 60 min after secretin injection 5-ml blood remained constant in the presence of 1 /IM secretin. In contrast, samples were obtained, added to 0.5 ml of 20 HIM ethyleneglycol bis(/3- without additions supernatant BLI remained constant at 250 aminoethyl ether)-A/,A/,/v",A/'-tetraacetic acid and bacitracin (pH 7.4), 1 fmol/mg protein throughout the time course of the experiment. mg/ml and placed on ice. Symptoms, blood pressure, pulse, and respi These data indicate that 1 /.M secretin stimulated a rapid release ratory rate were monitored during and for 2 h after secretin injection and of BLI into the supernatant and this release coincided with the no untoward effects occurred. Informed consent, approved by the Med secretin-stimulated increase in cAMP. Similar results were ob ical Center Human Studies Subcommittee, was obtained from the pa tained using VIP. Because both the cAMP levels and secretion tients. Materials. RPM11640 with glutamine was obtained from Grand Island of BLI was maximal after 5 min this time point was used in all Biological, Grand Island, NY; VIP, PHI, secretin, bombesin, physalaemin, subsequent in vitro experiments. Table 1 shows that at 5 min, neurotensin, and cholecystokinin-octapeptide from Peninsula Laborato when secretagogue-stimulated cAMP was maximal, 1 ^M VIP, ries, Belmont, CA; bovine plasma albumin (Fraction V) from Armour PHI, or secretin significantly (21) increased BLI release by 25, Pharmaceutical, Phoenix, AZ; all other chemicals were from Sigma Chemical, St. Louis, MO. cGMP antiserum (preconjugated to a second antibody) and 125l-labeled 2'-O-succinyl(tyrosine methyl ester)cyclic GMP was from New England Nuclear, Boston, MA; 125l-labeled 2'-O-succi- SECRETIN lluM) 500 nylftyrosine methyl ester) cyclic AMP was from Melloy Laboratories, Springfield, VA; goat anti-cyclic AMP antibody was a gift from Dr. Gary Brooker, Georgetown University School of Medicine, Washington, DC; and the infused secretin was a gift from Dr. L. Gabel, Pharmacia Laboratories, Piscataway, NJ. 300 CONTROL

RESULTS ; 200 Previous studies suggest that SCCL cell lines of the classic morphology enriched in dense core neurosecretory granules 100 have high levels of BLI (7,19). To explore the response of SCCL cell lines to agents that increase cAMP, we examined the action 10 20 30 of 1 UM VIP and 1 UM secretin on cellular cAMP in SCCL cell TIME ii-.m lines NCI-H345 and H209. NCI-H345 and NCI-H209 are SCCL Fig. 2. Time course of secretin-stimulated BLI secretion in SCCL cell line NCI- cell lines which contain BLI. Cell line NCI-H345 was grown in H345. Results are mean Â±SE (bars) of BLI in the supernatant. Each value was HITES medium, whereas cell line NCI-H209 was grown in HITES determined in triplicate and the results are representative of 3 separate experi supplemented with 10% fetal bovine serum. Fig. 1 shows that in ments. cell line NCI-H345 1 MMVIP increased cAMP from 14 to 90 pmol/ Table 1 EHect of agents on cellular cyclic AMP and BLI release in SCCL cell line NCI-H345 90 For cAMP the results are the mean Â±SE of the number of experiments in parentheses and in each experiment each value was determined in duplicate. For 80 BLI release the results are the mean Â±SE of 5 separate experiments and in each experiment each value was determined in triplicate. Routinely, the cell pellet _70 C contained approximately 4.7 pmol BLI/mg protein in the absence of additions. 1 5 60 Without additions cellular cAMP was 37.9 pmol/mg protein and BLI in the super Ã›. natant was 260 fmol/mg protein. | 50 % of control

