Morphine Cross-Reacts with Somatostatin Receptor SSTR2 in the T47D Human Breast Cancer Cell Line and Decreases Cell Growth'

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ICANCERRESEARCH55. 5632-5636. December 1, 19951 Morphine Cross-Reacts with Somatostatin Receptor SSTR2 in the T47D Human Breast Cancer Cell Line and Decreases Cell Growth'

Anastassia Hatzoglou,2 L'Houcine Ouafik, Efstathia Bakogeorgou, Kyriaki Thermos, and Elias Castanas

Laboratories of Experimental Endocrinology (A. H., E. B., E. C] and Phannacology (K. TI, University of Crete, School of Medicine, and University Hospital, P.O. Box 1393, Heraklion GR-71 I 10, Crete, Greece, and Laboratoire de Cancerologie Ezperimentale, CJF INSERM 93-1 1 (A. H.. E. C.], and Neuroendocrinologie Experimentale, U 297 INSERM IL 0.1, FacultÃ©de Medecine Nord, Marseille, France

ABSTRACT act with the SSTR system, providing a possible explanation for their observed inhibitory effect on cell growth. In a previous study, we found that morphine decreases, in a dose dependent manner, the cell growth of T47D human breast cancer cells, despite the lack of p opioid receptors and an interaction of morphine with MATERIALS AND METHODS other opioid sites. We have therefore examined a possible interaction of Cell Cultures. The human breast cancer cell line T47D (originally isolated morphine with other membrane receptor systems of the cell. The present from a pleural effusion of breast adenocarcinorna) was obtained at passage 90. study describes for the first time an interaction between p-acting opioid Cells were routinely grown in RPMI, supplemented with 10%heat-inactivated drugs and the somatostatinergic system. We have found that [tasI]Tyr1t@ fetal bovine serum. They were cultured at 37Â°Cina humidified atmosphere of somatostatin binds with high affinity to T47D cells. Analysis ofthe binding 5% CO2 in air. data showed the presence of two components: one with hinjaaffinity but Somatostatin Binding Conditions. About 106 cells/well in monolayer low capacity (Kd,0.145 viM;1450 sites/cell), and another of lower affinity were used for saturation and displacement binding experiments. Before bind but higher capacity (Kd,l.l92 nM; 11920 sites/cell). Somatostatin-14 and ing, cells were washed twice with 2 ml of PBS. Binding was performed in PBS somatostatin-28 showed multiphasic displacement curves, indicating het in a total volume of 0.5 ml, containing 2]l somatostatin, without (total erogeneity of binding sites. The latter was confirmed by reverse transcrip binding) or with (nonspecific binding) a 1000-fold molar excess of somatosta tion-PCR, which revealed the existence of the somatostatin receptor sub tin-14. For saturation binding, at least 8â€”10pointswith different concentra types 2 and 3 (SSTR2 and SSTR3), with a relative mRNA concentration ions of radiolabeled peptide were performed in duplicate. The cells were of 85 and 15%, respectively. Morphine and the morphinomimetic peptide incubated for 2 h at room temperature (18â€”22Â°C).Atthe end of the incubation morphiceptine (Tyr-Pro-Phe-Pro-NH2) displace somatostatin from its period, the unbound radioactivity was eliminated by washing the cells twice binding sites. Further analysis indicated that p-acting opioids interact with 2 ml cold PBS. Cells were removed from plates with 0.4 ml 2 N NaOH, with the SSTR2 receptor subtype. and the bound radioactivity was counted in a -y counter (Tncarb Series; Packard), with a 95% efficiency for 1251,Bindingwas repeated at least three times, and the results were analyzed by the Origin Version 3.5 package INTRODUCTION (MicroCal Co.) using equations described by Munson and Rodbard (24). Cell Growth Conditions. Cells wereplatedin 24-well ELISAplatesat an The somatostatinergic and opioid systems are inhibitory and are initial density of25 X l0@cells/wellsupplemented with 1ml medium/well. All involved in the decrease of hormone and neurotransmitter secretion drugs were added to cultures 1 day after seeding (designated