Inhibition of esophageal cancer cell line (KYSE-30) proliferation using secondary metabolites of Ephedra endophyte bacteria

Ghiasvand, Mehrdad; Makhdoumi, Ali; Moghaddam Matin, Maryam; Vaezi, Jamil

doi:10.52547/nbr.8.2.95

Inhibition of esophageal cancer cell line (KYSE-30) proliferation using secondary metabolites of Ephedra endophyte bacteria

Authors

Mehrdad Ghiasvand ¹

Ali Makhdoumi ¹

Maryam Moghaddam Matin ²

Jamil Vaezi ¹

¹ Department of biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran

² Department of biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran; Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran

https://doi.org/10.52547/nbr.8.2.95

Abstract

Regarding the rising rate of esophageal cancer in some parts of Asia, including northern and northeastern regions of Iran, the identification of new anti-cancer compounds is essential to combat the disease. The relation between some plants metabolites and these endophytic microorganisms are well characterized. In the current study, the potentials of Ephedra endophyte bacteria for the inhibition of esophageal cancer cell line (KYSE-30) proliferation were investigated. A total of 54 endophyte bacteria (out of 70) were obtained from the sterilized surfaces of two medicinal plants, i.e., Ephedra intermedia and Ephedra foliata. Bacterial strains were then cultured in Tryptic soy broth (TSB) medium and, after 72 h incubation, the produced secondary metabolites were extracted by chloroform. Anticancer effects of secondary metabolites from theses bacteria on esophageal cancer cell line KYSE-30 were evaluated after 24, 48 and 72 h by MTT method. MTT assay results showed that only strain A1 had a cytotoxic effect on KYSE-30 cells. The IC₅₀amounts of this strain against KYSE-30 cell lines were equaled (µg/ml) to 346.4, 192.8 and 121.3 after 24, 48, and 72 hours, respectively. The molecular identification of strain A1 revealed that Microbacterium maritypicum (99.8% similarity) was the closest identified taxon to the strain studied. According to the promising ability of strain A1 to inhibit the growth of KYSE-30 cell line, the use of natural compounds produced by this bacterium to treat esophageal cancer was found to be applicable. However, more experiments are needed to confirm these results after purifying the ingredients, as well as conducting studies in animal cancer models.

