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19-09-2023 | Salbutamol | Original Article

The Beta2-adrenergic agonist salbutamol synergizes with paclitaxel on cell proliferation and tumor growth in triple negative breast cancer models

Authors: Martina Jabloñski, María Sol Rodríguez, Ezequiel Mariano Rivero, Carlos David Bruque, Silvia Vanzulli, Ariana Bruzzone, Cecilia Pérez Piñero, Isabel Alicia Lüthy

Published in: Cancer Chemotherapy and Pharmacology | Issue 6/2023

Abstract

Purpose

Globally breast cancer accounts for 24.5% in incidence and 15.5% in cancer deaths in women. The triple-negative subtype lacks any specific therapy and is treated with chemotherapy, resulting in significant side-effects. We aimed to investigate if the dose of chemotherapeutic drugs could be diminished by co-administering it with the β2-agonist salbutamol.

Methods

Cell proliferation was measured by thymidine incorporation; gene expression, by real-time PCR and protein phosphorylation by WB. Apoptosis was assessed by acridine orange / ethidium bromide and TUNEL tests. Public patient databases were consulted. Cells were inoculated to nude mice and their growth assessed.

Results

The β₂-agonist salbutamol synergizes in MDA-MB-231 cells in vitro with paclitaxel and doxorubicin on cell proliferation through ADRB2 receptors, while the β-blocker propranolol does not. The expression of this receptor was assessed in patient databases and other cell lines. Triple negative samples had the lowest expression. Salbutamol and paclitaxel decreased MDA-MB-231 cell proliferation while their combination further inhibited it. The pathways involved were analyzed. When these cells were inoculated to nude mice, paclitaxel and salbutamol inhibited tumor growth. The combined effect was significantly greater. Paclitaxel increased the expression of MDR1 while salbutamol partially reversed this increase.

Conclusion

While the effect of salbutamol was mainly on cell proliferation, suboptimal concentrations of paclitaxel provoked a very important enhancement of apoptosis. The latter enhanced transporter proteins as MDR1, whose expression were diminished by salbutamol. The expression of ADRB2 should be assessed in the biopsy or tumor to eventually select patients that could benefit from salbutamol repurposing.

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Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, Bray F (2021) Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 71:209–249. https://doi.org/10.3322/caac.21660CrossRefPubMed

Perou CM, Sorlie T, Eisen MB, van de Rijn M, Jeffrey SS, Rees CA, Pollack JR, Ross DT, Johnsen H, Akslen LA, Fluge O, Pergamenschikov A, Williams C, Zhu SX, Lonning PE, Borresen-Dale AL, Brown PO, Botstein D (2000) Molecular portraits of human breast tumours. Nature 406:747–752. https://doi.org/10.1038/35021093CrossRefPubMed

Prat A, Perou CM (2011) Deconstructing the molecular portraits of breast cancer. Mol Oncol 5:5–23. https://doi.org/10.1016/j.molonc.2010.11.003CrossRefPubMed

Sharifi-Rad J, Quispe C, Patra JK, Singh YD, Panda MK, Das G, Adetunji CO, Michael OS, Sytar O, Polito L, Zivkovic J, Cruz-Martins N, Klimek-Szczykutowicz M, Ekiert H, Choudhary MI, Ayatollahi SA, Tynybekov B, Kobarfard F, Muntean AC, Grozea I, Dastan SD, Butnariu M, Szopa A, Calina D (2021) Paclitaxel: application in modern oncology and nanomedicine-based cancer therapy. Oxid Med Cell Longev 2021:3687700. https://doi.org/10.1155/2021/3687700CrossRefPubMedPubMedCentral

Kamgar M, Greenwald MK, Assad H, Hastert TA, McLaughlin EM, Reding KW, Paskett ED, Bea JW, Shadyab AH, Neuhouser ML, Nassir R, Crane TE, Sreeram K, Simon MS (2021) Prevalence and predictors of peripheral neuropathy after breast cancer treatment. Cancer Med 10:6666–6676. https://doi.org/10.1002/cam4.4202CrossRefPubMedPubMedCentral

Draguet A, Tagliatti V, Colet JM (2021) Targeting metabolic reprogramming to improve breast cancer treatment: an in vitro evaluation of selected metabolic inhibitors using a metabolomic approach. Metabolites. https://doi.org/10.3390/metabo11080556.10.3390/metabo11080556CrossRefPubMedPubMedCentral

