Bryonia dioica aqueous extract induces apoptosis and G2/M cell cycle arrest in MDA‑MB 231 breast cancer cells

Benarba,Bachir; Elmallah,Almahy; Pandiella,Atanasio

doi:10.3892/mmr.2019.10220

Bryonia dioica aqueous extract induces apoptosis and G2/M cell cycle arrest in MDA‑MB 231 breast cancer cells

Authors:
- Bachir Benarba
- Almahy Elmallah
- Atanasio Pandiella
View Affiliations

Affiliations: Laboratory Research on Biological Systems and Geomatics, Faculty of Nature and Life, University of Mascara, Mascara 29000, Algeria, Zoology Department, Faculty of Science Beni‑Suef University, Beni‑Suef 62511, Egypt, Instituto de Biología Molecular y Celular del Cáncer, CSIC‑CIBERONC-Universidad de Salamanca, 37001 Salamanca, Spain
Published online on: May 9, 2019 https://doi.org/10.3892/mmr.2019.10220
Pages: 73-80

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Bryonia dioica Jacq. is a climbing perennial herb with tuberous roots which is widely used in traditional medicine in Algeria for the treatment of cancer. The present study aimed to evaluate the apoptogenic activity and phytochemical composition of the aqueous extract of B. dioica roots growing in Algeria. The cytotoxic effect of B. dioica aqueous decoction against breast cancer MDA‑MB‑231 cells was evaluated by an MTT assay. Apoptosis induction was assessed by an Annexin V‑fluorescien iosthiocyanate assay. Propidium iodide staining of cell DNA was used to assess the effects on the cell cycle. In addition to UV‑Visible (UV‑vis) analysis, the major compounds of the extracts were determined using liquid chromatography‑mass spectrometric analyses. Our results showed that the B. dioica aqueous extract induced cell death in a time‑dependent manner. The highest inhibitory effect was produced at concentrations of 50 µg/ml or higher after 72 h of treatment (91.15±0.71%). Furthermore, the extract induced apoptosis of MDA‑MB‑231 cells. At 250 µg/ml, 64.61% of the treated MDA‑MB‑231 cells were apoptotic. This was accompanied by cell cycle arrest at G2/M phase. The percentage of cells in G2/M increased from 15.7% (untreated cells) to 59.13% (50 µg/ml) and 58.51% (250 µg/ml). The UV‑vis absorption spectra of B. dioica aqueous extract showed two absorption bands characteristic of the flavonol skeleton; 350‑385 nm (Band I) and 250‑280 nm (Band II). Myricetin (2,5,7,3,4,5‑pentahydroxylflavonol) was found to be the major compound in the B. dioica aqueous extract. These findings suggest that B. dioica could be considered a promising source for developing novel therapeutics against breast cancer.

