In collaboration with Payame Noor University and Iranian Society of Physiology and Pharmacology

Document Type : Article

Authors

1 M.A., Department of Chemistry, Payame Noor University, ‎Tehran, Iran

2 Associate professor, Department of Biology, Payame Noor ‎University, Tehran, Iran

3 Professor, Department of Chemistry, Payame Noor University, ‎Tehran, Iran

Abstract

In this research, a comparison was made between the concentration of secondary metabolites of Hibiscus sabdariffa content in its leaves and induced transgenic hairy roots. The levels of anthocyanins and flavonoids and total phenol were measured in both samples. In the next step, polyphenols were measured by HPLC. The concentration of catechin flavonoids in the transgenic hairy root extract was 22.8 mg/g dry matter which was several times higher than that of ​​0.9 mg/g in leaf extract of dry matter. The obtained results clearly demonstrated that, the rate of concentration of secondary metabolites during the process of induction of transgenic hairy root was increased. Then, the extracts of leaf and transgenic hairy root samples at various concentrations of 0, 25, 50, 50, 100, 200, 400 mg / ml were added to HepG2 liver cancer cell lines and the percentage of cell survival was calculated by MTT method. The cell survival rate in the sample treated with transgenic hairy roots samples was less than the leaf extract samples, which was most likely due to the higher concentration of Catechin in the transgenic hairy roots extract. The best concentrations of extracts of transgenic hairy root extract showed that, the highest percentage of cell apoptosis was 100 mg/ ml.