Ma X, Hu Y, Batebi H, Heng J, Xu J, Liu X, Niu X, Li H, Hildebrand PW, Jin C, Kobilka BK (2020) Analysis of Beta2ar-Gs and Beta2ar-Gi complex formation by NMR spectroscopy. Proc Natl Acad Sci USA 117:23096–23105. https://doi.org/10.1073/pnas.2009786117CrossRefPubMedPubMedCentral

Vandewalle B, Revillion F, Lefebvre J (1990) Functional beta-adrenergic receptors in breast cancer cells. J Cancer Res Clin Oncol 116:303–306CrossRefPubMed

Marchetti B, Spinola PG, Plante M, Poyet P, Follea N, Pelletier G, Labrie F (1989) Beta-adrenergic receptors in DMBA-induced rat mammary tumors: correlation with progesterone receptor and tumor growth. Breast Cancer Res Treat 13:251–263CrossRefPubMed

10.

Draoui A, Vandewalle B, Hornez L, Revillion F, Lefebvre J (1991) Beta-adrenergic receptors in human breast cancer: identification, characterization and correlation with progesterone and estradiol receptors. Anticancer Res 11:677–680PubMed

11.

Marchetti B, Spinola PG, Pelletier G, Labrie F (1991) A potential role for catecholamines in the development and progression of carcinogen-induced mammary tumors: hormonal control of beta-adrenergic receptors and correlation with tumor growth. J Steroid BiochemMolBiol 38:307–320CrossRef

12.

Chang A, Le CP, Walker AK, Creed SJ, Pon CK, Albold S, Carroll D, Halls ML, Lane JR, Riedel B, Ferrari D, Sloan EK (2016) Beta2-Adrenoceptors on tumor cells play a critical role in stress-enhanced metastasis in a mouse model of breast cancer. Brain Behav Immun 57:106–115. https://doi.org/10.1016/j.bbi.2016.06.011CrossRefPubMedPubMedCentral

13.

Sloan EK, Priceman SJ, Cox BF, Yu S, Pimentel MA, Tangkanangnukul V, Arevalo JM, Morizono K, Karanikolas BD, Wu L, Sood AK, Cole SW (2010) The sympathetic nervous system induces a metastatic switch in primary breast cancer. Cancer Res 70:7042–7052. https://doi.org/10.1158/0008-5472.CAN-10-0522CrossRefPubMedPubMedCentral

14.

Hara MR, Kovacs JJ, Whalen EJ, Rajagopal S, Strachan RT, Grant W, Towers AJ, Williams B, Lam CM, Xiao K, Shenoy SK, Gregory SG, Ahn S, Duckett DR, Lefkowitz RJ (2011) A stress response pathway regulates DNA damage through Beta2-adrenoreceptors and beta-arrestin-1. Nature 477:349–353. https://doi.org/10.1038/nature10368CrossRefPubMedPubMedCentral

15.

Powe DG, Entschladen F (2011) Targeted therapies: using beta-blockers to inhibit breast cancer progression. Nat Rev Clin Oncol 8:511–512CrossRefPubMed

16.

Carie AE, Sebti SM (2007) A chemical biology approach identifies a Beta-2 adrenergic receptor agonist that causes human tumor regression by blocking the Raf-1/Mek-1/Erk1/2 pathway. Oncogene 26:3777–3788CrossRefPubMed

17.

Perez Pinero C, Bruzzone A, Sarappa MG, Castillo LF, Luthy IA (2012) Involvement of Alpha2- and Beta2-adrenoceptors on breast cancer cell proliferation and tumour growth regulation. Br J Pharmacol 166:721–736. https://doi.org/10.1111/j.1476-5381.2011.01791.xCrossRefPubMedPubMedCentral

18.

Rivero EM, Pinero CP, Gargiulo L, Entschladen F, Zanker K, Bruzzone A, Luthy IA (2017) The Beta 2-adrenergic agonist salbutamol inhibits migration, invasion and metastasis of the human breast cancer MDA-MB-231 cell line. Curr Cancer Drug Targets 17:756–766CrossRefPubMed

19.

Slotkin TA, Zhang J, Dancel R, Garcia SJ, Willis C, Seidler FJ (2000) Beta-adrenoceptor signaling and its control of cell replication in MDA-MB-231 human breast cancer cells. Breast Cancer Res Treat 60:153–166CrossRefPubMed

20.

