Subscribe to RSS
DOI: 10.1055/a-0631-8046
Synergistic Anti-proliferative Effects of Metformin and Silibinin Combination on T47D Breast Cancer Cells via hTERT and Cyclin D1 Inhibition
Publication History
received 31 August 2017
accepted 15 May 2018
Publication Date:
19 June 2018 (online)
![](https://www.thieme-connect.de/media/drugres/201812/lookinside/thumbnails/2017-08-1487_10-1055-a-0631-8046-1.jpg)
Abstract
Background There is a growing body of data that chemotherapeutic combination strategies would be more effective in reducing drug toxicity, inhibiting tumor progression in comparison to either drug alone.
Objective To explore a chemopreventive strategy for improving breast cancer treatment efficacy, the anticancer effects of a combination of Metformin (MET) and Silibinin (SIL) were investigated in T47D breast cancer cells.
Materials and Methods Cytotoxicity of the drugs individually and in combination was evaluated using MTT assay. The precise nature of the interaction between MET and SIL was further analyzed through the median-effect method. In addition, qRT-PCR was applied to determine the expression levels of hTERT and cyclin D1 genes after 48 h drug exposure.
Results MTT assays showed that MET and SIL individually inhibited the cell viability in a dose and time-dependent manner, and the obtained combination indices (CIs) were<1 for all the combination treatments, indicating that the anticancer agents synergistically induced growth inhibition in the breast cancer cells. qPCR findings revealed that the drug combination also synergistically down-regulated the expression levels of hTERT and cyclin D1 at all used concentrations compared with the drugs used alone after 48 h treatment (P≤0.05).
Conclusion The results provide evidence that synergistic antiproliferative effects of MET and SIL, linking to the down-regulation of Cyclin D1 and hTERT genes, and propose that MET+SIL may have therapeutic value in breast cancer therapy.
-
References
- 1 DeSantis CE, Fedewa SA, Goding Sauer A. et al. Breast cancer statistics, 2015: Convergence of incidence rates between black and white women. CA: A Cancer Journal for Clinicians 2016; 66: 31-42
- 2 Runowicz CD, Leach CR, Henry NL. et al. American cancer society/American society of clinical oncology breast cancer survivorship care guideline. CA: A Cancer Journal for Clinicians 2016; 66: 43-73
- 3 Vos S, Van der Groep P, Van der Wall E et al. Hereditary Breast Cancer Syndromes: Molecular Pathogenesis and Diagnostics. eLS 2015
- 4 Maasomi ZJ, Pilehvar-Soltanahmadi Y, Dadashpour M. et al. Synergistic anticancer effects of silibinin and chrysin in T47D breast cancer cells. Asian Pacific Journal of Cancer Prevention: APJCP 2017; 18: 1283
- 5 Farajzadeh R, Pilehvar-Soltanahmadi Y, Dadashpour M. et al. Nano-encapsulated metformin-curcumin in PLGA/PEG inhibits synergistically growth and hTERT gene expression in human breast cancer cells. Artificial Cells, Nanomedicine, and Biotechnology 2017; 1-9
- 6 Alibakhshi A, Ranjbari J, Pilehvar-Soltanahmadi Y. et al. An update on phytochemicals in molecular target therapy of cancer: Potential inhibitory effect on telomerase activity. Current Medicinal Chemistry 2016; 23: 2380-2393
- 7 Mohammadian F, Pilehvar-Soltanahmadi Y, Zarghami F. et al. Upregulation of miR-9 and Let-7a by nanoencapsulated chrysin in gastric cancer cells. Artificial Cells, Nanomedicine, and Biotechnology 2017; 45: 1201-1206