as day 0) to (1â€”6).Furthermore, they have been implicated in the control of tumor ensure uniform attachment of cells at the onset of the experiments. Cells were growth in different organs, including breast (1, 4, 7â€”13).Indeed, grown for a total of4 days, with daily change of the medium containing opioid somatostatin and opioid immunoreactivities have been detected in drugs or somatostatin analogues. All added drugs were dissolved shortly before tumor, but not in normal breast cells (3, 14, 15), and specific receptors use. Cell Proliferation. Cell growthwas measuredbythetetrazoliumsaltassay have been characterized in primary human breast tumors and tumor (25). Cells were incubatedfor 4 h at 37Â°Cwiththe tetrazoliurnsalt [3-(4,5- cell lines (4, 12, 13, 16â€”23),producing a dose-dependent inhibition of dimethylthiazol cell proliferation. 2-yl)-2,5-diphenyl tetrazolium bromide], and metabolically active cells re @ Recently, we have characterized and K opioid receptors in the duced the dye to purple formazan. Dark blue crystals were dissolved with T47D cell line and have shown that different opioids inhibit cell propanol. The absorbance was measured at 570 nm and compared against a @ proliferation (22). No opioid binding sites were found, however, standard curve of known numbers of T47D cells. All experiments were whereas morphine (the prototype @.tligand) shows a dose-dependent performed a minimum of three times, in triplicate. inhibitory effect on the growth of cells, which is not reversed by the Detection of SSTR mRNA by RT-PCR. Total RNA was preparedfrom opioid antagonist diprenorphine. Morphine does not cross-react with the T47D cell line using the guanidinium thiocyanate/phenol/chloroform pro cedure (26). Total RNA (5 @xg)fromT47D cells was reverse transcribed into any other opioid receptor subtype (@ or K). Our results suggest a cDNA using 1 @xgoligo(dT)12_18asprimer in 20-pi reaction volume [50 m@i nonopioid-mediated action of morphine and morphiceptine. Because Tris (pH 8.0)-75 mM KC1-5 mM MgC12-5 @xMDIT-SO @xg/mlBSA-20 units of somatostatin and opioid receptors belong to the same membrane RNasin-0.5 mM each of four dNTPs-400 units of Moloney murine leukemia protein superfamily (5, 6), we have investigated a possible interaction virus reverse transcriptase) at 37Â°C for 90 mm. A negative control was of morphine with the somatostatinergic system. perfonned with the first strand synthesis reaction and where the RNA sample In the present work, we have characterized the SSTRs3 in the T47D was omitted. RT mixtures without reverse transcriptase were included as cell line, both pharmacologically and by RT-PCR. We further report controls for each sample to detect any genomic DNA contamination. After the that morphine and the morphinomimetic peptide morphiceptine inter incubation, cDNA derived from 500 ng total RNA was amplified in a 50-pJ reaction containing 50 mMKC1,10 mMTris (pH 8.3), 1.5 mMMgCl2, 10 nmi Dli', 1 mMof each oligonucleotide, and 2.5 units of Taq DNA polymerase. Received 5/2/95; accepted 8/31/95. Optimal temperatures and cycling conditions were established for all five 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 SSTRs, as follows: SSTR1, denaturation at 95Â°CforS mm, and then 35 cycles 18 U.S.C. Section 1734 solely to indicate this fact. at 95Â°Cfor60 s, 65Â°Cfor90 s, and 72Â°Cfor 120 s [sense, 5'-AAA-TGC I This work was partially supported by the University of Crete Research Committee, GTC-CCA-GAA-CGG-GAC-CT-(C)-3'; antisense, 5'-CAG-GTf-CTC Ministry of Health (KESY), and Hellenic Anticancer Society grants. AGQ-TTG-GAA-GTC-TT-(C)-3']; SSTR2, denaturation at 95Â°C for S mm, 2 To whom requests for reprints should be addressed. 3 The abbreviations used are: 55Th, somatostatin receptor; RT-PCR, reverse transerip and then 35 cycles at 95Â°Cfor60 s, 60Â°Cfor90 s, and 72Â°Cfor120 s (sense, tion-PCR; BIM 23034C. o-Phe-c[Cys-Tyr-o-Trp-Lys-Val-Cys]Nal-NH2. 5'-CAT-ATA-GGA-TAG-GQT-TGG-CAC-AGC-TGT-T-3'; antisense, 5'- 5632