Gargiulo L, Copsel S, Rivero EM, Gales C, Senard JM, Luthy IA, Davio C, Bruzzone A (2014) Differential Beta(2)-adrenergic receptor expression defines the phenotype of non-tumorigenic and malignant human breast cell lines. Oncotarget 5:10058–10069CrossRefPubMedPubMedCentral

21.

Rivero EM, Martinez LM, Bruque CD, Gargiulo L, Bruzzone A, Lüthy IA (2019) Prognostic significance of α2 and β2-adrenoceptor gene expression in breast cancer patients. Br J Clin Pharmacol 85:2143–2154. https://doi.org/10.1111/bcp.14030CrossRefPubMedPubMedCentral

22.

Barron TI, Connolly RM, Sharp L, Bennett K, Visvanathan K (2011) Beta blockers and breast cancer mortality: a population-based study. J Clin Oncol 29:2635–2644. https://doi.org/10.1200/JCO.2010.33.5422CrossRefPubMed

23.

Gargiulo L, Rivero EM, di Siervi N, Buzzi ED, Buffone MG, Davio CA, Luthy IA, Bruzzone A (2020) Agonist effects of propranolol on non-tumor human breast cells. Cells 9:1036. https://doi.org/10.3390/cells9041036CrossRefPubMedPubMedCentral

24.

Vazquez SM, Mladovan AG, Perez C, Bruzzone A, Baldi A, Luthy IA (2006) Human breast cell lines exhibit functional Alpha(2)-adrenoceptors. Cancer Chemother Pharmacol 58:50–61CrossRefPubMed

25.

Ribble D, Goldstein NB, Norris DA, Shellman YG (2005) A simple technique for quantifying apoptosis in 96-well plates. BMC Biotechnol 5:12. https://doi.org/10.1186/1472-6750-5-12CrossRefPubMedPubMedCentral

26.

Bruzzone A, Pinero CP, Castillo LF, Sarappa MG, Rojas P, Lanari C, Luthy IA (2008) Alpha(2)-Adrenoceptor action on cell proliferation and mammary tumour growth in mice. Br J Pharmacol 155:494–504. https://doi.org/10.1038/bjp.2008.278CrossRefPubMedPubMedCentral

27.

Gargiulo L, May M, Rivero EM, Copsel S, Lamb C, Lydon J, Davio C, Lanari C, Luthy IA, Bruzzone A (2017) A novel effect of beta-adrenergic receptor on mammary branching morphogenesis and its possible implications in breast cancer. J Mammary Gland Biol Neoplasia 22:43–57. https://doi.org/10.1007/s10911-017-9371-1CrossRefPubMed

28.

Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254. https://doi.org/10.1006/abio.1976.9999CrossRefPubMed

29.

Institute of Laboratory Animal Resources Commission on Life Sciences, National Research Council (1996) Guide for the care and use of laboratory animals. National Academy Press, Washington, DC

30.

Institute of Laboratory Animal Resources Commission on Life SNR, Council (2010) Guide for the care and use of laboratory animals: eighth edition. National Academy Press, Washington, DC

31.

United Kingdom Co-ordinating Committee on Cancer Research U (1998) United Kingdom Co-ordinating Committee on Cancer Research (UKCCCR) guidelines for the welfare of animals in experimental neoplasia (Second Edition). Br J Cancer 77:1–10

32.

Workman P, Aboagye EO, Balkwill F, Balmain A, Bruder G, Chaplin DJ, Double JA, Everitt J, Farningham DA, Glennie MJ, Kelland LR, Robinson V, Stratford IJ, Tozer GM, Watson S, Wedge SR, Eccles SA (2010) Guidelines for the welfare and use of animals in cancer research. Br J Cancer 102:1555–1577. https://doi.org/10.1038/sj.bjc.66056426605642CrossRefPubMedPubMedCentral

33.

Chen L, Ye HL, Zhang G, Yao WM, Chen XZ, Zhang FC, Liang G (2014) Autophagy inhibition contributes to the synergistic interaction between EGCG and doxorubicin to kill the hepatoma Hep3B cells. PLoS ONE 9:e85771. https://doi.org/10.1371/journal.pone.0085771CrossRefPubMedPubMedCentral

34.

Wackerhage H, Christensen JF, Ilmer M, von Luettichau I, Renz BW, Schonfelder M (2022) Cancer catecholamine conundrum. Trends Cancer 8:110–122. https://doi.org/10.1016/j.trecan.2021.10.005CrossRefPubMed

35.

