Anti-Tumor Effects of the Methanol Extract of Ecballium elaterium (L.) A. Rich Fruits on Human Gastric (MKN-45) and Breast (MCF-7) Cancer Cell Lines

  • Samaneh Keshavarz Medical Biotechnology Research Center, Guilan University of Medical Sciences, Rasht, Iran
  • Fatemeh Yousefbeyk Department of Pharmacognosy, School of Pharmacy, Guilan University of Medical Sciences, Rasht, Iran
  • Saghi Jani Kargar Moghaddam Medical Biotechnology Research Center, Guilan University of Medical Sciences, Rasht, Iran
  • Mahsa Ramezanpour Medical Biotechnology Research Center, Guilan University of Medical Sciences, Rasht, Iran
  • Puyan Daei Medical Biotechnology Research Center, Guilan University of Medical Sciences, Rasht, Iran
  • Saeed Ghasemi Department of Medicinal Chemistry, School of Pharmacy, Guilan University of Medical Sciences, Rasht, Iran
  • Masoud Hamidi Medical Biotechnology Research Center, Guilan University of Medical Sciences, Rasht, Iran
  • Javad Khalili Fard* Razi Herbal Medicines Research Center, Lorestan University of Medical Sciences, Khorramabad, Iran. Department of Pharmacology and Toxicology, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran https://orcid.org/0000-0002-0353-1806
DOI: https://doi.org/10.22087/hmj.v6i2.875
Keywords: Ecballium elaterium extract, Breast cancer, Apoptosis, p53, bcl-2, Bax

Abstract

Background and Aim: The Ecballium elaterium (L.) A. Rich belongs to the Cucurbitaceae family and is a Mediterranean medicinal herb with a medicinal background. In the present study, following our previous research, we aimed to evaluate the expression profile of certain significant genes involved in the process of apoptosis in human gastric (MKN-45) and breast (MCF-7) cancer cell lines treated with the methanol extract of E. elaterium fruits. Materials and Methods: The real-time PCR method was employed to calculate the expression levels of p53, bax, and bcl-2 genes in both MKN-45 and MCF-7 cell lines. Moreover, the bax/bcl-2 mRNA ratio was calculated in both cell lines. Results: Real-time PCR revealed that E. elaterium fruit extract-treated MKN-45 cells underwent a strong reduction in the mRNA levels of bcl-2 (0.04-fold) (p < 0.0001) together with an increase in p53 (18.23-fold) (p < 0.0001) and a significant reduction in bax (0.31-fold) (p <0.01). However, the up-regulation of bax (55.51-fold) (p< 0.0001) and bcl-2 (17.68-fold) (p< 0.05) genes with a concomitant revealed no significant result for p53 (2.32-fold) gene observed in MCF-7 cells. This extract had strong apoptotic activity on MKN-45 cells. The bax/bcl-2 mRNA ratio in E. elaterium fruit extract-treated MKN-45 cells was significant in comparison with the control group (7.24-fold higher and p= 0.0133), but this ratio was not remarkable in E. elaterium fruit extract-treated MCF-7 cells compared with the control group (3.95-fold higher and p= 0.0811). Conclusion: The results of this research indicated that E. elaterium fruit extract could be a promising therapeutic option against gastric cancer.

References

Touihri-Barakati I, Kallech-Ziri O, Ayadi W, Kovacic H, Hanchi B, Hosni K, et al. Cucurbitacin B purified from Ecballium elaterium (L.) A. Rich from Tunisia inhibits α5β1 integrin-mediated adhesion, migration, proliferation of human glioblastoma cell line and angiogenesis. European journal of pharmacology. 2017;797:153-61.

Ku JM, Hong SH, Kim HI, Lim YS, Lee SJ, Kim M, et al. Cucurbitacin D exhibits its anti-cancer effect in human breast cancer cells by inhibiting Stat3 and Akt signaling. European Journal of Inflammation. 2018;16:1721727X17751809.

Joulaei H, Zarei N. Women’s Cancer Care in Iran. Cancer Control. 2019;26(1):1073274819848432.

Jafargholizadeh N, Zargar SJ, Aftabi Y. The cucurbitacins D, E, and I from Ecballium elaterium (L.) upregulate the LC3 gene and induce cell-cycle arrest in human gastric cancer cell line AGS. Iranian journal of basic medical sciences. 2018;21(3):253.

Mousavi SH, Tavakkol-Afshari J, Brook A, Jafari-Anarkooli I. Role of caspases and Bax protein in saffron-induced apoptosis in MCF-7 cells. Food and chemical toxicology. 2009;47(8):1909-13.

