Silibinin, Synergistically Enhances Vinblastine-Mediated Apoptosis in Triple Negative Breast Cancer Cell Line: Involvement of Bcl2/Bax and Caspase-3 Pathwa
-
Hassan Dariushnejad
,
Neda Roshanravan
,
Hunar Mustafa Wasman
,
Mostafa Cheraghi
,
Lale Pirzeh
,
Vajihe Ghorbanzadeh
Abstract
Background: Triple-negative breast cancer (TNBC) with a poor prognosis and survival is the most invasive subtype of breast cancer. Usually, TNBC requires a chemotherapy regimen at all stages, but chemotherapy drugs have shown many side effects. We assumed that combination therapy of vinblastine and silibinin might reduce the vinblastine toxicity and dose of vinblastine.
Materials and Methods: The MDA-MB-231 were cells subjected to MTT assay for IC50 determination and combination effects, which were measured based on Chou-Talalay's method. The type of cell death was determined by using a Flow-cytometric assay. Cell death pathway markers, including Bcl-2, Bax, and caspase-3 were analyzed by western blot and Real-Time PCR.
Results: The treatment of MDA-MB-231 cells exhibited IC50 and synergism at the combination of 30 µM of silibinin and 4 µm of vinblastine in cell viability assay (CI=0.69). YO-PRO-1/PI double staining results showed a significant induction of apoptosis when MDA-MB-231 cells were treated with a silibinin and vinblastine combination (p<0.01). Protein levels of Bax and cleaved caspase-3 were significantly upregulated, and Bcl-2 downregulated significantly. Significant upregulation of Bax (2.96-fold) and caspase-3 (3.46-fold) while Bcl-2 was downregulated by 2-fold.
Conclusion: Findings established a preclinical rationale for the combination of silibinin and vinblastine. This combination produces synergistic effects in MDA-MB-231 cells by altering pro- and anti-apoptotic genes, which may reduce the toxicity and side effects of vinblastine.
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13. Chavoshi H, Vahedian V, Saghaei S, et al. Adjuvant therapy with silibinin improves the efficacy of paclitaxel and cisplatin in MCF-7 breast cancer cells. Asian Pac J Cancer Prev. 2017;18(8):2243-2247.
14. Chou TC, Talalay P. Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul.1984:22:27-55.
15. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods. 2001; 25(4):402-8.
16. Brandl MB, Pasquier E, Li F, et al. Computational analysis of image-based drug profiling predicts synergistic drug combinations: Applications in triple-negative breast cancer. Mol Oncol. 2014;8(8):1548-60.
17. Li J, Chen S, Chen C, et al. Pathological complete response as a surrogate for relapse-free survival in patients with triple negative breast cancer after neoadjuvant chemotherapy. Oncotarget. 2017; 8(11):18399-18408.
18. Choudhari AS, Mandave PC, Deshpande M, et al. Phytochemicals in cancer treatment: From preclinical studies to clinical practice. Front Pharmacol. 2020:10:1614.
19. Gigant B, Wang C, Ravelli RB, et al. Structural basis for the regulation of tubulin by vinblastine. Nature. 2005; 435(7041):519-22.
20. Pfister C, Gravis G, Fléchon A, et al. Randomized phase III trial of dose-dense Methotrexate, Vinblastine, Doxorubicin, and Cisplatin, or Gemcitabine and Cisplatin as perioperative chemotherapy for patients with muscle-invasive bladder cancer. Analysis of the GETUG/AFU V05 VESPER trial secondary endpoints: Chemotherapy toxicity and pathological responses. Eur Urol. 2021;79(2):214-221.
21. Nejati-Koshki K, Akbarzadeh A, Pourhasan-Moghaddam M, et al. Inhibition of leptin and leptin receptor gene expression by silibinin-curcumin combination. Asian Pac J Cancer Prev. 2014;14(11):6595-9.
22. Esmaeil Lashgarian H, Adamii V, Ghorbanzadeh V, et al. Silibinin Inhibit Cell Migration through Downregulation of RAC1 Gene Expression in Highly Metastatic Breast Cancer Cell Line. Drug Res (Stuttg). 2020;70(10):478-483.
23. Tuli HS, Mittal S, Aggarwal D, et al. Path of Silibinin from diet to medicine: A dietary polyphenolic flavonoid having potential anti-cancer therapeutic significance. Semin Cancer Biol. 2021:73:196-218.
