Evaluation of the Cytotoxic Activity of Anthriscus nemorosa on Breast Cancer Cells
Keywords:Anthriscus nemorosa, cancer, GC-MS, Cytotoxic, Fow cytometry.
Objective: The present study concerns the cytotoxic activity of A. nemorosa different extracts on breast cancer cells (MCF-7) and normal cell lines (HFFF).
Methods: Different extracts of aerial parts of A. nemorosa were prepared using Soxhlet apparatus. The cytotoxicity of samples was assessed by MTT assay on breast cancer cells (MCF-7) and noncancerous cells (HFFF) with different concentrations of extracts in 24 and 48 hours. The most potent extract was fractioned and cytotoxic activity of fractions was considered, As well. A flow cytometry (annexin V/PI) assay has been used for detecting the mechanism of cell death in sample treated cell lines. Moreover, for clarifying volatile components of n-Hexane extract and its 80% and 100% VLC fractions were subjected to GC-MS apparatus.
Results: Results indicated that n-Hexane extract and its 80% and 100% VLC fractions exhibited a significant (p<0.001) inhibitory effect on the growth of the MCF-7 cell line compared to the control group. Meanwhile, flow cytometry analysis revealed that potent extract caused cell death through necrosis and 80% and 100% fractions showed different mechanisms (such as autophagy). The major compounds, which maybe were in charge of showing cytotoxic activity were non-terpenoids.
Conclusion: This study provides the evidence that in vitro cytotoxic activity of n-Hexane extract and 80% and 100% VLC fractions of A. nemorosa inhibited the proliferation of breast cancer cells (MCF7) via a different mechanism.
2. Zeinomar N, Knight JA, Genkinger JM, Phillips K-A, Daly MB, Milne RL, et al. Alcohol consumption, cigarette smoking, and familial breast cancer risk: findings from the Prospective Family Study Cohort (ProF-SC). Breast Cancer Research. 2019;21(1):1-14.
3. Trichopoulos D, MacMahon B, Cole P. Menopause and breast cancer risk. Journal of the National Cancer Institute. 1972;48(3):605-13.
4. Taraphdar AK, Roy M, Bhattacharya R. Natural products as inducers of apoptosis: Implication for cancer therapy and prevention. Current science. 2001:1387-96.
5. Harvey AL. Natural products in drug discovery. Drug discovery today. 2008;13(19-20):894-901.
6. Seyfi R, Kahaki FA, Ebrahimi T, Montazersaheb S, Eyvazi S, Babaeipour V, et al. Antimicrobial peptides (AMPs): roles, functions and mechanism of action. International Journal of Peptide Research and Therapeutics. 2020;26(3):1451-63.
7. Alipour M, Khanmohammadi O. Antibacterial activity of plant extracts against oral and skin pathogens. African Journal of Microbiology Research. 2011;5(19):2909-11.
8. Fulda S, Efferth T. Selected secondary plant metabolites for cancer therapy. World Journal of Traditional Chinese Medicine. 2015;1(1):24.
9. Mohammadzadeh F, Monirifar H, Saba J, Valizadeh M, Haghighi AR, Zanjani BM, et al. Genetic variation among Iranian alfalfa (Medicago sativa L.) populations based on RAPD markers. Bangladesh Journal of Plant Taxonomy. 2011;18(2):93-104.
10. Safarzadeh E, Shotorbani SS, Baradaran B. Herbal medicine as inducers of apoptosis in cancer treatment. Advanced pharmaceutical bulletin. 2014;4(Suppl 1):421.
11. Elujoba AA, Odeleye O, Ogunyemi C. Traditional medicine development for medical and dental primary health care delivery system in Africa. African Journal of Traditional, Complementary and Alternative Medicines. 2005;2(1):46-61.
12. Maikhuri R, Nautiyal S, Rao K, Saxena K. Role of medicinal plants in the traditional health care system: a case study from Nanda Devi Biosphere Reserve. Current Science. 1998:152-7.
13. Jeong G-S, Kwon O-K, Park B-Y, Oh S-R, Ahn K-S, Chang M-J, et al. Lignans and coumarins from the roots of Anthriscus sylvestris and their increase of caspase-3 activity in HL-60 cells. Biological and Pharmaceutical Bulletin. 2007;30(7):1340-3.
14. Lee SH, Son MJ, Ju HK, Lin CX, Moon TC, Choi H-G, et al. Dual inhibition of cyclooxygenases-2 and 5-lipoxygenase by deoxypodophyllotoxin in mouse bone marrow-derived mast cells. Biological and Pharmaceutical Bulletin. 2004;27(6):786-8.
15. Sudo K, Konno K, Shigeta S, Yokota T. Inhibitory effects of podophyllotoxin derivatives on herpes simplex virus replication. Antiviral Chemistry and Chemotherapy. 1998;9(3):263-7.
16. Lin CX, Lee E, Jin MH, Yook J, Quan Z, Ha K, et al. Deoxypodophyllotoxin (DPT) inhibits eosinophil recruitment into the airway and Th2 cytokine expression in an OVA-induced lung inflammation. Planta medica. 2006;72(09):786-91.
17. Kozawa M, Baba K, Matsuyama Y, Kido T, Sakai M, TAKEMOTO T. Components of the root of Anthriscus sylvestris HOFFM. II. Insecticidal activity. Chemical and Pharmaceutical Bulletin. 1982;30(8):2885-8.