View Figures

View References

1	Feng Y, Spezia M, Huang S, Yuan C, Zeng Z, Zhang L, Ji X, Liu W, Huang B, Liu B, et al: Breast cancer development and progression: Risk factors, cancer stem cells, signaling pathways, genomics, and molecular pathogenesis. Genes Dis. 5:77–106. 2018. View Article : Google Scholar : PubMed/NCBI
2	Rahman S and Zayed H: Breast cancer in the GCC countries: A focus on BRCA1/2 and non-BRCA1/2 genes. Gene. 668:73–76. 2018. View Article : Google Scholar : PubMed/NCBI
3	Bai X, Ni J, Beretov J, Graham P and Li Y: Cancer stem cell in breast cancer therapeutic resistance. Cancer Treat Rev. 69:152–163. 2018. View Article : Google Scholar : PubMed/NCBI
4	Assefa B, Glatzel G and Buchmann C: Ethnomedicinal uses of Hagenia abyssinica (Bruce) J.F. Gmel. Among rural communities of Ethiopia. J Ethnobiol Ethnomed. 6:202010. View Article : Google Scholar : PubMed/NCBI
5	Bouabdelli F, Djelloul A, Kaid-Omar Z, Semmoud A and Addou A: Antimicrobial activity of 22 plants used in urolithiasis medicine in western Algeria. Asian Pac J Trop Dis. 2 (Suppl 1):S530–S535. 2012. View Article : Google Scholar
6	Azzi R, Djaziri R and Lahfa F: Ethnopharmacological survey of medicinal plants used in the traditional treatment of diabetes mellitus in the North Western and South Western Algeria. J Med Plants Res. 6:2041–2050. 2012.
7	Benarba B, Belabid L, Righi K, Bekkar AA, Elouissi M, Khaldi A and Hammimed A: Ethnobotanical study of medicinal plants used by traditional healers in Mascara (North West of Algeria). J Ethnopharmacol. 175:626–637. 2015. View Article : Google Scholar : PubMed/NCBI
8	Benarba B: Medicinal plants used by traditional healers from South-West Algeria: An ethnobotanical study. J Intercult Ethnopharmacol. 5:320–330. 2016. View Article : Google Scholar : PubMed/NCBI
9	Benarba B: Use of medicinal plants by breast cancer patients in Algeria. EXCLI J. 14:1164–1166. 2015.PubMed/NCBI
10	Benarba B, Ambroise G, Aoues A, Meddah B and Vazquez A: Aristolochia longa aqueous extract triggers the mitochondrial pathway of apoptosis in BL41 Burkitt's lymphoma cells. Int J Green Pharm. 2012:45–49. 2012. View Article : Google Scholar
11	Benarba B, Pandiella A and Elmallah A: Anticancer activity, phytochemical screening and acute toxicity evaluation of an aqueous extract of Aristolochia longa L. Int J Pharm Phytopharmacol Res. 6:20–26. 2016. View Article : Google Scholar
12	Sallam AA, Hitotsuyanagi Y, Mansour ES, Ahmed AF, Gedara S, Fukaya H and Takeya K: Cucurbitacins from Bryonia cretica. Phytochem Lett. 3:117–121. 2010. View Article : Google Scholar
13	Nakashima S, Matsuda H, Kurume A, Oda Y, Nakamura S, Yamashita M and Yoshikawa M: Cucurbitacin E as a new inhibitor of cofilin phosphorylation in human leukemia U937 cells. Bioorg Med Chem Lett. 20:2994–2997. 2010. View Article : Google Scholar : PubMed/NCBI
14	Matsuda H, Nakashima S, Abdel-Halim OB, Morikawa T and Yoshikawa M: Cucurbitane-type triterpenes with anti-proliferative effects on U937 cells from an egyptian natural medicine, Bryonia cretica: Structures of new triterpene glycosides, bryoniaosides A and B. Chem Pharm Bull (Tokyo). 58:747–751. 2010. View Article : Google Scholar : PubMed/NCBI
15	Chekroun E, Bechiri A, Azzi R, Adida H, Benariba N and Djaziri R: Antidiabetic activity of two aqueous extracts of two cucurbitaceae: Citrullus colocynthis and Bryonia dioica. Phytothérapie. 15:57–66. 2017. View Article : Google Scholar
16	Dhouioui M, Boulila A, Jemli M, Schiets F, Casabianca H and Zina MS: Fatty acids composition and antibacterial activity of Aristolochia longa L. and Bryonia dioïca Jacq. Growing wild in tunisia. J Oleo Sci. 65:655–661. 2016. View Article : Google Scholar : PubMed/NCBI