TAC-ATC-CTC-AAC-CTG-GCC-ATC-GCA-GAT-GA-3'); SSTR3, denatur the presence of multiple somatostatin binding sites in T47D breast ation at 95Â°C for 5 mm, and then 40 cycles at 95Â°C for 60 s and 67Â°C for 4 mm cancer cells. (sense, 5'-AGA-ACG-CCC-TGT-CCT-ACT-GGC-C-3'; antisense, 5'-TGA The specificity of the [â€˜251]Tyr'â€˜somatostatin binding to SSTRs AGC-GGT-AGG-AGA-GGA-AGC-C-3'); SSTR4, denaturation at 95Â°CforS is shown in Fig. 2A. Displacement of [â€˜251}Tyr'â€˜-somatostatin mm, and then 35 cycles at 95Â°Cfor60 s, 60Â°Cfor90 s, and 72Â°Cfor120 s from its receptor sites by somatostatin-14 and somatostatin-28, at (sense, 5'-ATG-GTC-GCT-ATC-CAG-TGC-3'; antisense, 5'-GGG-CTC CTC-AGA-AGG-TGG-T-3'); and SSTR5, denaturation at 95Â°CforS mm, and concentrations ranging from l0 â€˜â€˜to lO_6 M, produced parallel then 40 cycles at 95Â°C for 40 s, 65Â°C for 30 s, and 72Â°C for 90 s (sense, displacement curves, indicating an interaction with the same re 5'-CTC-TFG-GTG-TFC-GCG-GAC-GT-3'; antisense, 5'-CAG-GTF-GAC ceptor populations. Both curves were multiphasic, suggesting, as GAT-GTF-GAC-GGT-GAA-G-3'). did the saturation data, the presence of multiple somatostatin Southern Blots. Twenty pA of the PCR product were fractionatedon binding sites. No displacement was found by corticotrophin-releas agarose gels in TBE buffer (89 mMTris, 2 mMEDTA). After staining with ing hormone. On the contrary, the displacement of [â€˜25I]Tyr'â€˜- ethidium bromide, the gels were photographed, and then treated and trans somatostatin-14 by BIM 23034C, a selective SSTR2 analogue, ferred by capillarity to a Hybond-N membrane (Amersham, Amersham, United produced a monophasic displacement curve (Fig. 2B) and depicted Kingdom). The specificity of the PCR product was checked by Southern blot hybridization by using an internal sequence as probe for all SSTRs separately. an IC50 of 5 nM. Comparing Figs. 2A and 2B, it appeared that the Radiochemicals and Chemicals. [â€˜251 -somatostatin (2000 Cilmmol) first site detected in the somatostatin-l4 and somatostatin-28 dis was obtained from Amersham. Morphine was a gift from Farmacopia, and placement curves, and with IC50s of 0.8 and 0.9 nM, respectively, diprenorphine was from Reckit and Coleman Co. Cell culture media were from is the SSTR2 subtype. This was further confirmed by displacement GIBCO. The sornatostatinpeptides were obtained from Bissendorff Biochemi of [â€˜ â€˜-somatostatin by somatostatin- 14 and somatosta cals (Hanover, Germany). Sandostatin was a gift from Sandoz (Basel, Swit tin-28 in the presence of BIM 23034C (not shown). These data are zerland). All other peptides were from Sigma Chemical Co. (St. Louis, MO). summarized in Table 1. All chemicals were from Merck (Darmstad, Germany). Molecular Biology products were from Boehringer Mannheim (Mannheim, Germany). Characterization of SSTR Subtypes by RT-PCR. To analyze the heterogeneity of the binding sites, we have investigated further the identity of the SSTR subtypes present in T47D cells by performing an RESULTS analysis of their mRNA by RT-PCR. As shown in Fig. 3, in these Identification and Characterization of SSTRS on T47D Breast cells, only SSTR2 and SSTR3 mRNA were found. The most promi Cancer Cells nent receptor mRNA found in T47D cells was SSTR2, representing about 85% of the whole SSTR mRNA. Characterization of Binding Sites. Fig. 1 presents the binding isotherm of [â€˜251]Tyr'â€˜-somatostatinto whole T47D cells. A biphasic Effect of p-acting Opioids and Somatostatin on Cell Growth saturation curve was obtained with a high-affinity component reach ing saturation (Fig. 1, inset A) and a low-affinity component that did not reach saturation. Nonspecific binding was 27 Â±5% of total Effect of il-acting opioids. Fig. 4 shows the inhibition of cell binding. Analysis of the data in Scatchard coordinates (Fig. 1, inset B) proliferation of T47D human breast cancer cells by morphine and the showed that the binding isotherm could be best fitted in a two-site morphinomimetic peptide morphiceptine. Both p.-acting drugs inhibit model, with affinities of 0.145 Â±0.017 and 1.192 Â±0.093 nr@i,and cell proliferation in a dose-dependent manner, with apparent IC50s of binding capacities of 1450 Â±235 and 11920 Â±1147 sites/cell. This 1.08 and 1.25 nM, respectively. This effect was not inhibited by the heterogeneity of binding of [â€˜251]Tyr'â€˜-somatostatincould indicate opioid antagonist diprenorphine.

