Anticancer Effect of Carob Bean Extract on Human Prostate Cancer Cell Lines: Apoptosis Induction and Bax/Bcl-2 Ratio Improvement

Khazaei, Fatemeh; Bozorgi, Maryam; Khazaei, Mozafar

doi:10.30476/mejc.2022.93823.1704

Anticancer Effect of Carob Bean Extract on Human Prostate Cancer Cell Lines: Apoptosis Induction and Bax/Bcl-2 Ratio Improvement

Document Type : Original Article(s)

Authors

Fatemeh Khazaei

Maryam Bozorgi

Mozafar Khazaei

Fertility and Infertility Research Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran

https://doi.org/10.30476/mejc.2022.93823.1704

Abstract

Background: Carob (Ceratonia siliqua L.) has been used to cure various diseases in traditional medicine. This plant has also exerted antiproliferative effects on certain cancer types. The present study aimed to determine the effects of carob bean extracts on proliferation and apoptotic genes (Bax, Bcl-2, and P53) and Caspase-3, -8, and -9 expressions of human prostate cancer cell lines.
Method: In this in vitro experimental study, human prostate cell lines (LNCaP and PC3) were treated with carob bean extract (0, 50, 100, 200, 400, and 800 μg/ml) for 24, 48, 72, and 96 h. Cell viability was investigated via MTT assay. The genes expression: Bax (pro-apoptotic), Bcl-2 (anti-apoptotic), Caspase-3 (required enzyme for the execution of apoptosis), Caspase-8 (mediator of the extrinsic pathway), and Caspase-9 (mediator of the intrinsic pathway) genes, as well as P53 (tumor suppressor), were assessed using real-time polymerase chain reaction. Moreover, the nitric oxide in culture media was evaluated.
Results: Carob bean extract suppressed the proliferation of prostate cancer cells in a time and dose-dependent manner by inducing intrinsic apoptotic pathways and decreasing nitric oxide production (P < 0.01). The obtained results revealed the overexpression of Caspases-3 and -9, P53, and Bax, but reduction in Bcl-2 expression, giving rise to a higher Bax/Bcl-2 ratio (P < 0.05).
Conclusion: The carob bean extracts exerted anti-prostate cancer properties via induction of apoptosis. It could be also suggested as a dietary supplement for patients suffering from prostate cancer.

Highlights

Fatemeh Khazaei (Google Scholar)

Mozafar Khazaei (Google Scholar)

Keywords

Prostatic neoplasm

Apoptosis

Antineoplastic agents

Carob

How to cite this article:

Khazaei F, Bozorgi M, Khazaei M. Anticancer effect of carob bean extract on human prostate cancer cell lines: apoptosis induction and Bax/Bcl-2 ratio improvement. Middle East J Cancer. 2023;14(3):363-9. doi: 10.30476/mejc.2022.93823.1704.

