Veterinary Research Forum
Vet Res Forum

Hydroalcoholic extract of Taraxacum officinale induces apoptosis and autophagy in 4T1 breast cancer cells

Document Type : Original Article

Authors

Sharareh Ahmadi 1
Adel Saberivand 1
Cyrus Jalili 2
Reza Asadpour 1
Monire Khordadmehr 3
Maryam Saberivand 4

1 Department of Clinical Science, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran

2 Medical Biology Research Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran

3 Department of Pathobiology, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran

4 Connective Tissue Diseases Research Center, Tabriz University of Medical Science, Tabriz, Iran

https://doi.org/10.30466/vrf.2023.1985987.3726
Abstract
Triple-negative breast cancer (TNBC) is an aggressive and deadly breast cancer sub-type with limited therapeutic options. Dandelion (Taraxacum officinale) exhibiting extensive anti-cancer activity is reported to be effective against TNBC; however, its anti-tumor effect mechanisms have not been fully elucidated. The purpose of this study was to determine the anti-cancer activity of hydroalcoholic extract of dandelion (HADE) on 4T1 cells, and the mechanism of HADE-induced cell death. The effect of HADE on cell viability was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and lactate dehydrogenase assays. Apoptotic cell death was monitored by flow cytometry. The DNA fragmentation was evaluated by Acridine orange/Ethidium bromide (AO/EB) staining. Nitric oxide (NO) level was detected using Griess assay. The effects of HADE on Atg-7, Beclin-1, Bcl2, Bax and p53 genes were investigated by real-time reverse transcription-polymerase chain reaction. The results showed that HADE inhibited cell growth and proliferation in a dose- and time-dependent manner. The HADE induced 4T1 breast cancer cell death via apoptosis and autophagy. The DNA fragmentation was improved as the concentration of HADE increased. The NO secretion was declined with increasing concentration of HADE. Gene expression analysis confirmed HADE-induced apoptosis and autophagy in cancer cells. The Bax, Bax/Bcl-2 ratio, p53, Beclin-1 and Atg-7 over-expression as well as Bcl-2 down-regulation were also evident in treated cancer cells.