24. Tyagi AK, Agarwal C, Chan DC, et al. Synergistic anti-cancer effects of silibinin with conventional cytotoxic agents doxorubicin, cisplatin and carboplatin against human breast carcinoma MCF-7 and MDA-MB468 cells. Oncol Rep. 2004;11(2):493-9.
25. Xu G, Shi Y. Apoptosis signaling pathways and lymphocyte homeostasis. Cell Res. 2007;17(9):759-71.
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27. Lee JW, Park S, Kim SY, et al. Curcumin hampers the antitumor effect of vinblastine via the inhibition of microtubule dynamics and mitochondrial membrane potential in HeLa cervical cancer cells. Phytomedicine. 2016; 23(7):705-13.
28. Li L, Zeng J, Gao Y, et al. Targeting silibinin in the antiproliferative pathway. Expert Opin Investig Drugs. 2010;19(2):243-55.
29. Wang X, Wang D, Zhao Y. Effect and mechanism of resveratrol on the apoptosis of lung adenocarcinoma cell line A549. Cell Biochem Biophys. 2015;73(2):527-531.
30. Fan S, Qi M, Yu Y, et al. P53 activation plays a crucial role in silibinin induced ROS generation via PUMA and JNK. Free Radic Res. 2012;46(3):310-9.
31. D’Arcy MS. Cell death: a review of the major forms of apoptosis, necrosis and autophagy. Cell Biol Int. 2019;43(6):582-592.
2. Zhou Y, Yamamoto Y, Takeshita F, et al. Delivery of miR-424-5p via Extracellular vesicles promotes the apoptosis of MDA-MB-231 TNBC cells in the tumor microenvironment. Int J Mol Sci. 2021;22(2):844.
3. Bai X, Ni J, Beretov J, et al. Triple-negative breast cancer therapeutic resistance: Where is the Achilles' heel? Cancer Lett. 2021; 497:100-11.
4. El‐Sayed ESR. Discovery of the Anticancer Drug Vinblastine from the Endophytic Alternaria alternata and Yield Improvement by Gamma Irradiation Mutagenesis. J Appl Microbiol. 2021;131(6):2886-2898.
5. Aggarwal S, Verma SS, Aggarwal S, et al. Drug repurposing for breast cancer therapy: Old weapon for new battle. Semin Cancer Biol. 2021:68:8-20.
6. Aramjoo H, Zadeh HE, Hemmati M. Investigation of Protective Effects of Quercetin on Oxidative Stress Induced by Vinblastine in Bone Marrow of Rats. Pharm Sci. 2021;27(2):194-200.
7. Zhou H, Liu L, Ma X, et al. RIP1/RIP3/MLKL-mediated necroptosis contributes to vinblastine-induced myocardial damage. Mol Cell Biochem. 2021; 476(2): 1233–1243.
8. Baksi R, Rana R, Nivsarkar M. Chemopreventive potential of plant-derived epigenetic inhibitors silibinin and quercetin: an involvement of apoptotic signaling cascade modulation. Future J Pharm Sci. 2021; 7(69):1-13.
9. Delmas D, Xiao J, Vejux A, et al. Silymarin and cancer: A dual strategy in both in chemoprevention and chemosensitivity. Molecules. 2020; 25(9):2009.
10. Mao J, Yang H, Cui T, et al. Combined treatment with sorafenib and silibinin synergistically targets both HCC cells and cancer stem cells by enhanced inhibition of the phosphorylation of STAT3/ERK/AKT. Eur J Pharmacol. 2018:832:39-49.
11. Tiwari P, Mishra K. Silibinin in cancer therapy: A promising prospect. Cancer Res Front. 2015; 1(3):303-18.
12. Sak K. Chemotherapy and dietary phytochemical agents. Chemotherapy research and practice. Chemother Res Pract. 2012:2012:282570.
13. Chavoshi H, Vahedian V, Saghaei S, et al. Adjuvant therapy with silibinin improves the efficacy of paclitaxel and cisplatin in MCF-7 breast cancer cells. Asian Pac J Cancer Prev. 2017;18(8):2243-2247.
14. Chou TC, Talalay P. Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul.1984:22:27-55.
15. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods. 2001; 25(4):402-8.