Yedjou CG, Mbemi AT, Noubissi F, Tchounwou SS, Tsabang N, Payton M, et al. Prostate cancer disparity, chemoprevention, and treatment by specific medicinal plants. Nutrients. 2019;11(2):336. doi: 10.3390/nu11020336.
Rabzia A, Khazaei M, Rashidi Z, Khazaei MR. Synergistic anticancer effect of paclitaxel and Noscapine on human prostate cancer cell lines. Iran J Pharm Res. 2017;16(4):1432-42.
Faramarzi A, Aghaz F, Golestan Jahromi M, Bakhtiari M, Khazaei M. Does supplementation of sperm freezing/thawing media with Ceratonia siliqua improve detrimental effect of cryopreservation on sperm parameters and chromatin quality in normozoospermic specimens? Cell Tissue Bank. 2019;20(3):403-9. doi: 10.1007/s10561-019-09779-2.
Khazaei AH, Faramarzi A, Khazaei M. Therapeutic and functional application of ceratonia siliqua: tradition remedies and new research finding. Journal of Knowledge & Health in Basic Medical Sciences 2022;17(3):18-25. doi.org/10.22100/jkh.v17i3.2862
Klenow S, Glei M, Haber B, Owen R, Pool-Zobel BL. Carob fibre compounds modulate parameters of cell growth differently in human HT29 colon adenocarcinoma cells than in LT97 colon adenoma cells. Food Chem Toxicol. 2008;46(4):1389-97. doi: 10.1016/j.fct.2007.09.003.
Custódio L, Fernandes E, Escapa AL, López-Avilés S, Fajardo A, Aligué R, et al. Antioxidant activity and in vitro inhibition of tumor cell growth by leaf extracts from the carob tree (Ceratonia siliqua). Pharm Biol. 2009;47(8):721-8.
Custódio L, Escapa AL, Fernandes E, Fajardo A, Aligué R, Alberício F, et al. In vitro cytotoxic effects and apoptosis induction by a methanol leaf extract of carob tree (Ceratonia siliqua L.). J Med Plant Res. 2011;5(10):1987-96.
Custodio L, Fernandes E, Escapa A, López-Avilés S, Fajardo A, Aligue R, et al. Antiproliferative and apoptotic activities of extracts from carob tree (Ceratonia siliqua L.) in MDA-MB-231 human breast cancer cells. Planta Medica. 2008;74(09):PA48. doi: 10.1055/s-0028-1084046.
Ghanemi FZ, Belarbi M, Fluckiger A, Nani A, Dumont A, De Rosny C, et al. Carob leaf polyphenols trigger intrinsic apoptotic pathway and induce cell cycle arrest in colon cancer cells. J Funct Foods. 2017;33:112-21. doi: 10.1016/j.jff.2017.03.032.
Khazaei F, Farzaei MH, Khazayel S, Khazaei M . Trifolium pratense extract induces apoptosis and decreases nitric oxide secretion in human umbilical vein endothelial cells, WCRJ. 2019; doi: 10.32113/wcrj_20203_1508
Khazaei S, Khademi A, Nasr Esfahani MH, Khazaei M, Nekoofar MH, Dummer PMH. Isolation and differentiation of Adipose-derived stem cells into Odontoblast-like cells: A preliminary in vitro study. Cell J. 2021;23(3):288-93. doi: 10.22074/cellj.2021.7325.
Khazaei F, Ghanbari E, Khazaei M. Protective effect of royal jelly against cyclophosphamide-induced thrombocytopenia and spleen and bone marrow damages in rats. Cell J. 2020;22(3):302-9. doi: 10.22074/cellj.2020.6703.
Dozmorov MG, Hurst RE, Culkin DJ, Kropp BP, Frank MB, Osban J, et al. Unique patterns of molecular profiling between human prostate cancer LNCaP and PC-3 cells. Prostate. 2009 1;69(10):1077-90. doi: 10.1002/pros.20960.
Crawford S. Is it time for a new paradigm for systemic cancer treatment? Lessons from a century of cancer chemotherapy. Front Pharmacol. 2013;4:68. doi: 10.3389/fphar.2013.00068.
Kim YM, Bombeck CA, Billiar TR. Nitric oxide as a bifunctional regulator of apoptosis. Circ Res. 1999;84(3):253-6. doi: 10.1161/01.res.84.3.253.
Tor YS, Yazan LS, Foo JB, Armania N, Cheah YK, Abdullah R, et al. Induction of apoptosis through oxidative stress-related pathways in MCF-7, human breast cancer cells, by ethyl acetate extract of Dillenia suffruticosa. BMC Complement Altern Med. 2014;14:55. doi: 10.1186/1472-6882-14-55.
Elmore S. Apoptosis: a review of programmed cell death. Toxicol Pathol. 2007;35(4):495-516. doi: 10.1080/01926230701320337.
Owen RW, Haubner R, Hull WE, Erben G, Spiegelhalder B, Bartsch H, et al. Isolation and structure elucidation of the major individual polyphenols in carob fibre. Food Chem Toxicol. 2003;41(12):1727-38. doi: 10.1016/s0278-6915(03)00200-x.
Papagiannopoulos M, Wollseifen HR, Mellenthin A, Haber B, Galensa R. Identification and quantification of polyphenols in carob fruits (Ceratonia siliqua L.) and derived products by HPLC-UV-ESI/MSn. J Agric Food Chem. 2004;52(12):3784-91. doi: 10.1021/jf030660y.
Lepley DM, Li B, Birt DF, Pelling JC. The chemopreventive flavonoid apigenin induces G2/M arrest in keratinocytes. Carcinogenesis. 1996;17(11):2367-75. doi: 10.1093/carcin/17.11.2367.
Kumazawa S, Taniguchi M, Suzuki Y, Shimura M, Kwon MS, Nakayama T. Antioxidant activity of polyphenols in carob pods. J Agric Food Chem. 2002;50(2):373-7. doi: 10.1021/jf010938r.
Kasai H, Fukada S, Yamaizumi Z, Sugie S, Mori H. Action of chlorogenic acid in vegetables and fruits as an inhibitor of 8-hydroxydeoxyguanosine formation in vitro and in a rat carcinogenesis model. Food Chem Toxicol. 2000;38(5):467-71. doi: 10.1016/s0278-6915(00)00014-4.