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@ ..,...... U) 2t@ .sâ€•@ w @ C, I 0000 0 @. I â€¢ I I 0 2500 5O1X@mOO 10000 0 ru Fig. 1. Binding of [â€˜25IlTyr'â€˜-somatostatinto T47D cells. Saturation binding was performed for 2 Cs) 2.0 h at room temperature on whole cells, as indicated in â€˜Materialsand Methods.â€•The saturation binding 0 i.e isotherm is presented. A, magnification of the mi E @0 tial part of the curve; B, representation of data in 0. u_1.0 Scatchard coordinates. 5000 @T'@@::@a S 0.5 0 0 @n i@m 16000 Bound (pmclesllOÂ° c&1s)

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0 0 20 @[1@ I 2 3 4 5 SSTRtype

Fig. 3. Expression of SSTR mRNA in T47D human breast cancer cells by RT-PCR. Results of RT-PCR of the five 55Th mRNA in T47D cells. Sec â€œMaterialsandMethodsâ€• 0 10.11 10b0 iÃ¸-@ 10.8 j@-l 10.Â° for details. Results are expressed as a percentage of the whole SSTR mRNA detected. Displacer Fig.2. Specificityof [â€˜25I]Tyr'-somatostatinbindingon T47Dcells. Displace ment of [â€˜25lJTyr'â€˜-somatostatinbydifferent analogues. A, 12 fmol of [â€˜25I]Tyr'â€˜- somatostatin (about 50,000 cpm) were incubated with the indicated concentrations of L@1 corticotrophin-releasing hormone (A), somatostatin-14 (s), or somatostatin-28 (â€¢); B, incubated with BIM 23034C, a selective SSTR2 agonist. Points, mean of three experiments in triplicate; bars, SEM. See â€œMaterialsandMethodsâ€•fordetails of the .I-@ binding experiments.