Keywords

Apoptosis
Autophagy
Breast cancer
Dandelion
Taraxacum officinale

Subjects

  1. Yu K, Tan L, Lin L, et al. Deep-learning-empowered breast cancer auxiliary diagnosis for 5GB remote E-health. IEEE Wirel Commun 2021; 28(3): 54-61.
  2. Sung H, Ferlay J, Siegel RL, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 2021; 71(3): 209-249.
  3. Soerjomataram I, Bray F. Planning for tomorrow: global cancer incidence and the role of prevention 2020-2070. Nat Rev Clin Oncol 2021; 18(10): 663-672.
  4. Dai X, Li T, Bai Z, et al. Breast cancer intrinsic subtype classification, clinical use and future trends. Am J Cancer Res 2015; 5(10): 2929-2943.
  5. Kumar P, Aggarwal R. An overview of triple-negative breast cancer. Arch Gynecol Obstet 2016; 293(2): 247-269.
  6. Medina MA, Oza G, Sharma A, et al. Triple-negative breast cancer: a review of conventional and advanced therapeutic strategies. Int J Environ Res Public Halth 2020; 17(6): 2078. doi: 10.3390/ijerph17062078.
  7. Abu-Darwish MS, Efferth T. Medicinal plants from near east for cancer therapy. Front Pharmacol 2018; 9: 56. doi: 10.3389/fphar.2018.00056.
  8. Lis B, Olas B. Pro-health activity of dandelion (Taraxacum officinale) and its food products - history and present. J Funct Foods 2019; 59: 40-48.
  9. Martinez M, Poirrier P, Chamy R, et al. Taraxacum officinale and related species - an ethno-pharmacological review and its potential as a commercial medicinal plant. J Ethnopharmacol 2015; 169: 244-262.
  10. Qu J, Ke F, Liu Z, et al. Uncovering the mechanisms of dandelion against triple-negative breast cancer using a combined network pharmacology, molecular pharmacology and metabolomics approach. Phytomedicine 2022; 99: 153986. doi: 10.1016/ j.phymed.2022.153986.
  11. Ren F, Li J, Yuan X, et al. Dandelion polysaccharides exert anticancer effect on hepatocellular carcinoma by inhibiting PI3K/AKT/mTOR pathway and enhancing immune response. J Funct Foods 2019; 55: 263-274.
  12. Ovadje P, Chochkeh M, Akbari-Asl P, et al. Selective induction of apoptosis and autophagy through treatment with dandelion root extract in human pancreatic cancer cells. Pancreas 2012; 41(7):
    1039-1047.
  13. El-Emam SZ, El-Ella DMA, Fayez SM, et al. Novel dandelion mannan-lipid nanoparticle: exploring the molecular mechanism underlying the potent anticancer effect against non-small lung carcinoma. J Funct Foods 2021; 87: 104781. doi: 10.1016/ j.jff.2021.104781.
  14. Ovadje P, Ammar S, Guerrero JA, et al. Dandelion root extract affects colorectal cancer proliferation and survival through the activation of multiple death signalling pathways. Oncotarget 2016; 7(45): 73080-73100.
  15. Choi EJ, Kim GH. Dandelion (Taraxacum officinale) flower ethanol extract inhibits cell proliferation and induces apoptosis in human ovarian cancer SK-OV-3 cells. Food Sci Biotechnol 2009; 18(2): 552-555.
  16. Ovadje P, Hamm C, Pandey S. Efficient induction of extrinsic cell death by dandelion root extract in human chronic myelomonocytic leukemia (CMML) cells. PLoS One 2012; 7(2): e30604. doi: 10.1371/journal. pone.0030604.
  17. Deng XX, Jiao YN, Hao HF, et al. Taraxacum mongolicum extract inhibited malignant phenotype of triple-negative breast cancer cells in tumor-associated macrophages microenvironment through suppressing IL-10/STAT3/PD-L1 signaling pathways. J Ethno-pharmacol 2021; 274: 113978. doi: 10.1016/ j.jep.2021.113978.
  18. Li XH, He XR, Zhou YY, et al. Taraxacum mongolicum extract induced endoplasmic reticulum stress associated-apoptosis in triple-negative breast cancer cells. J Ethnopharmacol 2017; 206: 55-64.
  19. Kazemi MH, Shokrollahi Barough M, Ghanavatinejad A, et al. Decrease of tumor-infiltrating regulatory T cells using pentoxifylline: an ex vivo analysis in triple-negative breast cancer mouse model. Iran J Allergy Asthma Immunol 2022; 21(2): 167-177.
  20. Kumar P, Nagarajan A, Uchil PD. Analysis of cell viability by the MTT assay. Cold Spring Harb Protoc 2018; 6: 095505. doi: 10.1101/pdb.prot095505.
  21. Kaja S, Payne AJ, Naumchuk Y, et al. Quantification of lactate dehydrogenase for cell viability testing using cell lines and primary cultured astrocytes. Curr Protoc Toxicol 2017; 72: 2.26.1-2.26.10. doi: 10.1002/cptx.21.
  22. Wlodkowic D, Skommer J, Darzynkiewicz Z. Flow cytometry-based apoptosis detection. Methods Mol Biol 2009; 559: 19-32.
  23. Gercel-Taylor C. Diphenylamine assay of DNA fragmentation for chemosensitivity testing. Methods Mol Med 2005; 111: 79-82.
  24. Murugan S, Amaravadi RK. Methods for studying auto-phagy within the tumor microenvironment. Adv Exp Med Biol 2016; 899: 145-66.
  25. Ghasemi A, Hedayati M, Biabani H. Protein precipitation methods evaluated for determination of serum nitric oxide end products by the Griess assay. J Med Sci Res 2007; 1(1): 43-46.
  26. D'Arcy MS. Cell death: a review of the major forms of apoptosis, necrosis and autophagy. Cell Biol Int 2019; 43(6): 582-592.
  27. Fesik SW. Promoting apoptosis as a strategy for cancer drug discovery. Nat Rev Cancer 2005; 5(11): 876-885.
  28. Yun CW, Lee SH. The roles of autophagy in cancer. Int J Mol Sci 2018; 19(11): 3466. doi: 10.3390/ ijms19113466.
  29. Gewirtz DA. The four faces of autophagy: implications for cancer therapy. Cancer Res 2014; 74(3): 647-651.
  30. Wang T, Hu J, Luo H, et al. Photosensitizer and autophagy promoter coloaded ROS-responsive dendrimer-assembled carrier for synergistic enhancement of tumor growth suppression. Small 2018; 14: 1802337. doi:10.1002/smll.20180233732.
  31. Clarke PG, 31. Puyal J. Autophagic cell death exists. Autophagy 2012; 8(6): 867-869.
  32. Luo T, Fu J, Xu A, et al. PSMD10/gankyrin induces autophagy to promote tumor progression through cytoplasmic interaction with ATG7 and nuclear transactivation of ATG7 expression. Autophagy 2016; 12(8): 1355-1371.
  33. Yue Z, Jin S, Yang C, et al. Beclin 1, an autophagy gene essential for early embryonic development, is a haploin sufficient tumor suppressor. Proc Natl Acad Sci U S A 2003; 100(25): 15077-15082.
  34. Liu J, Xia H, Kim M, et al. Beclin1 controls the levels of p53 by regulating the deubiquitination activity of USP10 and USP13. Cell 2011; 147(1): 223-234.
  35. Jayakiran M. Apoptosis-biochemistry: a mini review. J Clin Exp Pathol 2015; 5(1): 205. doi: 10.4172/2161-0681.1000205.
  36. Khan FH, Dervan E, Bhattacharyya DD, et al. The role of nitric oxide in cancer: master regulator or not? Int J Mol Sci 2020; 21(24): 9393. doi: 10.3390/ijms21249393.
  37. Lis B, Jędrejek D, Stochmal A, et al. Assessment of effects of phenolic fractions from leaves and petals of dandelion in selected components of hemostasis. Food Res Int 2018; 107: 605-612.
  38. Duan X, Pan L, Deng Y, et al. Dandelion root extract affects ESCC progression via regulating multiple signal pathways. Food Funct 2021; 12(19): 9486-9502.
  39. Ovadje P, Roma A, Steckle M, et al. Advances in the research and development of natural health products as main stream cancer therapeutics. Evid Based Complement Altern Med 2015; 2015: 751348. doi: 10.1155/2015/751348.
  40. Trinh NV, Doan-Phuong Dang N, Hong Tran D, et al. Taraxacum officinale dandelion extracts efficiently inhibited the breast cancer stem cell proliferation. Biomed Res Ther 2016; 3(7): 733-741.
Veterinary Research Forum
Volume 14, Issue 9
September 2023
Pages 507-513

  • Receive Date 03 January 2023
  • Revise Date 21 January 2023
  • Accept Date 26 January 2023

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