Ju J-h, Jeon MJ, Yang W, Lee K-m, Seo H-S, Shin I. Induction of apoptotic cell death by Pharbitis nil extract in HER2-overexpressing MCF-7 cells. Journal of ethnopharmacology. 2011;133(1):126-31.

Ghoncheh M, Pournamdar Z, Salehiniya H. Incidence and mortality and epidemiology of breast cancer in the world. Asian Pacific Journal of Cancer Prevention. 2016;17(S3):43-6.

Eghdami A, Ostovar R, Jafari A, Palmer AJ, Bordbar N, Ravangard R. Economic Burden of Gastric Cancer: A Case of Iran. Cancer Control. 2019;26(1):1073274819837185.

Jafargholizadeh N, Zargar SJ, Yassa N, Tavakoli S. Purification of cucurbitacins D, E, and I from Ecballium elaterium (L.) A. rich fruits and study of their cytotoxic effects on the AGS cell line. Asian Pacific journal of cancer prevention: APJCP. 2016;17(10):4631.

Hamidi M, Ziaee M, Delashoub M, Marjani M, Karimitabar F, Khorami A, et al. The effects of essential oil of Lavandula Angustifolia on sperm parameters quality and reproductive hormones in rats exposed to cadmium. Journal of Reports in Pharmaceutical Sciences. 2015;4(2):135-42.

Elkady AI, Abuzinadah OA, Baeshen NA, Rahmy TR. Differential control of growth, apoptotic activity, and gene expression in human breast cancer cells by extracts derived from medicinal herbs Zingiber officinale. Journal of Biomedicine and Biotechnology. 2012;2012.

Safarzadeh E, Shotorbani SS, Baradaran B. Herbal medicine as inducers of apoptosis in cancer treatment. Advanced pharmaceutical bulletin. 2014;4(Suppl 1):421.

Khanaki K, Abedinzade M, Hamidi M. The effects of Urtica dioica and Lamium album extracts on the expression level of Cyclooxygenase-2 and Caspase-3 in the liver and kidney of streptozotocin-induced diabetic rats. Pharmaceutical Sciences. 2019;25(1):37-43.

Kuruppu AI, Paranagama P, Goonasekara CL. Medicinal plants commonly used against cancer in traditional medicine formulae in Sri Lanka. Saudi Pharmaceutical Journal. 2019;27(4):565-73.

Desai AG, Qazi GN, Ganju RK, El-Tamer M, Singh J, Saxena AK, et al. Medicinal plants and cancer chemoprevention. Current drug metabolism. 2008;9(7):581-91.

Chiu LC-M, Ho T-S, Wong EY-L, Ooi VE. Ethyl acetate extract of Patrinia scabiosaefolia downregulates anti-apoptotic Bcl-2/Bcl-XL expression, and induces apoptosis in human breast carcinoma MCF-7 cells independent of caspase-9 activation. Journal of ethnopharmacology. 2006;105(1-2):263-8.

Pilane M, Bagla V, Mokgotho M, Mbazima V, Matsebatlela T, Ncube I, et al. Free radical scavenging activity: antiproliferative and proteomics analyses of the differential expression of apoptotic proteins in MCF-7 cells treated with acetone leaf extract of Diospyros lycioides (Ebenaceae). Evidence-Based Complementary and Alternative Medicine. 2015;2015.

Kazemi N, Shahrestani SB. Effect of Saffron Extract on Expression of Bax and Bcl-2 Genes in Gastric Adenocarcinoma Cell Line (AGS). Gene, Cell and Tissue. 2018;5(3).

Opferman JT, Kothari A. Anti-apoptotic BCL-2 family members in development. Cell Death & Differentiation. 2018;25(1):37-45.

Liu T, Xiong J, Yi S, Zhang H, Zhou S, Gu L, et al. FKBP12 enhances sensitivity to chemotherapy-induced cancer cell apoptosis by inhibiting MDM2. Oncogene. 2017;36(12):1678-86.

Warren CF, Wong-Brown MW, Bowden NA. BCL-2 family isoforms in apoptosis and cancer. Cell death & disease. 2019;10(3):1-12.

Chang H, Li C, Huo K, Wang Q, Lu L, Zhang Q, et al. Luteolin prevents H2O2-induced apoptosis in H9C2 cells through modulating Akt-P53/Mdm2 signaling pathway. BioMed research international. 2016;2016.

Jacquot C, Rousseau B, Carbonnelle D, Chinou I, Malleter M, Tomasoni C, et al. Cucurbitacin-D-induced CDK1 mRNA up-regulation causes proliferation arrest of a non-small cell lung carcinoma cell line (NSCLC-N6). Anticancer research. 2014;34(9):4797-806.