- 8 Montazeri M, Pilehvar-Soltanahmadi Y, Mohaghegh M. et al. Antiproliferative and apoptotic effect of dendrosomal curcumin nanoformulation in P53 mutant and wide-type cancer cell lines. Anti-Cancer Agents in Medicinal Chemistry (Formerly Current Medicinal Chemistry-Anti-Cancer Agents) 2017; 17: 662-673
- 9 Dadashpour M, Pilehvar-Soltanahmadi Y, Zarghami N et al. Emerging importance of phytochemicals in regulation of stem cells fate via signaling pathways. Phytotherapy Research 2017
- 10 Ebrahimnezhad Z, Zarghami N, Keyhani M. et al. Inhibition of hTERT gene expression by silibinin-loaded PLGA-PEG-Fe3O4 in T47D breast cancer cell line. BioImpacts: BI 2013; 3: 67
- 11 Amirsaadat S, Pilehvar-Soltanahmadi Y, Zarghami F. et al. Silibinin-loaded magnetic nanoparticles inhibit hTERT gene expression and proliferation of lung cancer cells. Artificial Cells, Nanomedicine, and Biotechnology 2017; 45: 1649-1656
- 12 Nasiri M, Zarghami N, Koshki KN. et al. Curcumin and silibinin inhibit telomerase expression in T47D human breast cancer cells. Asian Pacific Journal of Cancer Prevention 2013; 14: 3449-3453
- 13 Yurtcu E, Iseri OD, Sahin FI. Effects of silymarin and silymarin-doxorubicin applications on telomerase activity of human hepatocellular carcinoma cell line HepG2. J BUON 2015; 20: 555-561
- 14 Sun H-p, Su J-H, Meng Q-S. et al. Silibinin and indocyanine green-loaded nanoparticles inhibit the growth and metastasis of mammalian breast cancer cells in vitro. Acta Pharmacologica Sinica 2016; 37: 941
- 15 Lotfi-Attari J, Pilehvar-Soltanahmadi Y, Dadashpour M. et al. Co-Delivery of Curcumin and Chrysin by Polymeric Nanoparticles Inhibit Synergistically Growth and hTERT Gene Expression in Human Colorectal Cancer Cells. Nutrition and Cancer 2017; 69: 1290-1299
- 16 Javidfar S, Pilehvar-Soltanahmadi Y, Farajzadeh R. et al. The inhibitory effects of nano-encapsulated metformin on growth and hTERT expression in breast cancer cells. Journal of Drug Delivery Science and Technology 2017
- 17 Pinto AC, Moreira JN, Simões S. Combination chemotherapy in cancer: Principles, evaluation and drug delivery strategies, in Current Cancer Treatment-Novel Beyond Conventional Approaches. 2011, InTech
- 18 Tavakoli F, Jahanban-Esfahlan R, Seidi K. et al. Effects of nano-encapsulated curcumin-chrysin on telomerase, MMPs and TIMPs gene expression in mouse B16F10 melanoma tumour model. Artificial Cells, Nanomedicine, and Biotechnology 2018; 1-12
- 19 Chou T-C. Drug combination studies and their synergy quantification using the Chou-Talalay method. Cancer Research 2010; 70: 440-446
- 20 Burandt E, Grünert M, Lebeau A. et al. Cyclin D1 gene amplification is highly homogeneous in breast cancer. Breast Cancer 2016; 23: 111-119
- 21 Sadeghzadeh H, Pilehvar-Soltanahmad Y, Akbarzadeh A. et al. The effects of nanoencapsulated curcumin-Fe3O4 on proliferation and hTERT gene expression in lung cancer cells. Anti-Cancer Agents in Medicinal Chemistry (Formerly Current Medicinal Chemistry-Anti-Cancer Agents) 2017; 17: 1363-1373
- 22 Pirmoradi S, Fathi E, Farahzadi R. et al. Curcumin affects adipose tissue-derived mesenchymal stem cell aging through TERT gene expression. Drug Research 2018; 68: 213-221
- 23 Kirkpatrick K, Ogunkolade W, Elkak A. et al. hTERT expression in human breast cancer and non-cancerous breast tissue: correlation with tumour stage and c-Myc expression. Breast Cancer Research and Treatment 2003; 77: 277-284