Keywords

Adhami, V.M.; Malik, A.; Zaman, N.; Sarfaraz, S.; Siddiqui, I.A.; Syed, D.N.; Afaq, F.; Pasha, F.S.; Saleem, M.; Mukhtar, H. (2007). Combined inhibitory effects of green tea polyphenols and selective cyclooxygenase-2 inhibitors on the growth of human prostate cancer cells both in vitro and in vivo, Clinical Cancer Research, 13: 1611-1619.
Ahmad, N.; Gupta, S.; Mukhtar, H. (2000). Green tea polyphenol epigallocatechin-3-gallate differentially modulates nuclear factor κB in cancer cells versus normal cells, Archives of biochemistry and biophysics, 376: 338-346.
Bettuzzi, S.; Brausi, M.; Rizzi, F.; Castagnetti, G.; Peracchia, G.; Corti, A. (2006).Chemoprevention of human prostate cancer by oral administration of green tea catechins in volunteers with high-grade prostate intraepithelial neoplasia: a preliminary report from a one-year proof-of-principle study, Cancer Res., 66: 1234-1240.
Cheng, Z.; Zhang, Z.; Han, Y.; Wang, J.; Wang, Y.; Chen, X.; Shao, Y.; Cheng, Y.; Zhou, W.; Lu, X. (2020).  A review on anti-cancer effect of green tea catechins, Journal of Functional Foods, 74: 104172.
Crespy, V.; Williamson, G. (2004).  A review of the health effects of green tea catechins in in vivo animal models, The Journal of nutrition, 134: 3431S-3440S.
Fuleki, T.; Francis, F. (1968).  Quantitative methods for anthocyanins. 2. Determination of total anthocyanin and degradation index for cranberry juice, Journal of food science, 33: 78-83.
Hanafy, M.; Matter, M.; Asker, M.; Rady, M. (2016). Production of indole alkaloids in hairy root cultures of Catharanthus roseus L. and their antimicrobial activity, South African Journal of Botany, 105: 9-18.
Higdon, J.V.; Frei, B. (2003). Tea catechins and polyphenols: health effects, metabolism, and antioxidant functions.
Huang, C.-Y.; Han, Z.; Li, H.; Xie, H.; Zhu, S. (2017).  Mechanism of EGCG promoting apoptosis of MCF-7 cell line in human breast cancer, Oncology letters, 14: 3623-3627.
Ismail, A.; Ikram, E.H.K.; Nazri, H.S.M.; (2008). Roselle (Hibiscus sabdariffa L.) seeds-nutritional composition, protein quality and health benefits, Food, 2: 1-16.
Kaszkin, M.; Beck, K.-F.; Eberhardt, W.; Pfeilschifter, J. (2004). Unravelling green tea's mechanisms of action: more than meets the eye, Molecular pharmacology, 65: 15-17.
Khan, N.; Adhami, V.M.; Mukhtar, H. (2009). Green tea polyphenols in chemoprevention of prostate cancer: preclinical and clinical studies, Nutrition and cancer, 61: 836-841.
Krumov, N.; Perner‐Nochta, I.; Oder, S.; Gotcheva, V.; Angelov, A.; Posten, C. (2009).  Production of inorganic nanoparticles by microorganisms, Chemical Engineering & Technology: Industrial Chemistry‐Plant Equipment‐Process Engineering‐Biotechnology, 32: 1026-1035.
Kumazoe, M.; Sugihara, K.; Tsukamoto, S.; Huang, Y.; Tsurudome, Y.; Suzuki, T.; Suemasu, Y.; Ueda, N.; Yamashita, S.; Kim, Y. (2013). 67-kDa laminin receptor increases cGMP to induce cancer-selective apoptosis, The Journal of clinical investigation, 123.
Kundu, S.; Salma, U.; Ali, M.N.; Hazra, A.K.; Mandal, N. (2018). Development of transgenic hairy roots and augmentation of secondary metabolites by precursor feeding in Sphagneticola calendulacea (L.) Pruski, Industrial Crops and Products, 121: 206-215.
Lambert, J.D.; Elias, R.J. (2010).  The antioxidant and pro-oxidant activities of green tea polyphenols: a role in cancer prevention, Archives of biochemistry and biophysics, 501: 65-72.
Lee, I.P.; Kim, Y.H.; Kang, M.H.; Roberts, C.; Shim, J.S.; Roh, J.K. (1997). Chemopreventive effect of green tea (Camellia sinensis) against cigarette smoke‐induced mutations (SCE) in humans, Journal of Cellular Biochemistry, 67: 68-75.
Li, H.C.; Yashiki, S.; Sonoda, J.; Lou, H.; Ghosh, S.K.; Byrnes, J.J.; Lema, C.; Fujiyoshi, T.; Karasuyama, M.; Sonoda, S. (2000).  Green tea polyphenols induce apoptosis in vitro in peripheral blood T lymphocytes of adult T‐cell leukemia patients, Japanese journal of cancer research, 91: 34-40.
Manikandan, R.; Beulaja, M.; Arulvasu, C.; Sellamuthu, S.; Dinesh, D.; Prabhu, D.; Babu, G.; Vaseeharan, B.; Prabhu, N. (2012). Synergistic anticancer activity of curcumin and catechin: An in vitro study using human cancer cell lines, Microscopy research and technique, 75: 112-116.
Mckay, D.; (2009).  Can hibiscus tea lower blood pressure, AfroFood Industry Hi-Tech, 20: 40-42.
Mitra, S.; Dash, R. (2018). Natural products for the management and prevention of breast cancer, Evidence-Based Complementary and Alternative Medicine, 2018.
Mudgal, V.; Madaan, N.; Mudgal, A.;  Mishra, S. (2010).  Dietary polyphenols and human health, Asian Journal of Biochemistry, 5: 154-162.
Nishikawa, T.; Nakajima, T.; Moriguchi, M.; Jo, M.; Sekoguchi, S.; Ishii, M.; Takashima, H.; Katagishi, T.; Kimura, H.; Minami, M. (2006). A green tea polyphenol, epigalocatechin-3-gallate, induces apoptosis of human hepatocellular carcinoma, possibly through inhibition of Bcl-2 family proteins, Journal of hepatology, 44: 1074-1082.
Oksman-Caldentey, K.-M.; Inzé, D. (2004). Plant cell factories in the post-genomic era: new ways to produce designer secondary metabolites, Trends in plant science; 9: 433-440.
Omid, M.; Farzin, N. (2013). Biotechnology solutions to increase the effectiveness of medicinal plants. New genetics; 3: 220-209.
Shankar, E.; Kanwal, R.; Candamo, M.; Gupta, S. (2016) Dietary phytochemicals as epigenetic modifiers in cancer: promise and challenges, in: Seminars in cancer biology, Elsevier, , pp. 82-99.
Shimizu, M.; Fukutomi, Y.; Ninomiya, M.; Nagura, K.; Kato, T.; Araki, H.; Suganuma, M.; Fujiki, H.; Moriwaki, H. (2008). Green tea extracts for the prevention of metachronous colorectal adenomas: a pilot study, Cancer Epidemiology and Prevention Biomarkers, 17: 3020-3025.
Shimizu, M.; Shirakami, Y.; Sakai, H.; Kubota, M.; Kochi, T.; Ideta, T.; Miyazaki, T.; Moriwaki, H. (2015).  Chemopreventive potential of green tea catechins in hepatocellular carcinoma, International Journal of Molecular Sciences, 16: 6124-6139.
Sonoda, J.I.; Ikeda, R.; Baba, Y.; Narumi, K.; Kawachi, A.; Tomishige, E.; Nishihara, K.; Takeda, Y.; Yamada, K.; Sato, K. (2014). Green tea catechin, epigallocatechin‑3‑gallate, attenuates the cell viability of human non‑small‑cell lung cancer A549 cells via reducing Bcl‑xL expression, Experimental and Therapeutic Medicine, 8: 59-63.
Srivastava, S.; Srivastava, A.K. (2007).  Hairy root culture for mass-production of high-value secondary metabolites, Critical Reviews in Biotechnology, 27: 29-43.
Suganuma, M.; Saha, A.; Fujiki, H. (2011).  New cancer treatment strategy using combination of green tea catechins and anticancer drugs, Cancer science, 102: 317-323.
Tripathi, L.; Tripathi, J.N. (2003). Role of biotechnology in medicinal plants, Tropical Journal of Pharmaceutical Research; 2: 243-253.
Wang, M.L.; Morris, B.; Tonnis, B.; Davis, J.; Pederson, G.A. (2012). Assessment of oil content and fatty acid composition variability in two economically important Hibiscus species, Journal of agricultural and food chemistry; 60: 6620-6626.
Xiang, L. P.; Wang, A.; Ye, J. H.; Zheng, X. Q.; Polito, C. A.; Lu, J. L.; Liang, Y. R. (2016).  Suppressive effects of tea catechins on breast cancer. Nutrients; 8: 1-15.
Yamauchi, R.; Sasaki, K.; Yoshida, K. (2009). Identification          of epigallocatechin-3-gallate in green tea polyphenols as a potent inducer of p53-dependent apoptosis in the human lung cancer cell line A549, Toxicology in Vitro; 23: 834-839.
Yang, C.S.; Wang, H.; Li, G.X.; Yang, Z.;  Guan, F.; Jin, H. (2011).  Cancer prevention by tea: evidence from laboratory studies, Pharmacological Research; 64: 113-122.
Yang, C.S.; Wang, X.; Lu, G.; Picinich, S.C. (2009). Cancer prevention by tea: animal studies, molecular mechanisms and human relevance, Nature Reviews Cancer; 9: 429-439.
Zhishen, J.; Mengcheng, T.; Jianming, W. (1999).  The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food chemistry; 64: 555-559.