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Current Topics in Medicinal Chemistry


ISSN (Print): 1568-0266
ISSN (Online): 1873-4294

Research Article

Cedrus deodara (Bark) Essential Oil Induces Apoptosis in Human Colon Cancer Cells by Inhibiting Nuclear Factor kappa B

Author(s): Madhulika Bhagat*, Ajay Kumar and Renuka Suravajhala

Volume 20 , Issue 22 , 2020

Page: [1981 - 1992] Pages: 12

DOI: 10.2174/1568026620666200722120826

Price: $65


Aims: The aim of this study is to explore essential oil from the bark of Cedrus deodara (CDEO) as an potential anticancer agent.

Background: The frontline drugs against cancer in clinical settings are posing challenges of resistance and other detrimental side-effects. This has led to the exploration of new anticancer chemical entities from natural sources, particularly plant-based products such as essential oils that serve as vast repositories of pharmacologically active substances for combating cancer.

Objective: The objective is to isolate and characterize the essential oil from the bark of Cedrus deodara (CDEO) and evaluate its potential as an anticancer agent and delineate the possible underlying mechanism of action.

Methods: Cedrus deodara essential oil from bark (CDEO) was obtained by hydro-distillation and analyzed by GC/MS for vital constituents. Further, in vitro cytotoxic potential was measured by MTT assay against a panel of cancer cell lines. The apoptosis-inducing potential of CDEO was analyzed by mitochondrial membrane potential loss (ΔΨm) and nuclear fragmentation assay. Besides, wound healing assay and colonogenic assay were employed to check the anti-metastatic potential of CDEO. Molecular docking approaches were employed for target identification, while immuno-blotting was carried out for target validation.

Results and Discussion: The major components identified were 2-(tert-Buyl)-6-methyl-3-(2- (trifluoromethyl) benzyl)imidazo [1,2-a]pyridine (26.32 %);9- Octadecenoic acid (8.015 %); Copaene (5.181 %);2-(4-Methoxy-2,6-dimethylphenyl) -3-methyl-2H- benzo[g]indazole(4.36 %) and 9(E),11(E)- Conjugated linoleic acid (4.299 %). Further, potent in vitro cytotoxic activity with IC50 values of 11.88 μg/ ml and 14.63 μg/ ml in colon cancer cell lines of HCT-116 and SW-620, respectively. Further, a significant and dose-dependent decrease in colony formation, cell migration, induction of ROS formation and loss in ΔΨm was observed. Additionally, major compounds identified were chosen for ligandprotein binding interaction studies to predict the molecular targets in colon cancer. It was observed that compounds such as 9-Octadecenoic acid;4H-1- Benzopyran-4-one, 3-(3,4-dimethoxyphenyl)-6,7- dimethoxy; 2-(4-Methoxy-2,6-dimethylphenyl) -3-methyl-2H-benzo [g]indazole and 2-Bornanol,5-(2,4- dinitro phenyl) hydrazono have a prominent binding affinity with NF-κB. This was also further validated by immuno-blotting results wherein CDEO treatment in colon cancer cells led to the abrogation of NFκB, and the Bcl-2-associated X protein (Bax): B-cell lymphoma (Bcl)-2 ratio was up-regulated leading to enhanced cleaved caspase 3 formation and subsequent apoptosis.

Conclusion: These results unveil CDEO inhibits cell proliferation and induces apoptosis in colon cancer cells, which can be attributed to the abrogation of the NFκB signaling pathway.

Keywords: Cedrus deodara, Essential oil, Colon cancer, Antiproliferative, Apoptosis, Cell migration.