- 24 Mokbel K, Parris CN, Ghilchik M. et al. The association between telomerase, histopathological parameters, and KI-67 expression in breast cancer. The American Journal of Surgery 1999; 178: 69-72
- 25 Wang L, Soria J-C, Kemp BL. et al. hTERT expression is a prognostic factor of survival in patients with stage I non-small cell lung cancer. Clinical Cancer Research 2002; 8: 2883-2889
- 26 Mohammadian F, Abhari A, Dariushnejad H. et al. Upregulation of Mir-34a in AGS gastric cancer cells by a PLGA-PEG-PLGA chrysin nano formulation. Asian Pac J Cancer Prev 2015; 16: 8259-8263
- 27 Fu J, Chou T-C. Abstract 4554 A: Simple, efficient, and quantitative approach for determination of synergism, additive effect, and antagonism of drugs in vivo using combination index method: a proposition for clinical protocol design and regulatory synergy claims. 2017, AACR
- 28 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. Oncology Reports 2004; 11: 493-499
- 29 Iliopoulos D, Hirsch HA, Struhl K. Metformin decreases the dose of chemotherapy for prolonging tumor remission in mouse xenografts involving multiple cancer cell types. Cancer Research 2011; 71: 3196-3201
- 30 Liu H, Scholz C, Zang C. et al. Metformin and the mTOR inhibitor everolimus (RAD001) sensitize breast cancer cells to the cytotoxic effect of chemotherapeutic drugs in vitro. Anticancer Research 2012; 32: 1627-1637
- 31 Soo JS-S, Ng C-H, Tan SH. et al. Metformin synergizes 5-fluorouracil, epirubicin, and cyclophosphamide (FEC) combination therapy through impairing intracellular ATP production and DNA repair in breast cancer stem cells. Apoptosis 2015; 20: 1373-1387
- 32 Vazquez-Martin A, Oliveras-Ferraros C, Del Barco S. et al. et al. The anti-diabetic drug metformin suppresses self-renewal and proliferation of trastuzumab-resistant tumor-initiating breast cancer stem cells. Breast Cancer Research and Treatment 2011; 126: 355-364
- 33 Cufí S, Corominas-Faja B, Vazquez-Martin A. et al. Metformin-induced preferential killing of breast cancer initiating CD44+ CD24−/low cells is sufficient to overcome primary resistance to trastuzumab in HER2+ human breast cancer xenografts. Oncotarget 2012; 3: 395
- 34 Ma J, Guo Y, Chen S. et al. Metformin enhances tamoxifen-mediated tumor growth inhibition in ER-positive breast carcinoma. BMC Cancer 2014; 14: 172
- 35 Jang SY, Kim A, Kim JK. et al. Metformin inhibits tumor cell migration via down-regulation of MMP9 in tamoxifen-resistant breast cancer cells. Anticancer Research 2014; 34: 4127-4134
- 36 Rocha GZ, Dias MM, Ropelle ER. et al. Metformin amplifies chemotherapy-induced AMPK activation and antitumoral growth. Clinical Cancer Research 2011; 17: 3993-4005
- 37 Cantrell LA, Zhou C, Mendivil A. et al. Metformin is a potent inhibitor of endometrial cancer cell proliferation—implications for a novel treatment strategy. Gynecologic Oncology 2010; 116: 92-98
- 38 Kim S, Choi JH, Lim HI. et al. Silibinin prevents TPA-induced MMP-9 expression and VEGF secretion by inactivation of the Raf/MEK/ERK pathway in MCF-7 human breast cancer cells. Phytomedicine 2009; 16: 573-580
- 39 Lu W, Lin C, King TD. et al. Silibinin inhibits Wnt/β-catenin signaling by suppressing Wnt co-receptor LRP6 expression in human prostate and breast cancer cells. Cellular Signalling 2012; 24: 2291-2296