IÂ« Q. AdditionNoneVIP cAMP100 release100 Â±11(10) Â±10 Â¡30 125 Â±16" (1 â€žMl 462 Â±58(5f u 131 Â±12" 20 PHI (1,/M)Secretin 243 Â±19(5f (1 /*M) 136 Â±20(10)"111 115 Â±4C N,,02'-dibutyryl cAMP (1 mut) 141 Â±9" 10 Prostaglandin E, (10 JIM) Â±39(5) 111 Â±10 219 Â±30* 0 Theophylline+VIP (10 mw) 454 Â±79(5f 10 20 30 40 50 60 1180Â±152 (5)aBLI 244 Â±42* TIME iminl (1 pM)Cellular Fig. 1. Time course of VIP- and secretin-stimulated cAMP in SCCL cell line NCI- * P < 0.005 by Student's paired ( test (21). b P < 0.01 by Student's paired f test (21). H345. Results are expressed as mean Â±SE (bars) and each value was determined 0P < 0.05 by Student's paired f test (21). in duplicate. Results are representative of 3 separate experiments.

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31, and 15%, respectively, and that 1 ITIMN6,O2'-dibutyryl cAMP Because secretin increased cAMP and BLI secretion in vitro increased BLI release by 41 %. Neither 10 UM prostaglandin E1 and is used to stimulate gastrin release from gastrinomas in nor 1 mw nitroprusside (data not shown) significantly increased patients with Zollinger-Ellison syndrome (22), we examined the BLI release. VIP, PHI, and secretin increased cAMP levels by time course of secretin-stimulated BLI secretion in several pa 360, 140, and 40%, respectively, whereas prostaglandin EÃ•had tients with SCCL including two with extensive extrapulmonary no effect. Theophylline (10 HIM) alone caused a 119% increase disease. As described previously, basal levels of plasma BLI in BLI release and a 350% increase in cellular cAMP. Adding 1 were significantly greater in the patients with extrapulmonary UMVIP slightly increased the theophylline-stimulated BLI release SCCL than in the patients with non-SCCL tumors or normal but dramatically augmented the theophylline-stimulated cellular controls (23). In the two patients with extensive SCCL the cAMP (Table 1). Since the agents tested might also increase BLI injection of secretin (2 units/kg) rapidly increased plasma BLI release by increasing cGMP we measured cellular cGMP under from 60 to 210 PM by 2 min and 91 pw by 1 h, respectively (Fig. the same conditions as described in Table 1 and in the presence 3). In one patient with non-SCCL limited to the thorax, secretin of 1 rriM nitroprusside. Only 10 mw theophylline and 1 mw stimulated a transient increase in plasma BLI. The reason for nitroprusside increased cellular cGMP. Furthermore, 1 UM bom- this small response is not clear. In contrast, in three untreated besin, 1 MMcholecystokinin, 1 MMphysalaemin, 1 MMneurotensin, patients with SCCL limited to the thorax (Fig. 3A) and in two 1 MMcalcitonin, 10 MMphenylephrine, and 10 MMcarbamylcholine patients without any evidence of tumor (data not shown) secretin did not increase either cGMP or cAMP. infusion did not alter plasma BLI. Furthermore, after chemother To determine whether the secretagogue-stimulated increase apy-induced remission in the patient with extrathoracic SCCL in cAMP and BLI secretion was unique to cell line NCI-H345, we basal plasma BLI was reduced by approximately 50% (36 versus examined the action of VIP, secretin and theophylline on another 74 pw) and there was no increase in secretin-stimulated plasma SCCL cell line (Table 2). In NCI-H209,1 MMVIP and 1 MMsecretin BLI (Fig. 38). increased cAMP from 5.3 to 19.6 and 9.9 pmol/mg protein, respectively, and BLI secretion from 567 to 699 and 787 fmol/ mg protein, respectively. Although the theophylline-stimulated DISCUSSION increase in cAMP was equal to that of VIP, the theophylline- VIP and secretin are widely distributed peptide hormones stimulated increase in BLI was significantly greater, 1134 versus which mediate a variety of physiological responses including 699 fmol/mg protein, respectively. gastrointestinal secretion, relaxation of gastrointestinal, vascular, and respiratory smooth muscle, lipolysis in adipocytes, glyco- Table 2 genolysis in hepatocytes, pituitary hormone secretion, and excit Action of agents on cellular cAMP and BLI secretion in cell line NCI-H209 ation and hyperthermia after injection into the central nervous For cAMP and BLI the results are the mean Â±SE of 2 separate incubations and system (21, 24). VIP receptors, some of which bind VIP with are representative of 3 separate experiments. For cAMP each value was determined in duplicate and for BLI each value was determined in triplicate. Routinely the cell high affinity and secretin with low affinity (25, 26), and secretin pellet contained 18.3 pmol BLI/mg protein. receptors are coupled to adenylate cyclase in normal and malig cAMP secretion nant intestinal epithelium, pancreas, gastrointestinal and cardio AdditionsNoneVIP (pmol/mgprotein)5.3 (fmol/mgprotein)567 vascular smooth muscle, adipocytes, hepatocytes, a pituitary Â±0.3 Â±92 tumor cell line, brain, and leukocytes (21,24,27,28). The present 19.6 Â±2.0a 699 Â±59Â° (1UM)Secretin 9.9 Â±1.5a 787 Â±66" studies suggest that VIP and secretin receptors are also present (1 UM) 17.4 Â±0.9*BLI 1134 Â±154" Theophylline (10 mM)Cellular on several SCCL cell lines and that occupation of these receptors " P < 0.01 by Student's paired f test (21). increases adenylate cyclase activity and stimulates BLI release " P < 0.05 by Student's paired ( test (21). via a cAMP-mediated pathway. Furthermore, other agents that