Marques L, Vale N (2022) Salbutamol in the management of asthma: a review. Int J Mol Sci. https://doi.org/10.3390/ijms232214207CrossRefPubMedPubMedCentral

36.

Emeryk A, Emeryk-Maksymiuk J (2020) Short-acting inhaled Beta2-agonists: Why, whom, what, how? Adv Respir Med 88:443–449. https://doi.org/10.5603/ARM.a2020.0132CrossRefPubMed

37.

Casabella-Ramon S, Jimenez-Sabado V, Tarifa C, Casellas S, Lu TT, Izquierdo-Castro P, Gich I, Jimenez M, Ginel A, Guerra JM, Chen SRW, Benitez R, Hove-Madsen L (2022) Impact of R-carvedilol on Beta2-adrenergic receptor-mediated spontaneous calcium release in human atrial myocytes. Biomedicines. https://doi.org/10.3390/biomedicines10071759CrossRefPubMedPubMedCentral

38.

Gebauer L, Arul Murugan N, Jensen O, Brockmoller J, Rafehi M (2022) Molecular basis for stereoselective transport of fenoterol by the organic cation transporters 1 and 2. Biochem Pharmacol 197:114871. https://doi.org/10.1016/j.bcp.2021.114871CrossRefPubMed

39.

van der Westhuizen ET, Breton B, Christopoulos A, Bouvier M (2014) Quantification of ligand bias for clinically relevant Beta2-adrenergic receptor ligands: implications for drug taxonomy. Mol Pharmacol 85:492–509CrossRefPubMed

40.

Pasquier E, Ciccolini J, Carre M, Giacometti S, Fanciullino R, Pouchy C, Montero MP, Serdjebi C, Kavallaris M, Andre N (2011) Propranolol potentiates the anti-angiogenic effects and anti-tumor efficacy of chemotherapy agents: implication in breast cancer treatment. Oncotarget 2:797–809CrossRefPubMedPubMedCentral

41.

Botteri E, Munzone E, Rotmensz N, Cipolla C, De Giorgi V, Santillo B, Zanelotti A, Adamoli L, Colleoni M, Viale G, Goldhirsch A, Gandini S (2013) Therapeutic effect of beta-blockers in triple-negative breast cancer postmenopausal women. Breast Cancer Res Treat 140:567–575. https://doi.org/10.1007/s10549-013-2654-3CrossRefPubMed

42.

Melhem-Bertrandt A, Chavez-Macgregor M, Lei X, Brown EN, Lee RT, Meric-Bernstam F, Sood AK, Conzen SD, Hortobagyi GN, Gonzalez-Angulo AM (2011) Beta-blocker use is associated with improved relapse-free survival in patients with triple-negative breast cancer. J Clin Oncol 29:2645–2652. https://doi.org/10.1200/JCO.2010.33.4441CrossRefPubMedPubMedCentral

43.

Lofling LL, Stoer NC, Sloan EK, Chang A, Gandini S, Ursin G, Botteri E (2022) Beta-blockers and breast cancer survival by molecular subtypes: a population-based cohort study and meta-analysis. Br J Cancer 127:1086–1096. https://doi.org/10.1038/s41416-022-01891-7CrossRefPubMedPubMedCentral

44.

Chang A, Botteri E, Gillis RD, Lofling L, Le CP, Ziegler AI, Chung NC, Rowe MC, Fabb SA, Hartley BJ, Nowell CJ, Kurozumi S, Gandini S, Munzone E, Montagna E, Eikelis N, Phillips SE, Honda C, Masuda K, Katayama A, Oyama T, Cole SW, Lambert GW, Walker AK, Sloan EK (2023) Beta-blockade enhances anthracycline control of metastasis in triple-negative breast cancer. Sci Transl Med 15:eadf1147. https://doi.org/10.1126/scitranslmed.adf1147CrossRefPubMed

45.

Serini S, Calviello G (2017) Modulation of Ras/ERK and phosphoinositide signaling by long-chain n-3 PUFA in breast cancer and their potential complementary role in combination with targeted drugs. Nutrients. https://doi.org/10.3390/nu9030185CrossRefPubMedPubMedCentral

46.