17	Chekroun E, Benariba N, Adida H, Bechiri A, Azzi R and Djaziri R: Antioxidant activity and phytochemical screening of two Cucurbitaceae: Citrullus colocynthis fruits and Bryonia dioica roots. Asian Pac J Trop Dis. 5:632–637. 2015. View Article : Google Scholar
18	Benarba B, Meddah B and Aoues A: Bryonia dioica aqueous extract induces apoptosis through mitochondrial intrinsic pathway in BL41 Burkitt's lymphoma cells. J Ethnopharmacol. 141:510–516. 2012. View Article : Google Scholar : PubMed/NCBI
19	Díaz-Rodríguez E, Sanz E and Pandiella A: Antitumoral effect of Ocoxin, a natural compound-containing nutritional supplement, in small cell lung cancer. Int J Oncol. 53:113–123. 2018.PubMed/NCBI
20	Díaz-Rodríguez E, El-Mallah AM, Sanz E and Pandiella A: Antitumoral effect of Ocoxin in hepatocellular carcinoma. Oncol Lett. 14:1950–1958. 2017. View Article : Google Scholar : PubMed/NCBI
21	Sathish S, Janakiraman N and Johnson M: Phytochemical analysis of vitex altissima L. using UV–VIS, FTIR and GC-MS. Int J Pharm Sci Drug Res. 4:56–62. 2012.
22	Rodríguez-Gonzalo E, García-Gómez D and Carabias-Martínez R: Development and validation of a method for the detection and confirmation of biomarkers of exposure in human urine by means of restricted access material-liquid chromatography-tandem mass spectrometry. J Chromatogr A. 1217:40–48. 2010. View Article : Google Scholar : PubMed/NCBI
23	Roy S, Banerjee B and Vedasiromoni JR: Cytotoxic and apoptogenic effect of Swietenia mahagoni (L.) Jacq. leaf extract in human leukemic cell lines U937, K562 and HL-60. Environ Toxicol Pharm. 37:234–247. 2014. View Article : Google Scholar
24	Buranrat B, Mairuae N and Kanchanarach W: Cytotoxic and antimigratory effects of Cratoxy formosum extract against HepG2 liver cancer cells. Biomed Rep. 6:441–448. 2017. View Article : Google Scholar : PubMed/NCBI
25	Benarba B, Meddah B and Tir-Touil A: Response of bone resorption markers to Aristolochia longa intaked by Algerian breast cancer postmenopausal women. Adv Pharmacol Sci. 2014:1–4. 2014. View Article : Google Scholar
26	Sun X, Ma X, Li Q, Yang Y, Xu X, Sun J, Yu M, Cao K, Yang L, Yang G, et al: Anticancer effects of fisetin on mammary carcinoma cells via regulation of the PI3K/Akt/mTOR pathway: In vitro and in vivo studies. Int J Mol Med. 42:811–820. 2018.PubMed/NCBI
27	Piao XM, Gao F, Zhu JX, Wang LJ, Zhao X, Li X, Sheng MM and Zhang Y: Cucurbitacin B inhibits tumor angiogenesis by triggering the mitochondrial signaling pathway in endothelial cells. Int J Mol Med. 42:1018–1025. 2018.PubMed/NCBI
28	Sahpazidou D, Geromichalos GD, Stagos D, Apostolou A, Haroutounian SA, Tsatsakis AM, Tzanakakis GN, Hayes AW and Kouretas D: Anticarcinogenic activity of polyphenolic extracts from grape stems against breast, colon, renal and thyroid cancer cells. Toxicol Lett. 230:218–224. 2014. View Article : Google Scholar : PubMed/NCBI
29	de Lima AP, Pereira Fde C, Vilanova-Costa CA, Soares JR, Pereira LC, Porto HK, Pavanin LA, Dos Santos WB and Silveira-Lacerda Ede P: Induction of cell cycle arrest and apoptosis by ruthenium complex cis-(dichloro)tetramineruthenium(III) chloride in human lung carcinoma cells A549. Biol Trace Elem Res. 147:8–15. 2012. View Article : Google Scholar : PubMed/NCBI
30	Koo BC, Kim DH, Kim IR, Kim GC, Kwak HH and Park BS: A natural product, chios gum mastic, induces the death of HL-60 cells via apoptosis and cell cycle arrest. Int J Oral Biol. 36:13–21. 2011.
31	Hostanska K, Nisslein T, Freudenstein J, Reichling J and Saller R: Cimicifuga racemosa extract inhibits proliferation of estrogen receptor-positive and negative human breast carcinoma cell lines by induction of apoptosis. Breast Cancer Res Treat. 84:151–160. 2004. View Article : Google Scholar : PubMed/NCBI