A. X SECRETIN|2U/kg) B. SECRETIN (2U/kg)

200 500

Fig. 3. A, secretin-stimulated plasma BLI in patients with SCCL and non-SCCL tumors. Se cretin infusion and plasma BLI determinations 160 400 were performed as described in "Materials and Methods." Patient M. G. (â€¢),treated, recurrent, extrapulmonary SCCL; Patient J. P. (O), H120 treated, metastatic. well-differentiated adeno- I 300 carcmoma; Patients L. R. (A), F. P. (T), and E. R. (â€¢)untreated SCCL limited to the thorax. B, secretin-stimulated plasma BLI in a patient with 80 extrathoracic SCCL. Secretin infusion and 200 plasma BLI determinations were performed as described in "Materials and Methods." Patient

W. T. pretreatment (â€¢)and after 4 courses of 40 chemotherapy (A.) resulted in a 90% reduction 100 in extrathoracic tumor burden.

-15 30 60 -15 30 60 TIME {mini TIME (min)

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Downloaded from cancerres.aacrjournals.org on October 1, 2021. © 1986 American Association for Cancer Research. BOMBESIN/GASTRIN RELEASING PEPTIDE SECRETION FROM HUMAN SCCL increase cAMP by inhibiting phosphodiesterase activity or by so that tumor BLI release may not be detectable, and (c) the simulating an increase in intracellular cAMP also increase BLI absence of BLI in the tumor cells. Secretin-stimulated BLI release release (10). Thus, SCCL cells, like other cultured endocrine is unlikely to be of use in the differential diagnosis of disease cells, possess an intact cAMP-mediated pathway for stimulating limited to the thorax. However, the presence of a positive re hormone release. Because binding studies were not performed sponse to secretin may be useful in predicting response to the present results do not distinguish between two separate therapy, diagnosing extrathoracic spread, or documenting recur classes of receptors for VIP and secretin or one class of recep rence. Clearly, larger numbers of normal patients and patients tors for VIP/secretin which recognizes each with different affini with limited and extensive non-SCCL tumors need to be studied ties (25, 26); nor do these studies define the pharmacological to determine the clinical utility of this provocative test. Another characteristics of the VIP/secretin receptor. potential clinical application for these findings is in the treatment That secretion from SCCL cell lines is also calcium dependent of SCCL. Recent observations suggest that changes in cellular is suggested by prior studies (10, 29) as is the finding that cAMP alter the growth rate and metabolism of SCCL and other theophylline-stimulated BLI release is significantly greater than cells in vitro (38, 39). Since BLI acts as an autocrine growth VIP-stimulated BLI release, whereas there is no difference be factor for SCCL, regulation of BLI release from SCCL may play tween VIP- and theophylline-stimulated cellular cAMP (Table 1). a role in regulating tumor growth (12). Previous studies indicate that carbamylcholine and methylxan- thine phosphodiesterase inhibitors increase pancreatic enzyme ACKNOWLEDGMENTS secretion by modifying cellular calcium fluxes (16). In addition, carbamylcholine stimulates BLI release (10) but does not in We thank