Keywords

Actinobacter

cytotoxic effect

medicinal plant

Microbacterium

symbiosis

20.1001.1.24236330.1400.8.2.1.2

Afra, S., Makhdoumi, A., Matin, M.M. & Feizy, J. 2017. A novel red pigment from marine Arthrobacter sp. G20 with specific anticancer activity. Journal of Applied Microbiology 123: 1228-1236.
Aghababaei-Ziarati, A., Ghorbani, H. & Hosseinalizadeh, M. 2013. Evaluate the relationship between spatial distribution of esophageal and gastric cancers and soil conditions at the Golestan Province. Journal of Mazandaran University of Medical Sciences 22: 180-193. (In Persian).
Azman, A.S, Othman, I., Fang, C.M., Chan, K.G., Goh, B.H. & Lee, L.H. 2017. Antibacterial, anticancer and neuroprotective activities of rare Actinobacteria from mangrove forest soils. Indian Journal of Microbiology 57: 177-187.
Blumenthal, M. & King, P. 1995. Ma Huang: Ancient herb, modern medicine, regulatory dilemma. A review of the botany, chemistry, medicinal uses, safety concerns, and legal status of Ephedra and its alkaloids. HerbalGram 34: 22-26.
Corina, D., Delia, M., Ersilia, A., Claudia, F., Istvan, O., Andrea, B., Minda, D., Proks, M., Buda, V., Hancianu, M., Cioanca, O., Soica, C., Popescu, P. & Dehelean, C.A. 2019. Phytochemical characterization and evaluation of the antimicrobial, antiproliferative and pro-apoptotic potential of Ephedra alata Decne. Hydroalcoholic extract against the MCF-7 breast cancer cell line. Molecules 24: 13.
Cui, C.H., Kim, D.J., Jung, S.C., Kim, S.C. & Im, W.T. 2017. Enhanced production of gypenoside LXXV using a novel ginsenoside-transforming β-glucosidase from ginseng-cultivating soil bacteria and its anti-cancer property. Molecules 22: 844.
Deng, L., Ren, Z., Jia, Q., Wu, W., Shen, H. & Wang, Y. 2013. Schedule-dependent antitumor effects of 5-fluorouracil combined with sorafenib in hepatocellular carcinoma. BMC Cancer 13: 363.
Ghahremaninejad, F., Joharchi, M.R. & Vitek, E. 2005. New plant records for Khorassan province, Iran. Annalen des Naturhistorischen Museums in Wien, Serie B 106: 255-293.
Ibragic, S. & Sofić, E. 2015. Chemical composition of various Ephedra species. Bosnian Journal of Basic Medical Sciences 15: 21-27.
Karimi, N., Rashedi, J., Mahdavi Poor B., Arabi, S., Ghorbani, M., Tahmasebpour, Nahideh. & Asgharzadeh, M. 2017. Cytotoxic effects of rosemary extract on gastric adenocarcinoma (AGS) and esophageal squamous cell carcinoma (KYSE30) cell lines. Gastroenterology and Hepatology from Bed to Bench 10: 102-107.
Kumar, S., Stecher, G. & Tamura, K. 2016. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Molecular Biology and Evolution 33: 1870-1874.
Maheswarappa, G., Kavitha, D., Vijayarani, K. & Kumanan, K. 2013. Prodigiosin as anticancer drug Produced from bacteria of termite gut. Indian Journal of Basic and Applied Medical Research 1: 257-266.
Meimandi. K. & Yaghoobi, M.M. 2019. The effect of aqueous and ethanolic extracts of Sedum album L. on
human stomach and breast carcinoma cell lines in vitro. Nova Biologica Reperta 6: 10-19. (In Persian).
Napier, K.J., Scheerer, M. & Misra, S. 2014. Esophageal cancer: A review of epidemiology, pathogenesis, staging workup and treatment modalities. World Journal of Gastrointestinal Oncology 6: 112-120.
Nasiri-Madiseh, Z., Mofid, M., Ebrahimi, M., Khyam-Nekouee, S.M. & Khosravi-Ahahli, M. 2010. Isolation of taxol-producing endophytes fungi from Iranian yew (Taxus baccata L.). Journal of Shahrekord University of Medical Sciences 11: 101-106. (In Persian).
Preveena, J. & Bhore, S.J. 2013. Identification of bacterial endophytes associated with traditional medicinal plant Tridax procumbens Linn. Ancient Science of Life 32: 173-177.
Pu, X., Chen, F., Yang, Y., Qu, X., Zhang, G. & Luo, Y. 2015. Isolation and characterization of Paenibacillus polymyxa LY214, a camptothecin-producing endophytic bacterium from Camptotheca acuminata. Journal of Industrial Microbiology and Biotechnology 42: 1197-1202.
Sebola, T.E., Uche‐Okereafor, N.C., Tapfuma, K.I., Mekuto, L., Green, E. & Mavumengwana, V. 2019. Evaluating antibacterial and anticancer activity of crude extracts of bacterial endophytes from Crinum macowanii Baker bulbs. Microbiology Open 8: 914.
Seca, A. & Pinto, D. 2018. Plant secondary metabolites as anticancer agents: Successes in clinical trials and therapeutic application. International Journal of Molecular Sciences 19: 263.
Shi, Y., Liu, X., Fredimoses, M., Song, M., Chen, H., Liu, K., Lee, MH. & Dong, Z. 2018. FGFR2 regulation by picrasidine Q inhibits the cell growth and induces apoptosis in esophageal squamous cell carcinoma. Journal of Cellular Biochemistry 119: 2231-2239.
Shweta, S., Bindu, J.H., Raghu, J., Suma, H., Manjunatha, B., Kumara, P.M., Ravikanth, G., Nataraja, K.N., Ganeshaiah, K.N. & Shaanker, R.U. 2013. Isolation of endophytic bacteria producing the anti-cancer alkaloid camptothecine from Miquelia dentata Bedd. (Icacinaceae). Phytomedicine 20: 913-917.
Smyth, E.C., Lagergren, J., Fitzgerald, R.C., Lordick, F., Shah, M.A., Lagergren, P. & Cunninghamet, D. 2017. Oesophageal cancer. Nature Reviews Disease Primers 3: 17048.
Yang, L., Wen, K.S., Ruan, X., Zhao, Y.X., Wei, F. & Wang, Q. 2018. Response of plant secondary metabolites to environmental factors. Molecules 23: 762.
Zinniel, D.K., Lambrecht, P., Harris, N.B., Feng, Z., Kuczmarski, D. & Higley, P., Ishimaru, C.A., Arunakumari, A., Barletta, R.G. Vidaver, A.K. 2002. Isolation and characterization of endophytic colonizing bacteria from agronomic crops and prairie plants. Applied and Environmental Microbiology 68: 2198-2208.