32	Chakravarti B, Maurya R, Siddiqui JA, Bid HK, Rajendran SM, Yadav PP and Konwar R: In vitro anti-breast cancer activity of ethanolic extract of Wrightia tomentosa: Role of pro-apoptotic effects of oleanolic acid and urosolic acid. J Ethnopharmacol. 142:72–79. 2012. View Article : Google Scholar : PubMed/NCBI
33	Ferreira AK, de-Sá-Júnior PL, Pasqualoto KF, de Azevedo RA, Câmara DA, Costa AS, Figueiredo CR, Matsuo AL, Massaoka MH, Auada AV, et al: Cytotoxic effects of dillapiole on MDA-MB-231 cells involve the induction of apoptosis through the mitochondrial pathway by inducing an oxidative stress while altering the cytoskeleton network. Biochimie. 99:195–207. 2014. View Article : Google Scholar : PubMed/NCBI
34	Ye B, Li J, Li Z, Yang J, Niu T and Wang S: Anti-tumor activity and relative mechanism of ethanolic extract of Marsdenia tenacissima (Asclepiadaceae) against human hematologic neoplasm in vitro and in vivo. J Ethnopharmacol. 153:258–267. 2014. View Article : Google Scholar : PubMed/NCBI
35	Lee DH, Park KI, Park HS, Kang SR, Nagappan A, Kim JA, Kim EH, Lee WS, Hah YS, Chung HJ, et al: Flavonoids isolated from Korea citrus aurantium L. induce G2/M phase arrest and apoptosis in human gastric cancer AGS cells. J Evid Based Complementary Altern Med. 2012:1–11. 2012. View Article : Google Scholar
36	Zhang Q, Zhao XH and Wang ZJ: Flavones and flavonols exert cytotoxic effects on a human oesophageal adenocarcinoma cell line (OE33) by causing G2/M arrest and inducing apoptosis. Food Chem Toxicol. 46:2042–2053. 2008. View Article : Google Scholar : PubMed/NCBI
37	Gómez-Alonso S, Collins VJ, Vauzour D, Rodríguez-Mateos A, Corona G and Spencer JPE: Inhibition of colon adenocarcinoma cell proliferation by flavonols is linked to a G2/M cell cycle block and reduction in cyclin D1 expression. Food Chem. 130:493–500. 2012. View Article : Google Scholar
38	Youssef Moustafa AM, Khodair AI and Saleh MA: Isolation, structural elucidation of flavonoid constituents from Leptadenia pyrotechnica and evaluation of their toxicity and antitumor activity. Pharm Biol. 47:539–552. 2009. View Article : Google Scholar
39	Sisa M, Bonnet SL, Ferreira D and Van der Westhuizen JH: Photochemistry of flavonoids. Molecules. 15:5196–5245. 2010. View Article : Google Scholar : PubMed/NCBI
40	Tsimogiannis D, Samiotaki M, Panayotou G and Oreopoulou V: Characterization of flavonoid subgroups and hydroxy substitution by HPLC-MS/MS. Molecules. 12:593–606. 2007. View Article : Google Scholar : PubMed/NCBI
41	Lee HN, Shin SA, Choo GS, Kim HJ, Park YS, Kim BS, Kim SK, Cho SD, Nam JS, Choi CS, et al: Anti-inflammatory effect of quercetin and galangin in LPS-stimulated RAW264.7 macrophages and DNCB-induced atopic dermatitis animal models. Int J Mol Med. 41:888–898. 2018.PubMed/NCBI
42	Romagnolo DF and Selmin OI: Flavonoids and cancer prevention: A review of the evidence. J Nutr Gerontol Geriatr. 31:206–238. 2012. View Article : Google Scholar : PubMed/NCBI
43	Barros L, Dueñas M, Ferreira ICFR, Carvalho AM and Santos-Buelga C: Use of HPLC-DAD-ESI/MS to profile phenolic compounds in edible wild greens from Portugal. Food Chem. 127:169–173. 2011. View Article : Google Scholar
44	Marfe G, Tafani M, Indelicato M, Sinibaldi-Salimei P, Reali V, Pucci B, Fini M and Russo MA: Kaempferol induces apoptosis in two different cell lines via Akt inactivation, Bax and SIRT3 activation, and mitochondrial dysfunction. J Cell Biochem. 106:643–650. 2009. View Article : Google Scholar : PubMed/NCBI
45	Leung HW, Lin CJ, Hour MJ, Yang WH, Wang MY and Lee HZ: Kaempferol induces apoptosis in human lung non-small carcinoma cells accompanied by an induction of antioxidant enzymes. Food Chem Toxicol. 45:2005–2013. 2007. View Article : Google Scholar : PubMed/NCBI