Bruzzone A, Sauliere A, Finana F, Senard JM, Luthy I, Gales C (2014) Dosage-dependent regulation of cell proliferation and adhesion through dual Beta2-adrenergic receptor/cAMP signals. FASEB J 28:1342–1354. https://doi.org/10.1096/fj.13-239285CrossRefPubMed

47.

Amin T, Sharma RP, Mir KB, Slathia N, Chhabra S, Tsering D, Kotwal P, Bhagat M, Nandi U, Parkesh R, Kapoor KK, Goswami A (2023) Quinoxalinone substituted pyrrolizine (4h)-induced dual inhibition of AKT and ERK instigates apoptosis in breast and colorectal cancer by modulating mitochondrial membrane potential. Eur J Pharmacol. https://doi.org/10.1016/j.ejphar.2023.175945CrossRefPubMedPubMedCentral

48.

Montoya A, Amaya CN, Belmont A, Diab N, Trevino R, Villanueva G, Rains S, Sanchez LA, Badri N, Otoukesh S, Khammanivong A, Liss D, Baca ST, Aguilera RJ, Dickerson EB, Torabi A, Dwivedi AK, Abbas A, Chambers K, Bryan BA, Nahleh Z (2017) Use of non-selective beta-blockers is associated with decreased tumor proliferative indices in early stage breast cancer. Oncotarget 8:6446–6460. https://doi.org/10.18632/oncotarget.14119CrossRefPubMed

49.

Garona J, Pifano M, Pastrian MB, Gomez DE, Ripoll GV, Alonso DF (2016) Addition of vasopressin synthetic analogue [V(4)Q(5)]dDAVP to standard chemotherapy enhances tumour growth inhibition and impairs metastatic spread in aggressive breast tumour models. Clin Exp Meta 33:589–600. https://doi.org/10.1007/s10585-016-9799-5CrossRef

50.

Stribbling SM, Ryan AJ (2022) The cell-line-derived subcutaneous tumor model in preclinical cancer research. Nat Protoc 17:2108–2128. https://doi.org/10.1038/s41596-022-00709-3CrossRefPubMed

51.

Killion JJ, Radinsky R, Fidler IJ (1998) Orthotopic models are necessary to predict therapy of transplantable tumors in mice. Cancer Meta Rev 17:279–284. https://doi.org/10.1023/a:1006140513233CrossRef

52.

Mayati A, Moreau A, Denizot C, Stieger B, Parmentier Y, Fardel O (2017) Beta2-adrenergic receptor-mediated in vitro regulation of human hepatic drug transporter expression by epinephrine. Eur J Pharmaceut Sci Off J Eur Feder Pharmaceut Sci 106:302–312. https://doi.org/10.1016/j.ejps.2017.06.010CrossRef

53.

Yao H, Duan Z, Wang M, Awonuga AO, Rappolee D, Xie Y (2009) Adrenaline induces chemoresistance in HT-29 colon adenocarcinoma cells. Cancer Genet Cytogenet 190:81–87. https://doi.org/10.1016/j.cancergencyto.2008.12.009CrossRefPubMed

54.

Liang Y, Wu G, Luo T, Xie H, Zuo Q, Huang P, Li H, Chen L, Lu H, Chen Q (2023) 10-Gingerol enhances the effect of taxol in triple-negative breast cancer via targeting ADRB2 signaling. Drug Des Devel Ther 17:129–142. https://doi.org/10.2147/DDDT.S390602CrossRefPubMedPubMedCentral

55.

Abu Samaan TM, Samec M, Liskova A, Kubatka P, Busselberg D (2019) Paclitaxel’s mechanistic and clinical effects on breast cancer. Biomolecules 9:789. https://doi.org/10.3390/biom9120789CrossRefPubMedPubMedCentral

Title: The Beta2-adrenergic agonist salbutamol synergizes with paclitaxel on cell proliferation and tumor growth in triple negative breast cancer models
Authors: Martina Jabloñski
María Sol Rodríguez
Ezequiel Mariano Rivero
Carlos David Bruque
Silvia Vanzulli
Ariana Bruzzone
Cecilia Pérez Piñero
Isabel Alicia Lüthy
Publication date: 19-09-2023
Publisher: Springer Berlin Heidelberg
Keywords: Salbutamol
Breast Cancer
Breast Cancer
Propranolol
Published in: Cancer Chemotherapy and Pharmacology / Issue 6/2023
Print ISSN: 0344-5704
Electronic ISSN: 1432-0843
DOI: https://doi.org/10.1007/s00280-023-04586-9

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