TableI IC50ofdifferentsubstancesonsomatostatinreceptorsofT47Dcells Data were obtained from results presented in Fig. 3. (nM)SomatostatinSubstanceIC@ 1 (nM)IC50 2 140.822.6Somatostatin 280.96.37BIM 23034C5.05 @, 1@ x I- 0.6- Effect of Somatostatin-28 and Sandostatin. Fig. 5 presents the E effects of somatostatin-28 and SMS 201-995 (Sandostatin, a stable z3 0.5 somatostatin analogue) on cell proliferation. Both agents produce a 0.4 dose-dependent inhibition on cell growth. It was of interest that C, maximal suppression of the growth of the cultures was obtained at the 0.3 concentration of 0.1 [email protected] higher concentrations, the inhibition of cell growth was reversed. The concomitant addition of morphine and 0.2 somatostatin did not result in an additive effect, indicating that, probably, both substances might share the same receptor site (not 0.1 shown). 1,11:1.11 Cross-reaction of Morphine with SSTRS 0 -12 -11 -10 -9 -8 -7 -6

To evaluate the possibility that morphine, morphiceptine, and so LogOploid matostatin interact with the same population of receptor sites, we Fig. 4. Effect of @z-actingopioids on cell proliferation of T47D cells. Opioid agonists examined the ability of morphine and morphiceptine to displace (morphine in A and morphiceptine in B) were applied alone in the indicated concentrations [â€˜25I]Tyr'â€˜-somatostatinbinding in T47D cells. Morphiceptine shares (filled columns) or in the presence of l06 M diprenorphine (shaded columns) on T47D cells for 4 days, with daily change of the medium. Cell proliferation was estimated by the the same spectrum as morphine on opioid binding sites in other tetrazolium salt assay, as indicated in â€œMaterialsand Methods.â€•Columns, mean of four systems. Our results are presented in Fig. 6. It can be seen that both experimentsintriplicate;bars,SEM. 5634