Drs. L. Recant, A. Gazdar, J. Minna, M. Cohen, and S. Krasnow for their support and suggestions and V. Bertness and G. Lebovic for technical crease cAMP or cGMP production. Thus, BLI may be secreted assistance. We thank Paula Kittrell and Jan Parker for preparing this manuscript from SCCL cells by altering the intracellular cAMP levels or by for publication. altering calcium flux. In classic SCCL cell lines one of the characteristic findings is the presence of BLI. Of the polypeptide hormones examined REFERENCES prior work indicates that 94% (32 of 34) of the SCCL cell lines 1. Rosenow, E.G.. and Carr, D. T. Bronchogenic carcinoma. Ca-a Cancer Journal and none (0 of 13) of a variety of non-SCCL cell lines contained for Clinicians, 29: 233-246, 1979. 2. Carney, D. N., Broder, L., Edelstein, M., Gazdar, A. F., Hasen, M., Haveman, BLI (19). In contrast to cultured cells, extracts of 17% (15 of 88) K., Mattews, M. J., Sorenson, G. D., and Videlov, L. Experimental studies of of non-SCCL primary lung tumors (squamous, adenocarcinoma, the biology of human small cell lung cancer. Cancer Treat. Rep., 67: 27-35, and large cell carcinoma) contained BLI (30). BLI is also found in 1983. 3. Pearse, A. G. E. The cytochemistry and ultrastructure of polypeptide hormone moderate densities in the brain (14, 31), carcinoid tumors (32), producing cells of the APUD series. J. Histochem. Cytochem., 77: 303-313. gastrointestinal tract (33), and fetal but not adult lung (34). 1969. 4. Baylin, S. B., and Mendelsohn, G. Ectopie (inappropriate) hormone production Although plasma levels of BLI are occasionally elevated in pa by tumors. Endocr. Rev.. 7: 45-77, 1980. tients with extensive SCCL (highest detected level is 2 nw), with 5. Bonikos, D. S.. and Bensch, K. G. Endocrine cells of bronchial and bronchiolar limited disease plasma levels are similar to those in normal epithelium. Am. J. Med., 63: 765-771, 1977. 6. Ruff, M. R., and Pert, C. B. Small cell carcinoma of the lung: macrophage- controls. It is unlikely therefore that routine screening of blood specific antigens suggest hematopoietic stem cell origin. Science (Wash. DC)., for BLI will be useful in early detection or follow-up of patients 225: 1034-1037, 1984. with SCCL. Our in vitro findings suggested that a provocative 7. Moody, T. W.. Pert, C. B.. Gazdar, A. F., Carney, D. N., and Minna, J. D. High levels of intracellular bombesin characterize human small cell line carcinoma. test using secretin to stimulate BLI release might be useful in Science (Wash. DC)., 274. 1246-1248, 1981. the evaluation of SCCL. In patients with insulinoma and gastri- 8. Wood, S. M., Wood, J. R., Ghatei, M. A., O'Shaugnessy, D., and Bloom, S. R. Bombesin, somatostatin and neurotensin-like immunoactivity in bronchial noma, glucagon and secretin infusion increase serum insulin and carcinoma. J. Clin. Endocrinol. Metab., 53: 1310-1312,1981. gastrin concentration (22, 34), respectively. In vitro studies indi 9. Moody, T. W. Bombesin-like peptides in the normal and malignant lung. In: K. cate that glucagon and secretin act directly on tumor cells to L. Becker and A. F. Gazdar (eds.). The Endocrine Lung in Health and Disease, pp. 328-355. Philadelphia: W. B. Saunders, 1984. increase insulin and gastrin release (36, 37). The finding that 10. Sorenson, G. D., Pettengill, O. S., Cate, C. C., Ghatei, M. A., and Bloom, S. secretin increased plasma BLI in the two patients with metastatic R. Modulation of bombesin and calci ton m secretion in cultures of small cell carcinoma of the lung. In: K. L. Becker and A. F. Gazdar (eds.). The Endocrine SCCL supports our in vitro findings that SCCL possesses func Lung in Health and Disease, pp. 596-601. Philadelphia: W. B. Saunders Co., tionally intact secretin receptors. 1984. That the basal and secretin-stimulated increase in plasma BLI 11. Vale, W., Rivier, C., Yang, L., Minick, S., and Guillemin, R. Effects of purified hypothalamic corticotropin-releasing factor and other substances on the se was due to BLI release from SCCL cells and not from another cretion of adrenocorticotropin and beta-endorphin-like immunoactivities in vitro. source of BLI is suggested by the following observations: (a) in Endocrinology, 703: 1910-1915, 1978. two patients with extrathoracic SCCL basal plasma BLI was 12. Cuttitta, F., Carney, D. N., Mulshine, J., Moody, T. W., Fedorko, J., Fischler, A., and Minna, J. D. Bombesin-like peptides can function as autocrine growth approximately 60 and 74 pM, whereas basal levels of BLI are 10 factors in human small cell lung cancer. Nature (Lond.), 376: 823-827, 1985. to 50 pM in normal patients and patients with limited SCCL; (b) 13. Carney, D. N., Bunn, P. A., Gazdar, A. F., Pagan, J. A., and Minna, J. D. Selective growth of small cell carcinoma of the lung obtained from patient secretin did not increase plasma BLI from the three patients with biopsies in serum-free hormone supplemented medium. Proc. Nati. Acad. Sci. SCCL limited to the thorax or from two patients without tumors. USA, 78. 3185-3189, 1980. Furthermore, significant reduction in tumor burden with chemo 14. Moody, T. W., and Pert, C. B. Bombesin-like peptides in rat brain. Biochem. therapy abolished the secretin-stimulated increase in plasma BLI. Biophys. Res. Commun., 90: 7-14, 1979. 15. Bohlen, P., Castillo, F., Shibasaki, T., Ling. N., Guillemin, R. New micrometh- The absence of secretin-stimulated increase in plasma BLI in odology for the isolation of peptides. In: Peptides: Structure and Biological patients with limited SCCL may be due to (a) rapid degradation Function, pp. 109-112. Rockford, IL: Pierce Chemical Co., 1979. 16. Korman, L. Y., Walker, M. D., and Gardner, J. D. Action of theophylline on of BLI in the lung or serum after secretin-stimulated release, (b) secretagogue-stimulated amylase release from dispersed pancreatic acini. Am. the release of small amounts of BLI relative to the plasma levels J. Physiol., 239: G324-G333, 1980.

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Downloaded from cancerres.aacrjournals.org on October 1, 2021. © 1986 American Association for Cancer Research. Secretin/Vasoactive Intestinal Peptide-stimulated Secretion of Bombesin/Gastrin Releasing Peptide from Human Small Cell Carcinoma of the Lung

Louis Y. Korman, Desmond N. Carney, Marc L. Citron, et al.

Cancer Res 1986;46:1214-1218.

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