Kianbakht S, Ziaee M, Momtaz M. A Toxicological and Phytochemical Study on the Iran’s Ecballium elaterium (L.) A. Rich. Journal of Medicinal Plants. 2019;3(71):67-76.

Bourebaba L, Gilbert-López B, Oukil N, Bedjou F. Phytochemical composition of Ecballium elaterium extracts with antioxidant and anti-inflammatory activities: Comparison among leaves, flowers and fruits extracts. Arabian Journal of Chemistry. 2020;13(1):3286-300.

Felhi S, Daoud A, Hajlaoui H, Mnafgui K, Gharsallah N, Kadri A. Solvent extraction effects on phytochemical constituents profiles, antioxidant and antimicrobial activities and functional group analysis of Ecballium elaterium seeds and peels fruits. Food Science and Technology. 2017;37(3):483-92.

Yıilmaz K, Karakuş F, Eyol E, Tosun E, Yıilmaz İ, Ünüvar S. Cytotoxic Effects of Cucurbitacin I and Ecballium elaterium on Breast Cancer Cells. Natural Product Communications. 2018;13(11):1934578X1801301108.

Bohlooli S, Jafari N, Jahed S. Cytotoxic effect of freeze-dried extract of Ecballium elaterium fruit on gastric adenocarcinoma (AGS) and esophageal squamous cell carcinoma (KYSE30) cell lines. Journal of gastrointestinal cancer. 2012;43(4):579-83.

Christodoulakis NS, Kollia K, Fasseas C. Leaf structure and histochemistry of Ecballium elaterium (L.) A. Rich.(squirting cucumber). Flora-Morphology, Distribution, Functional Ecology of Plants. 2011;206(3):191-7.

Hamidi M, Ghasemi S, Bavafa Bighdilou B, Eghbali Koohi D. Evaluation of antioxidant, antibacterial and cytotoxic activity of methanol extract from leaves and fruits of Iranian squirting cucumber (Ecballium elaterium (L.) A. Rich). Research Journal of Pharmacognosy. 2020;7(1):23-9.

Rodríguez A, Duyvejonck H, Van Belleghem JD, Gryp T, Van Simaey L, Vermeulen S, et al. Comparison of procedures for RNA-extraction from peripheral blood mononuclear cells. PloS one. 2020;15(2):e0229423.

Asai S, Ianora A, Lauritano C, Lindeque PK, Carotenuto Y. High-quality RNA extraction from copepods for Next Generation Sequencing: A comparative study. Marine genomics. 2015;24:115-8.

Zargar A, Chang S, Kothari A, Snijders AM, Mao J-H, Wang J, et al. Overcoming the challenges of cancer drug resistance through bacterial-mediated therapy. Chronic diseases and translational medicine. 2019;5(4):258-66.

Singh R, Letai A, Sarosiek K. Regulation of apoptosis in health and disease: the balancing act of BCL-2 family proteins. Nature Reviews Molecular Cell Biology. 2019;20(3):175-93.

Carneiro BA, El-Deiry WS. Targeting apoptosis in cancer therapy. Nature Reviews Clinical Oncology. 2020:1-23.

Jahanban-Esfahlan R, Seidi K, Monfaredan A, Shafie-Irannejad V, Abbasi MM, Karimian A, et al. The herbal medicine Melissa officinalis extract effects on gene expression of p53, Bcl-2, Her2, VEGF-A and hTERT in human lung, breast and prostate cancer cell lines. Gene. 2017;613:14-9.

Ekor M. The growing use of herbal medicines: issues relating to adverse reactions and challenges in monitoring safety. Frontiers in pharmacology. 2014;4:177.

Liang J, Chen D. Advances in research on the anticancer mechanism of the natural compound cucurbitacin from Cucurbitaceae plants: a review. Traditional Medicine Research. 2019;4(2):68.

Touihri I, Kallech-Ziri O, Boulila A, Fatnassi S, Marrakchi N, Luis J, et al. Ecballium elaterium (L.) A. Rich. seed oil: Chemical composition and antiproliferative effect on human colonic adenocarcinoma and fibrosarcoma cancer cell lines. Arabian Journal of Chemistry. 2019;12(8):2347-55.

Molavi O, Torkzaban F, Jafari S, Asnaashari S, Asgharian P. Chemical Compositions and Anti-Proliferative Activity of the Aerial Parts and Rhizomes of Squirting Cucumber, Cucurbitaceae. Jundishapur Journal of Natural Pharmaceutical Products. 2020;15(1).

Huang W-W, Yang J-S, Lin M-W, Chen P-Y, Chiou S-M, Chueh F-S, et al. Cucurbitacin E induces G2/M phase arrest through STAT3/p53/p21 signaling and provokes apoptosis via Fas/CD95 and mitochondria-dependent pathways in human bladder cancer T24 cells. Evidence-Based Complementary and Alternative Medicine. 2012;2012.