16. Brandl MB, Pasquier E, Li F, et al. Computational analysis of image-based drug profiling predicts synergistic drug combinations: Applications in triple-negative breast cancer. Mol Oncol. 2014;8(8):1548-60.
17. Li J, Chen S, Chen C, et al. Pathological complete response as a surrogate for relapse-free survival in patients with triple negative breast cancer after neoadjuvant chemotherapy. Oncotarget. 2017; 8(11):18399-18408.
18. Choudhari AS, Mandave PC, Deshpande M, et al. Phytochemicals in cancer treatment: From preclinical studies to clinical practice. Front Pharmacol. 2020:10:1614.
19. Gigant B, Wang C, Ravelli RB, et al. Structural basis for the regulation of tubulin by vinblastine. Nature. 2005; 435(7041):519-22.
20. Pfister C, Gravis G, Fléchon A, et al. Randomized phase III trial of dose-dense Methotrexate, Vinblastine, Doxorubicin, and Cisplatin, or Gemcitabine and Cisplatin as perioperative chemotherapy for patients with muscle-invasive bladder cancer. Analysis of the GETUG/AFU V05 VESPER trial secondary endpoints: Chemotherapy toxicity and pathological responses. Eur Urol. 2021;79(2):214-221.
21. Nejati-Koshki K, Akbarzadeh A, Pourhasan-Moghaddam M, et al. Inhibition of leptin and leptin receptor gene expression by silibinin-curcumin combination. Asian Pac J Cancer Prev. 2014;14(11):6595-9.
22. Esmaeil Lashgarian H, Adamii V, Ghorbanzadeh V, et al. Silibinin Inhibit Cell Migration through Downregulation of RAC1 Gene Expression in Highly Metastatic Breast Cancer Cell Line. Drug Res (Stuttg). 2020;70(10):478-483.
23. Tuli HS, Mittal S, Aggarwal D, et al. Path of Silibinin from diet to medicine: A dietary polyphenolic flavonoid having potential anti-cancer therapeutic significance. Semin Cancer Biol. 2021:73:196-218.
24. Tyagi AK, Agarwal C, Chan DC, et al. Synergistic anti-cancer effects of silibinin with conventional cytotoxic agents doxorubicin, cisplatin and carboplatin against human breast carcinoma MCF-7 and MDA-MB468 cells. Oncol Rep. 2004;11(2):493-9.
25. Xu G, Shi Y. Apoptosis signaling pathways and lymphocyte homeostasis. Cell Res. 2007;17(9):759-71.
26. Sharma A, Kaur M, Katnoria JK, et al. Polyphenols in food: Cancer prevention and apoptosis induction. Curr Med Chem. 2018;25(36):4740-4757.
27. Lee JW, Park S, Kim SY, et al. Curcumin hampers the antitumor effect of vinblastine via the inhibition of microtubule dynamics and mitochondrial membrane potential in HeLa cervical cancer cells. Phytomedicine. 2016; 23(7):705-13.
28. Li L, Zeng J, Gao Y, et al. Targeting silibinin in the antiproliferative pathway. Expert Opin Investig Drugs. 2010;19(2):243-55.
29. Wang X, Wang D, Zhao Y. Effect and mechanism of resveratrol on the apoptosis of lung adenocarcinoma cell line A549. Cell Biochem Biophys. 2015;73(2):527-531.
30. Fan S, Qi M, Yu Y, et al. P53 activation plays a crucial role in silibinin induced ROS generation via PUMA and JNK. Free Radic Res. 2012;46(3):310-9.
31. D’Arcy MS. Cell death: a review of the major forms of apoptosis, necrosis and autophagy. Cell Biol Int. 2019;43(6):582-592.
| Files | ||
| Issue | Vol 18 No 2 (2024) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijhoscr.v18i2.15375 | |
| Keywords | ||
| Vinblastine Silibini Drug combination Triple negative breast cancer (TNBC) Apoptosis | ||
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How to Cite
1.
Dariushnejad H, Roshanravan N, Wasman HM, Cheraghi M, Pirzeh L, Ghorbanzadeh V. Silibinin, Synergistically Enhances Vinblastine-Mediated Apoptosis in Triple Negative Breast Cancer Cell Line: Involvement of Bcl2/Bax and Caspase-3 Pathwa. Int J Hematol Oncol Stem Cell Res. 2024;18(2):174-182.