46	Brusselmans K, Vrolix R, Verhoeven G and Swinnen JV: Induction of cancer cell apoptosis by flavonoids is associated with their ability to inhibit fatty acid synthase activity. J Biol Chem. 280:5636–5645. 2005. View Article : Google Scholar : PubMed/NCBI
47	Kang JW, Kim JH, Song K, Kim SH, Yoon JH and Kim KS: Kaempferol and quercetin, components of Ginkgo biloba extract (EGb 761), induce caspase-3-dependent apoptosis in oral cavity cancer cells. Phytother Res. 24 (Suppl 1):S77–S82. 2010. View Article : Google Scholar : PubMed/NCBI
48	Devi KP, Rajavel T, Habtemariam S, Nabavi SF and Nabavi SM: Molecular mechanisms underlying anticancer effects of myricetin. Life Sci. 142:19–25. 2015. View Article : Google Scholar : PubMed/NCBI
49	Kim ME, Ha TK, Yoon JH and Lee JS: Myricetin induces cell death of human colon cancer cells via BAX/BCL2-dependent pathway. Anticancer Res. 34:701–706. 2014.PubMed/NCBI
50	Xu Y, Xie Q, Wu S, Yi D, Yu Y, Liu S, Li S and Li Z: Myricetin induces apoptosis via endoplasmic reticulum stress and DNA double-strand breaks in human ovarian cancer cells. Mol Med Rep. 13:2094–2100. 2016. View Article : Google Scholar : PubMed/NCBI
51	Zheng AW, Chen YQ, Zhao LQ and Feng JG: Myricetin induces apoptosis and enhances chemosensitivity in ovarian cancer cells. Oncol Lett. 13:4974–4978. 2017. View Article : Google Scholar : PubMed/NCBI
52	Morales P and Haza AI: Selective apoptotic effects of piceatannol and myricetin in human cancer cells. J Appl Toxicol. 32:986–993. 2012. View Article : Google Scholar : PubMed/NCBI
53	Zhang S, Wang L, Liu H, Zhao G and Ming L: Enhancement of recombinant myricetin on the radiosensitivity of lung cancer A549 and H1299 cells. Diagn Pathol. 9:682014. View Article : Google Scholar : PubMed/NCBI
54	de Oliveira Júnior RG, Christiane Adrielly AF, da Silva Almeida JRG, Grougnet R, Thiéry V and Picot L: Sensitization of tumor cells to chemotherapy by natural products: A systematic review of preclinical data and molecular mechanisms. Fitoterapia. 129:383–400. 2018. View Article : Google Scholar : PubMed/NCBI
55	Lu J, Papp LV, Fang J, Rodriguez-Nieto S, Zhivotovsky B and Holmgren A: Inhibition of mammalian thioredoxin reductase by some flavonoids: Implications for myricetin and quercetin anticancer activity. Cancer Res. 66:4410–4418. 2006. View Article : Google Scholar : PubMed/NCBI
56	Maggioni D, Nicolini G, Rigolio R, Biffi L, Pignataro L, Gaini R and Gravello W: Myricetin and naringenin inhibit human squamous cell carcinoma proliferation and migration in vitro. Nutr Cancer. 66:1257–1267. 2014. View Article : Google Scholar : PubMed/NCBI
57	Deepa M, Sureshkumar T, Satheeshkumar PK and Priya S: Antioxidant rich Morus alba leaf extract induces apoptosis in human colon and breast cancer cells by the downregulation of nitric oxide produced by inducible nitric oxide synthase. Nutr Cancer. 65:305–310. 2013. View Article : Google Scholar : PubMed/NCBI
58	Jayakumar JK, Nirmala P, Praveen Kumar BA and Kumar AP: Evaluation of protective effect of myricetin, a bioflavonoid in dimethyl benzanthracene-induced breast cancer in female Wistar rats. South Asian J Cancer. 3:107–111. 2014. View Article : Google Scholar : PubMed/NCBI
59	Semwal DK, Semwal RB, Combrinck S and Viljoen A: Myricetin: A dietary molecule with diverse biological activities. Nutrients. 8:902016. View Article : Google Scholar : PubMed/NCBI
60	Martínez-Pérez C, Ward C, Turnbull AK, Mullen P, Cook G, Meehan J, Jarman EJ, Thomson PI, Campbell CJ, McPhail D, et al: Antitumour activity of the novel flavonoid Oncamex in preclinical breast cancer models. Br J Cancer. 114:905–916. 2016. View Article : Google Scholar : PubMed/NCBI