Abstract
The major challenge in treating cancers with ATRA is the limited availability inside the cell and resistance developed in prolonged treatment. We made an attempt for co-treatment of human NSCLC cell lines (A549) with ATRA and its isomeric precursor (9cisRA). In this study, the growth inhibitory effect of ATRA, 9cisRA and combination of both were tested in A549 cells by MTT and Trypan blue assays. As the effects of retinoid are mediated through their receptors, their gene expression levels were analyzed by RT-PCR. The target gene receptor, RAR-β protein expression, was analyzed by immunocytochemistry. The cancer cell (A549) growth inhibitory effect was significantly (p ≤ 0.001) enhanced in combination treatment when compared with the result of individual treatments. The mRNA expression levels of both RAR-β and RXR-β were found to be increased in co-treatment (band density of 0.75 and 0.806, respectively) when compared with 9cisRA treatment (0.25 and 0.112) and ATRA treatment (0.01 and 0.081). A concomitant enhancement in the target RAR-β protein expression was observed in co-treated cells when compared with individual treatments. We thus conclude that the co-treatment had increased the availability of ATRA, by isomerization of the 9cisRA which then resulted in an increased expression of both RAR-β and RXR-β receptors and the target protein RAR-β which in turn inhibited lung cancer cell growth. Our study results have explored the mechanism of synergistic effect of co-treatment with ATRA and 9cisRA and further preclinical studies are necessary to validate the application of co-treatment of retinoid in clinical use.
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References
Altucci L, Leibowitz MD, Ogilvie KM, De Lera AR, Gronemeyer H (2007) RAR and RXR modulation in cancer and metabolic disease. Nat Rev Drug Discovery 6(10):793–810
Bogos K, Renyi-Vamos F, Kovacs G, Tovari J, Dome B (2008) Role of retinoic receptors in lung carcinogenesis. J Exp Clin Cancer Res 27(1):18
Brabender J, Metzger R, Salonga D, Danenberg KD, Danenberg PV, Hölscher AH, Schneider PM (2005) Comprehensive expression analysis of retinoic acid receptors and retinoid X receptors in non-small cell lung cancer: implications for tumor development and prognosis. Carcinogenesis 26(3):525–530
Bushue N, Wan Y-JY (2010) Retinoid pathway and cancer therapeutics. Adv Drug Deliv Rev 62(13):1285–1298
Buskohl PR, Gould RA, Curran S, Archer SD, Butcher JT (2012) Multidisciplinary inquiry-based investigation learning using an ex vivo chicken culture platform: role of vitamin A on embryonic morphogenesis. Am Biol Teacher 74(9):636–643
Cheung B, Yan J, Smith SA, Nguyen T, Lee M, Kavallaris M, Norris MD, Haber M, Marshall GM (2003) Growth inhibitory retinoid effects after recruitment of retinoid X receptor β to the retinoic acid receptor β promoter. Int J Cancer 105(6):856–867
Cras A, Darsin-Bettinger D, Balitrand N, Cassinat B, Soulie A, Toubert M, Delva L, Chomienne C (2007) Epigenetic patterns of the retinoic acid receptor β2 promoter in retinoic acid-resistant thyroid cancer cells. Oncogene 26(27):4018–4024
Di Masi A, Leboffe L, De Marinis E, Pagano F, Cicconi L, Rochette-Egly C, Lo-Coco F, Ascenzi P, Nervi C (2015) Retinoic acid receptors: from molecular mechanisms to cancer therapy. Mol Aspects Med 41:1–115
Dragnev KH, Rigas JR, Dmitrovsky E (2000) The retinoids and cancer prevention mechanisms. Oncologist 5(5):361–368
Han S, Fukazawa T, Yamatsuji T, Matsuoka J, Miyachi H, Maeda Y, Durbin M, Naomoto Y (2010) Anti-tumor effect in human lung cancer by a combination treatment of novel histone deacetylase inhibitors: SL142 or SL325 and retinoic acids. PLoS ONE 5(11):e13834
Hanahan D, Weinberg RA (2000) The hallmarks of cancer. Cell 100(1):57–70
Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D (2011) Global cancer statistics. Cancer J Clin 61(2):69–90
Lei M, de Thé H (2003) Retinoids and retinoic acid receptor in cancer. Eur J Cancer Suppl 1(2):13–18
Lotan R (1999) Aberrant expression of retinoid receptors and lung carcinogenesis. Oxford University Press, Oxford
Mi L, Wang X, Govind S, Hood BL, Veenstra TD, Conrads TP, Saha DT, Goldman R, Chung F-L (2007) The role of protein binding in induction of apoptosis by phenethyl isothiocyanate and sulforaphane in human non-small lung cancer cells. Can Res 67(13):6409–6416
Minna JD, Roth JA, Gazdar AF (2002) Focus on lung cancer. Cancer Cell 1(1):49–52
Moison C, Senamaud-Beaufort C, Fourrière L, Champion C, Ceccaldi A, Lacomme S, Daunay A, Tost J, Arimondo PB, Guieysse-Peugeot A-L (2013) DNA methylation associated with polycomb repression in retinoic acid receptor β silencing. FASEB J 27(4):1468–1478
Pavan B, Dalpiaz A, Biondi C, Nieddu M, De Luca A, Prasad PD, Paganetto G, Favaloro B (2008) An RPE cell line as a useful in vitro model for studying retinoic acid receptor β: expression and affinity. Biosci Rep 28(6):327–334
Pettersson F, Dalgleish A, Bissonnette R, Colston K (2002) Retinoids cause apoptosis in pancreatic cancer cells via activation of RAR-γ and altered expression of Bcl-2/Bax. Br J Cancer 87(5):555–561
Ramya D, Siddikuzzaman Manjamalai A, Berlin Grace V (2012) Chemoprotective effect of all-trans retinoic acid (ATRA) on oxidative stress and lung metastasis induced by benzo (a) pyrene. Immunopharmacol Immunotoxicol 34(2):317–325
Rhinn M, Dollé P (2012) Retinoic acid signalling during development. Development 139(5):843–858
Siddikuzzaman, Grace VB (2012) Inhibition of metastatic lung cancer in C57BL/6 mice by liposome encapsulated all trans retinoic acid (ATRA). Int Immunopharmacol 14(4):570–579
Soria J-C, Xu X, Liu DD, Lee JJ, Kurie J, Morice RC, Khuri F, Mao L, Hong WK, Lotan R (2003) Retinoic acid receptor β and telomerase catalytic subunit expression in bronchial epithelium of heavy smokers. J Natl Cancer Inst 95(2):165–168
Stewart B, Wild CP (2017) World cancer report 2014. Health
Sun S-Y, Wan H, Yue P, Hong WK, Lotan R (2000) Evidence that retinoic acid receptor β induction by retinoids is important for tumor cell growth inhibition. J Biol Chem 275(22):17149–17153
Szabó DR, Baghy K, Szabó PM, Zsippai A, Marczell I, Nagy Z, Varga V, Éder K, Tóth S, Buzás EI (2014) Antitumoral effects of 9-cis retinoic acid in adrenocortical cancer. Cell Mol Life Sci 71(5):917–932
Wen X, Li Y, Hu K, Dai C, Liu Y (2005) Hepatocyte growth factor receptor signaling mediates the anti-fibrotic action of 9-cis-retinoic acid in glomerular mesangial cells. Am J Pathol 167(4):947–957
Zuccari G, Carosio R, Fini A, Montaldo P, Orienti I (2005) Modified polyvinylalcohol for encapsulation of all-trans-retinoic acid in polymeric micelles. J Control Release 103(2):369–380
Acknowledgements
The authors would like to acknowledge the Karunya Institute of Technology and Sciences for the financial support given through Karunya Short-Term Project Grant. We also acknowledge partial financial support shared from DST-SERB and DBT, Govt. of India. We would also like to acknowledge the valuable technical support of Dr. C. Vani, Assistant Professor and Ms. Jissin Mathew, Research Scholar, Department of Biotechnology, Karunya Institute of Technology and Sciences, Coimbatore.
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Bama, E.S., Grace, V.M.B., Sundaram, V. et al. Synergistic effect of co-treatment with all-trans retinoic acid and 9-cis retinoic acid on human lung cancer cell line at molecular level. 3 Biotech 9, 159 (2019). https://doi.org/10.1007/s13205-019-1692-x
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DOI: https://doi.org/10.1007/s13205-019-1692-x