further substantiated in the present study by using RT-PCR, showing the expression of SSTR2 (â€”85%)and SSTR3 (â€”15%) in T47D cells. Previous investigations have shown that inhibition of cell prolifer ation by somatostatin analogues might be mediated by SSTR sybtypes SSTR1 and SSTR2 (30). No SSTR1 has been found in the present study. On the contrary, somatostatin-28 and sandostatin, sharing the same affinity for the SSTR2 subtype, decrease the proliferation in T47D cells by the same extent (50 and 55%, respectively), indicating a probable involvement of SSTR2 on the cell growth of T47D cells. The action of somatostatin on cell growth is biphasic. Indeed, maxi mal suppression of growth was obtained at 0. 1 flM.At higher concen trations, the inhibition of cell growth is reversed. The same result has been reported previously on the MCF7 breast cancer cell line by -12 -11 -10 somatostatin (12), antiestrogens (31), and epidermal growth factor (32). Although the underlying mechanism of this biphasic effects is Log Peptide not known, different hypotheses have been proposed, implicating Fig. 5. Effect of somatostatin analogues on cell proliferation of T47D cells. Soma either a desensitization/intemalization/conformational change of the tostatin-28 (SS28, shaded columns) and SMS 201-995 (Sandostatin, filled columns) were receptor or a possible agonist/antagonist effect of the agents. applied in the indicated concentrations on T47D cells for 4 days, with daily change of the medium. Cell proliferation was estimated by the tetrazolium salt assay, as indicated in In a previous study, we have shown that T47D cancer cells express @ â€œMaterialsand Methods.â€•Columns, mean of three experiments in triplicate; bars, SEM. and K opioid receptors. Different opioid agonists (ED-Ala2,D-Leu5]- enkephalin, [D-Ser@,Leu5]-enkephalin, Thr6 etorphine, and ethylke tocyclazocine) selective toward different subtypes of the opioid re morphine and morphiceptine displace the specific binding of ceptor produce a dose-dependent, reversible, inhibitory effect on cell [1 1-somatostatin in a dose-dependent and monophasic man proliferation. Morphine, the p-selective agonist, presenting no affinity ner, similar to BIM 23034C (compare Figs. 2B and 6), with IC50s of for the other opioid receptor subtypes in the T47D cells, produces a 1.67 and 1.71 flM, respectively. major inhibition of cell proliferation, whereas no @topioid receptors have been detected (22). In the present work, we show that this effect DISCUSSION is shared by the amidated tetrapeptide morphiceptine (derived from the enzymatic degradation of j3 casein), which shares, in other sys Various breast cancer cell lines have been found to express SSTRs tems, the same pharmacological selectivity as morphine. In view of (1 1, 12, 20, 21). Addition of somatostatin or somatostatin analogues in the structural homology between the opioid and the SSTRs, which culture media produces a dose-dependent inhibition of cell growth were proposed to derive from the same ancestor gene (5, 6), we have (1 1, 12, 21). In addition, 30% of breast cancers express somatostatin investigated a possible interaction of morphine with the somatostatin immunoreactivity; preneoplastic lesions express less, whereas normal tissue does not show any somatostatin immunoreactivity (14). From the five cloned subtypes of the SSTR, only SSTR2 was found in 5 primary breast tumors, whereas both SSTR2 and SSTR3 were ex pressed in 1 case (16). Furthermore, three breast cancer cell lines with distinct steroid and growth factor receptor patterns (i.e., MCF7, MDA-MB23 1, and ZR-75- 1) express only the SSTR2 subtype, which seems to be the most widely expressed SSTR in different human and rodent tumors or tumor cell lines (27). Using a chemical cross-linking assay, four distinct sites have been identified in the MCF7 cell line in monolayer, one of which binds the somatostatin analogue BIM 23014C (20). In the same study, BIM-23014C identifies one site in MCF7 and T47D cells, three in MDA-MB23 1, and none in the HBL 100 cell line. Finally, the BIM-23014C somatostatin analogue iden tifies at least 1 site in 90% of 30 human breast biopsies. The percent age of somatostatin-positive tumors varies between 14 and 48% (13, C, 28â€”30).Thispercentagebecomes67% when only small tumorsare a C,) selected (29), and 90% with the use of the chemically cross-linked somatostatin analogue and electrophoresis (20). The existence of SSTRs has been reported to be of prognostic value for the evolution of breast cancer (19). The results of the present study indicate the presence of multiple SSTR subtypes in T47D human breast cancer cells. Scatchard analysis of [â€˜25IIJTyr'â€˜-somatostatin-14binding identified two components: one with high affinity but low capacity (Kd, 0. 145 revs;1450 sites/cell), and another oflower affinity but higher capacity (Kd, 1.192 nr@i;11920 0 @t0.h1 lcTbO 10-s 10@8 sites/cell). These results are comparable with those reported by Log Oploid Seytono-Ham et a!. (12) and Prevost et al. (20) on MCF7 cells for the Fig. 6. Displacement of [1251]Tyrt â€˜-somatostatin from its binding sites by jx-acting high-affinity site, and by Weckbecker et al. (21) on the ZR-75-1 opioids. Displacement of [â€˜@I]Tyr'â€˜-somatostatinbinding by the indicated concentrations breast cancer cell lines for the lower affinity site. These results were of morphine (B) or morphiceptine (â€¢).Seeâ€œMaterialsandMethodsâ€•fordetails. 5635