Graphical Abstract
Sitarz, R.; Skierucha, M.; Mielko, J.; Offerhaus, G.J.A.; Maciejewski, R.; Polkowski, W.P. Gastric cancer: epidemiology, prevention, classification, and treatment. Cancer Manag. Res., 2018, 10, 239-248.
[] [PMID: 29445300]
Bhandari, A.; Woodhouse, M.; Gupta, S. Colorectal cancer is a leading cause of cancer incidence and mortality among adults younger than 50 years in the USA: a SEER-based analysis with comparison to other young-onset cancers. J. Investig. Med., 2017, 65(2), 311-315.
[] [PMID: 27864324]
Aqil, F. Munagala, R.; Agrawal, A.K.; Gupta, R. New Look to Phytomedicine; Elsevier: Amsterdam, 2019, pp. 237-272.
Benarba, B.; Pandiella, A. Colorectal cancer and medicinal plants: Principle findings from recent studies. Biomed. Pharmacother., 2018, 107, 408-423.
[] [PMID: 30099345]
Aung, T.N.; Qu, Z.; Kortschak, R.D.; Adelson, D.L. Understanding the effectiveness of natural compound mixtures in cancer through their molecular mode of action. Int. J. Mol. Sci., 2017, 18(3), 656.
[] [PMID: 28304343]
Nobili, S.; Lippi, D.; Witort, E.; Donnini, M.; Bausi, L.; Mini, E.; Capaccioli, S. Natural compounds for cancer treatment and prevention. Pharmacol. Res., 2009, 59(6), 365-378.
[] [PMID: 19429468]
Banerjee, S.; Padhye, S.; Azmi, A.; Wang, Z.; Philip, P.A.; Kucuk, O.; Sarkar, F.H.; Mohammad, R.M. Review on molecular and therapeutic potential of thymoquinone in cancer. Nutr. Cancer, 2010, 62(7), 938-946.
[] [PMID: 20924969]
Yadav, R.; Bhattacharyya, A. A 745-year chronology of Cedrus deodara from western Himalaya, India. Dendrochronologia, 1992, 10, 53-61.
Chevalier, A. The encyclopedia of medicinal plant: a practical reference guide to over 550 key herbs and their medicinal uses; Dorling-Kinderseley: London, 1996, p. 227.
Chopra, R.N.; Chopra, R.N. Supplement to glossary of Indian medicinal plants; Council of Scientific & Industrial Research: New Dehli,, 1969.
Aggarwal, B.B.; Prasad, S.; Reuter, S.; Kannappan, R.; Yadev, V.R.; Park, B.; Kim, J.H.; Gupta, S.C.; Phromnoi, K.; Sundaram, C.; Prasad, S.; Chaturvedi, M.M.; Sung, B. Identification of novel anti-inflammatory agents from Ayurvedic medicine for prevention of chronic diseases: “reverse pharmacology” and “bedside to bench” approach. Curr. Drug Targets, 2011, 12(11), 1595-1653.
[] [PMID: 21561421]
Zeng, W.C.; Zhang, Z.; Gao, H.; Jia, L.R.; He, Q. Chemical composition, antioxidant, and antimicrobial activities of essential oil from pine needle (Cedrus deodara). J. Food Sci., 2012, 77(7), C824-C829.
[] [PMID: 22757704]
Chaudhary, A.K.; Ahmad, S.; Mazumder, A. Cedrus deodara (Roxb.) Loud.: a review on its ethnobotany, phytochemical and pharmacological profile. Pharmacogn. J., 2011, 3(23), 12-17.
Chung, I.M.; Lim, J.D.; Yu, B.R.; Kim, S.H.; Ateeque, A. Chemical composition of the essential oil and petroleum ether extract from Korean pine needle leaves of Cedrus deodara. Asian J. Chem., 2014, 26, 3029-3032.
Chaudhary, A.; Sharma, P.; Nadda, G.; Tewary, D.K.; Singh, B. Chemical composition and larvicidal activities of the Himalayan cedar, Cedrus deodara essential oil and its fractions against the diamondback moth, Plutella xylostella. J. Insect Sci., 2011, 11, 157-177.