18. Jin M, Moon TC, Quan Z, Lee E, Kim YK, Yang JH, et al. The naturally occurring flavolignan, deoxypodophyllotoxin, inhibits lipopolysaccharide-induced iNOS expression through the NF-ÎºB activation in RAW264. 7 macrophage cells. Biological and Pharmaceutical Bulletin. 2008;31(7):1312-5.
19. Lin CX, Son MJ, Ju HK, Moon TC, Lee E, Kim SH, et al. Deoxypodophyllotoxin, a naturally occurring lignan, inhibits the passive cutaneous anaphylaxis reaction. Planta medica. 2004;70(05):474-6.
20. KISO Y, KONNO C, HIKINO H, YAGI Y, HASHIMOTO I. Liver-protective actions of desoxypodophyllotoxin and its analogs. Journal of pharmacobio-dynamics. 1982;5(8):638-41.
21. Lee SK, Kim Y, Jin C, Lee SH, Kang MJ, Jeong TC, et al. Inhibitory effects of deoxypodophyllotoxin from Anthriscus sylvestris on human CYP2C9 and CYP3A4. Planta medica. 2010;76(07):701-4.
22. Choi H, Lee J, Shin H-J, Lee B-G, Chang I, Hwang J-S. Deoxypodophyllotoxin reduces skin pigmentation of brown guinea pigs. Planta medica. 2004;70(04):378-80.
23. Chen H, Jiang H-Z, Li Y-C, Wei G-Q, Geng Y, Ma C-Y. Antitumor constituents from Anthriscus sylvestris (L.) Hoffm. Asian Pacific Journal of Cancer Prevention. 2014;15(6):2803-7.
24. Beheshtirouy S, Mirzaei F, Eyvazi S, Tarhriz V. Recent Advances on Therapeutic Peptides for Breast Cancer Treatment. Current Protein & Peptide Science. 2020.
25. Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: a cancer journal for clinicians. 2018;68(6):394-424.
26. Asadi-Samani M, Kooti W, Aslani E, Shirzad H. A systematic review of Iranâ€™s medicinal plants with anticancer effects. Journal of evidence-based complementary & alternative medicine. 2016;21(2):143-53.
27. Lai P, Rao H, Gao Y. Chemical composition, cytotoxic, antimicrobial and antioxidant activities of essential oil from Anthriscus caucalis M. Bieb grown in China. Records of Natural Products. 2018;12(3):290-4.
28. Yong Y, Shin SY, Lee YH, Lim Y. Antitumor activity of deoxypodophyllotoxin isolated from Anthriscus sylvestris: Induction of G2/M cell cycle arrest and caspase-dependent apoptosis. Bioorganic & medicinal chemistry letters. 2009;19(15):4367-71.
29. Olaru OT, NiÅ£ulescu GM, OrÈ›an A, Dinu-PÃ®rvu CE. Ethnomedicinal, phytochemical and pharmacological profile of Anthriscus sylvestris as an alternative source for anticancer lignans. Molecules. 2015;20(8):15003-22.
30. Jung CH, Kim H, Ahn J, Jung SK, Um MY, Son K-H, et al. Anthricin isolated from Anthriscus sylvestris (L.) Hoffm. inhibits the growth of breast cancer cells by inhibiting Akt/mTOR signaling, and its apoptotic effects are enhanced by autophagy inhibition. Evidence-Based Complementary and Alternative Medicine. 2013;2013.
31. Harada H, Yamashita U, Kurihara H, Fukushi E, Kawabata J, Kamei Y. Antitumor activity of palmitic acid found as a selective cytotoxic substance in a marine red alga. Anticancer research. 2002;22(5):2587-90.
32. Kim SJ, Chung WS, Kim SS, Ko SG, Um JY. Antiinflammatory effect of Oldenlandia diffusa and its constituent, hentriacontane, through suppression of caspaseâ€1 activation in mouse peritoneal macrophages. Phytotherapy Research. 2011;25(10):1537-46.
33. Yang J-H, Lee C-H, Monteiro-Riviere NA, Riviere JE, Tsang C-L, Chou C-C. Toxicity of jet fuel aliphatic and aromatic hydrocarbon mixtures on human epidermal keratinocytes: evaluation based on in vitro cytotoxicity and interleukin-8 release. Archives of toxicology. 2006;80(8):508-23.
34. Fukuzawa M, Yamaguchi R, Hide I, Chen Z, Hirai Y, Sugimoto A, et al. Possible involvement of long chain fatty acids in the spores of Ganoderma lucidum (Reishi Houshi) to its anti-tumor activity. Biological and Pharmaceutical Bulletin. 2008;31(10):1933-7.
35. Van Uden W, Bos JA, Boeke GM, Woerdenbag HJ, Pras N. The large-scale isolation of deoxypodophyllotoxin from rhizomes of Anthriscus sylvestris followed by its bioconversion into 5-methoxypodophyllotoxin Î²-D-glucoside by cell cultures of Linum flavum. Journal of Natural Products. 1997;60(4):401-3.
36. Lv M, Xu H. Recent advances in semisynthesis, biosynthesis, biological activities, mode of action, and structure-activity relationship of podophyllotoxins: an update (2008-2010). Mini reviews in medicinal chemistry. 2011;11(10):901-9.