Zhang ZR, Gao MX, Yang K. Cucurbitacin B inhibits cell proliferation and induces apoptosis in human osteosarcoma cells via modulation of the JAK2/STAT3 and MAPK pathways. Experimental and Therapeutic Medicine. 2017;14(1):805-12.

Ma G, Luo W, Lu J, Ma D-L, Leung C-H, Wang Y, et al. Cucurbitacin E induces caspase-dependent apoptosis and protective autophagy mediated by ROS in lung cancer cells. Chemico-Biological Interactions. 2016;253:1-9.

Comşa Ş, Cimpean AM, Raica M. The story of MCF-7 breast cancer cell line: 40 years of experience in research. Anticancer research. 2015;35(6):3147-54.

Camarillo IG, Xiao F, Madhivanan S, Salameh T, Nichols M, Reece LM, et al. Low and high voltage electrochemotherapy for breast cancer: An in vitro model study. Electroporation Based Therapies Cancer. 2014:55-102.

Hotz B, Keilholz U, Fusi A, Buhr HJ, Hotz HG. In vitro and in vivo antitumor activity of cetuximab in human gastric cancer cell lines in relation to epidermal growth factor receptor (EGFR) expression and mutational phenotype. Gastric cancer. 2012;15(3):252-64.

Yokozaki H. Molecular characteristics of eight gastric cancer cell lines established in Japan. Pathology international. 2000;50(10):767-77.

Pajuelo-Lozano N, Bargiela-Iparraguirre J, Dominguez G, Quiroga AG, Perona R, Sanchez-Perez I. XPA, XPC, and XPD modulate sensitivity in gastric cisplatin resistance cancer cells. Frontiers in pharmacology. 2018;9:1197.

Alkhalaf M, El-Mowafy A. Overexpression of wild-type p53 gene renders MCF-7 breast cancer cells more sensitive to the antiproliferative effect of progesterone. Journal of endocrinology. 2003;179(1):55-62.

Oltvai Z, Millman C, Korsmeyer S. 1993. Bcl 2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell.74:609.

Yang E, Korsmeyer SJ. Molecular thanatopsis: a discourse on the BCL2 family and cell death. Blood. 1996;88(2):386-401.

Russo A, Cardile V, Graziano AC, Avola R, Bruno M, Rigano D. Involvement of Bax and Bcl-2 in induction of apoptosis by essential oils of three Lebanese Salvia species in human prostate cancer cells. International journal of molecular sciences. 2018;19(1):292.

Lin H-C, Lin J-Y. GSF3, a polysaccharide from guava (Psidium guajava L.) seeds, inhibits MCF-7 breast cancer cell growth via increasing Bax/Bcl-2 ratio or Fas mRNA expression levels. International Journal of Biological Macromolecules. 2020;161:1261-71.

Fu Z-Y, Han X-D, Wang A-H, Liu X-B. Apoptosis of human gastric carcinoma cells induced by Euphorbia esula latex. World journal of gastroenterology. 2016;22(13):3564.

Rezadoost MH, Kumleh HH, Ghasempour A. Cytotoxicity and apoptosis induction in breast cancer, skin cancer and glioblastoma cells by plant extracts. Molecular biology reports. 2019;46(5):5131-42.

Kumar S, Sharma VK, Yadav S, Dey S. Antiproliferative and apoptotic effects of black turtle bean extracts on human breast cancer cell line through extrinsic and intrinsic pathway. Chemistry Central Journal. 2017;11(1):1-10.

Patel A, Soni A, Siddiqi NJ, Sharma P. An insight into the anticancer mechanism of Tribulus terrestris extracts on human breast cancer cells. 3 Biotech. 2019;9(2):58.

Published
2023-02-12
Issue
Vol. 6 No. 2 (2021): VOL 6, NO 2, 2021
Section
Original Article

Submission of a manuscript implies: that the work described has not been published before; that it is not under consideration for publication anywhere else; that its publication has been approved by all co-authors, if any, as well as by the responsible authorities – tacitly or explicitly – at the institute where the work has been carried out. The publisher will not be held legally responsible should there be any claims for compensation.  Authors wishing to include figures, tables, or text passages that have already been published elsewhere are required to obtain permission from the copyright owner(s) for both the print and online format and to include evidence that such permission has been granted when submitting their papers. Any material received without such evidence will be assumed to originate from the authors.

This journal provides immediate open access to its content on the principle that making research freely available to the public supports a greater global exchange of knowledge. This work is licensed under a Creative Commons license (CC-BY).  However, the license permits any user to read, copy, redistribute and and make derivative the material in any medium or format for any purpose, even commercially.