[] [PMID: 22239128]
Saab, A.M.; Gambari, R.; Sacchetti, G.; Guerrini, A.; Lampronti, I.; Tacchini, M.; El Samrani, A.; Medawar, S.; Makhlouf, H.; Tannoury, M.; Abboud, J.; Diab-Assaf, M.; Kijjoa, A.; Tundis, R.; Aoun, J.; Efferth, T. Phytochemical and pharmacological properties of essential oils from Cedrus species. Nat. Prod. Res., 2018, 32(12), 1415-1427.
[] [PMID: 28670915]
Thusoo, S.; Gupta, S.; Sudan, R.; Kour, J.; Bhagat, S.; Hussain, R.; Bhagat, M. Antioxidant activity of essential oil and extracts of Valeriana jatamansi roots. BioMed Res. Int., 2014, 2014614187
Kumar, A.; Suravajhala, R.; Bhagat, M. Bioactive potential of Cedrus deodara (Roxb.) Loud essential oil (bark) against Curvularia lunata and molecular docking studies. SN Appl. Sci., 2020, 2, 1045.
Godwin, M.A.; Mahithashri, K.; Shiney, O.J.; Bhagat, M.; Praseetha, P.K. Metal incorporated g-C3N4 nanosheets as potential cytotoxic agents for promoting free radical scavenging in cancer cell lines. J. Nanosci. Nanotechnol., 2019, 19(9), 5448-55.
Hara-Chikuma, M.; Verkman, A.S. Aquaporin-3 facilitates epidermal cell migration and proliferation during wound healing. J. Mol. Med. (Berl.), 2008, 86(2), 221-231.
[] [PMID: 17968524]
Franken, N.A.; Rodermond, H.M.; Stap, J.; Haveman, J.; van Bree, C. Clonogenic assay of cells in vitro. Nat. Protoc., 2006, 1(5), 2315-2319.
[] [PMID: 17406473]
Radisky, D.C.; Levy, D.D.; Littlepage, L.E.; Liu, H.; Nelson, C.M.; Fata, J.E.; Leake, D.; Godden, E.L.; Albertson, D.G.; Nieto, M.A.; Werb, Z.; Bissell, M.J. Rac1b and reactive oxygen species mediate MMP-3-induced EMT and genomic instability. Nature, 2005, 436(7047), 123-127.
[] [PMID: 16001073]
Eruslanov, E.; Kusmartsev, S. Advanced protocols in oxidative stress II; Springer: Berlin, 2010, pp. 57-72.
Cummings, B.S.; Schnellmann, R.G. Measurement of cell death in mammalian cells. Current Protocols in Pharmacol., 2004, 25(1), 11-12.
Welinder, C.; Ekblad, L. Coomassie staining as loading control in Western blot analysis. J. Proteome Res., 2011, 10(3), 1416-1419.
[] [PMID: 21186791]
Yang, J.M. Development and evaluation of a generic evolutionary method for protein-ligand docking. J. Comput. Chem., 2004, 25(6), 843-857.
[] [PMID: 15011256]
Yang, J-M.; Shen, T-W. Proceedings of the 2004 congress on evolutionary computation, Portland: USA, 2004, 1, pp. 1028-1035.
Swamy, M.K.; Akhtar, M.S.; Sinniah, U.R. Antimicrobial properties of plant essential oils against human pathogens and their mode of action: an updated review. Evid.-. Based Complementary Altern. Med., 2016, 20163012462
Das, S.; Bordoloi, R.; Newar, N. A review on immune modulatory effect of some traditional medicinal herbs. J. Pharma. Chem. Biological Sciences, 2014, 2(1), 33-42.
Rhafouri, R.; Strani, B.; Zair, T.; Ghanmi, M.; Aafi, A.; El Omari, M.; Bentayeb, A. Chemical composition, antibacterial and antifungal activities of the Cedrus atlantica (Endl.) Manettiex Carrière seeds essential oil. Mediterranean J. Chem., 2014, 3(5), 1034-1043.