Downloaded from cancerres.aacrjournals.org on September 23, 2021. © 1995 American Association for Cancer Research. MORPHINE AND SSTRs IN T47D BREAST CANCER CELLS ergic system. It was surprising that cross-displacement experiments of effects of somatostatin (analogues) on the growth of human breast cancer cells. somatostatin by morphine and morphiceptine indicated that these Cancer Res., 47: 1566-1570, 1987. 13. Reubi, J. C., Maurer, R., von Werder, J., Torhorst, J., Klijn, J. G. M., and Lamberts, opioids compete for somatostatin binding in a similar manner, inhib S. W. J. Somatostatinreceptorsin humanendocrinetumors.Cancer Res., 47: iting cell proliferation through an interaction with SSTR2. The expo 551â€”558,1987. 14. Ciocca, D. R., Puy, L. A., Fasoli, L. C., Tello, 0., Aznar, J. C., Gago, F. E., Papa, S. I., sure of cells to both somatostatin and morphine does not produce and Sonego, R. Corticotropin releasing hormone, luteinizing hormone-releasing hor additive effects, enhancing the assumption that these substances might mone, growth hormone-releasing hormone, and somatostatin-like immunoreactivities interact with the same receptor site. From these data, we have con in biopsies from breast cancer patients. Breast Cancer Res. Treat.. 15: 175-184, 1990. 15. Scopsi, L., Balslev, E., Brunner, N., Skovgaart-Poulsen, H., Andersen, J., Rank, F., cluded that morphine may exert its antiproliferative action through and Larson, L-I. Immunoreactive opioid peptides in human breast cancer. Am. J. SSTR2. Pathol.,134:473â€”479,1989. Previous studies have indicated a possible interaction between 16. Reubi, J. C., Schaer, J. C., Waser, B., and Mengod, G. Expression and localization of somatostatin receptor SSTRI, SSTR2, and SSTR3 messenger RNA in primary human somatostatin analogues and the opioid receptor system. Indeed, tumors using in situ hybridization. Cancer Res., 54: 3455â€”3459, 1994. Maurer et a!. (33) and Walker et a!. (34) have reported that the 17. Reubi, J. C., Krenning, E., Lamberts, S. W. J., and Kvols, L. In vitro detection of octapeptide analogues of somatostatin act, with high affinities, as somatostatin receptors in human tumors. Metabolism, 41 (Suppl. 2): 104â€”110,1992. 18. Srkalovic, G., Cai, R. Z., and Schally, A. V. Evaluation of receptors for somatostatin antagonists to @.topioid receptors in the central nervous system. in various tumors using different analogs. J. Clin. Endocrinol. Metab., 70: 661â€”669, Although the high degree of homology between the opioid and SSTRs 1990. 19. Foekens,J.A., Portengen,H.,vanPutten,W.L. J., Trapman.A.M. A. C., Reubi, suggests that some ligands might be able to interact with both recep J-C., Alexieva-Figusch, J., and Klijn, J. G. M. Prognostic value of receptors for tors (6), to our knowledge, no cross-reaction of opiates with SSTRs insulin-like growth factor I, somatostatin, and epidermal growth factor in human has been described previously. Furthermore, from our studies and breast cancer. Cancer Res., 49: 7002â€”7009, 1989. 20. Prevost, G., Lanson, M., Thomas, F., Veber, N., Gonzalez, W., Beaupain, R., Stance, those of others (30), it appears that SSTR2 may be responsible for the A.,andBogden,A.MolecularheterogeneityofsomatostatinanalogueBIM-230'4C mediation of the observed antiproliferative effect. Together, the data receptors in human breast carcinoma cells using the chemical cross-linking assay. presented here provide evidence for a functional interaction of the two Cancer Res., 52: 843â€”850, 1992. 21. Weckbecker, G. Liu, R., Tolcsvai, L., and Bruns, C. Antiproliferative effects of the major inhibitory neuroendocrine systems (opioids and somatostatin), somatostatin analogue octreotide (SMS 201â€”995)on ZR-75-l human breast cancer which may involve peptide competition and may have implications cells in vivo and in vitro. Cancer Res., 52: 4973â€”4978, 1992. for the biology and pathophysiology of tumor cells and disease states. 22. Hatzoglou, A., Bakogeorgou, E., and Castanas, E. The antiproliferative effect of opioid agonists on the T47D human breast cancer cell line, is partially mediated In addition, the fact that morphiceptine, a casomorphin produced from through opioid receptors. Eur. J. Pharmacol., in press, 1995. 13casein(a normalproductof breastepithelium),bindsto SSTRsand 23. Maneckjee, R., Biswas, R., and Vonderhaar, B. K. Binding of opioids to human MCF-7 breast cancer cells and their effects on growth. 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Downloaded from cancerres.aacrjournals.org on September 23, 2021. © 1995 American Association for Cancer Research. Morphine Cross-Reacts with Somatostatin Receptor SSTR2 in the T47D Human Breast Cancer Cell Line and Decreases Cell Growth

Anastassia Hatzoglou, L'Houcine Ouafik, Efstathia Bakogeorgou, et al.

Cancer Res 1995;55:5632-5636.

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