Willför, S.; Ali, M.; Karonen, M.; Reunanen, M.; Arfan, M.; Harlamow, R. Extractives in bark of different conifer species growing in Pakistan. Holzforschung, 2009, 63(5), 551-558.
Kim, Y-S.; Shin, D-H. Volatile components and antibacterial effects of pine needle (Pinus densiflora S. and Z.) extracts. Food Microbiol., 2005, 22(1), 37-45.
Tiwari, A.K.; Srinivas, P.V.; Kumar, S.P.; Rao, J.M. Free radical scavenging active components from Cedrus deodara. J. Agric. Food Chem., 2001, 49(10), 4642-4645.
[] [PMID: 11600001]
Chamorro, E.R.; Zambón, S.N.; Morales, W.G.; Sequeira, A.F.; Velasco, G.A. Study of the chemical composition of essential oils by gas chromatography. In: Gas chromatography in plant science, wine technology, toxicology and some specific applications; ; InTechOpen: London, . , 2012, pp. 307-324.
Sharma, S.H.; Thulasingam, S.; Nagarajan, S. Chemopreventive agents targeting tumor microenvironment. Life Sci., 2016, 145, 74-84.
[] [PMID: 26679106]
Hsu, S.; Singh, B.; Schuster, G. Induction of apoptosis in oral cancer cells: agents and mechanisms for potential therapy and prevention. Oral Oncol., 2004, 40(5), 461-473.
[] [PMID: 15006617]
Elmore, S. Apoptosis: a review of programmed cell death. Toxicol. Pathol., 2007, 35(4), 495-516.
[] [PMID: 17562483]
Green, D.; Kroemer, G. The central executioners of apoptosis: caspases or mitochondria? Trends Cell Biol., 1998, 8(7), 267-271.
[] [PMID: 9714597]
Kroemer, G.; Galluzzi, L.; Brenner, C. Mitochondrial membrane permeabilization in cell death. Physiol. Rev., 2007, 87(1), 99-163.
[] [PMID: 17237344]
Dröge, W. Free radicals in the physiological control of cell function. Physiol. Rev., 2002, 82(1), 47-95.
[] [PMID: 11773609]
Trachootham, D.; Alexandre, J.; Huang, P. Targeting cancer cells by ROS-mediated mechanisms: a radical therapeutic approach? Nat. Rev. Drug Discov., 2009, 8(7), 579-591.
[] [PMID: 19478820]
Hoesel, B.; Schmid, J.A. The complexity of NF-κB signaling in inflammation and cancer. Mol. Cancer, 2013, 12(1), 86.
[] [PMID: 23915189]
Pereira, S.G.; Oakley, F. Nuclear factor-kappaB1: regulation and function. Int. J. Biochem. Cell Biol., 2008, 40(8), 1425-1430.
[] [PMID: 17693123]
Zhang, X-A.; Zhang, S.; Yin, Q.; Zhang, J. Quercetin induces human colon cancer cells apoptosis by inhibiting the nuclear factor-kappa B Pathway. Pharmacogn. Mag., 2015, 11(42), 404-409.
[] [PMID: 25829782]
Moon, D-O.; Kim, M-O.; Lee, J-D.; Choi, Y.H.; Kim, G-Y. Rosmarinic acid sensitizes cell death through suppression of TNF-α-induced NF-kappaB activation and ROS generation in human leukemia U937 cells. Cancer Lett., 2010, 288(2), 183-191.
[] [PMID: 19619938]
Karin, M.; Lin, A. NF-kappaB at the crossroads of life and death. Nat. Immunol., 2002, 3(3), 221-227.
[] [PMID: 11875461]
Xia, Y.; Shen, S.; Verma, I.M. NF-κB, an active player in human cancers. Cancer Immunol. Res., 2014, 2(9), 823-830.
[] [PMID: 25187272]
Youle, R.J.; Strasser, A. The BCL-2 protein family: opposing activities that mediate cell death. Nat. Rev. Mol. Cell Biol., 2008, 9(1), 47-59.
[] [PMID: 18097445]

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