Download PDF
Drug Res (Stuttg) 2021; 71(01): 17-25
DOI: 10.1055/a-1248-9008
Original Article

Efficacy of Metformin and Chemotherapeutic Agents on the Inhibition of Colony Formation and Shh/Gli1 Pathway: Metformin/Docetaxel Versus Metformin/5-Fluorouracil

Maryam Fatehi-Agdam
1   Students Research Committee, School of Medicine, Ardabil University of Medical Sciences, Ardabil
2   Research Laboratory for Embryology and Stem Cells, Department of Anatomical Sciences, School of Medicine, Ardabil University of Medical Sciences, Ardabil
,
Mohammad Amin Vatankhah
1   Students Research Committee, School of Medicine, Ardabil University of Medical Sciences, Ardabil
2   Research Laboratory for Embryology and Stem Cells, Department of Anatomical Sciences, School of Medicine, Ardabil University of Medical Sciences, Ardabil
,
Reza Panahizadeh 
1   Students Research Committee, School of Medicine, Ardabil University of Medical Sciences, Ardabil
2   Research Laboratory for Embryology and Stem Cells, Department of Anatomical Sciences, School of Medicine, Ardabil University of Medical Sciences, Ardabil
,
Farhad Jeddi
2   Research Laboratory for Embryology and Stem Cells, Department of Anatomical Sciences, School of Medicine, Ardabil University of Medical Sciences, Ardabil
,
Nowruz Najafzadeh 
2   Research Laboratory for Embryology and Stem Cells, Department of Anatomical Sciences, School of Medicine, Ardabil University of Medical Sciences, Ardabil
› Author Affiliations Funding: This work was supported by a thesis grant for M.Sc. (IR.ARUMS.REC.1397.240) and a medical student research grant (IR.ARUMS.REC.1398.418) from Ardabil University of Medical Sciences.
› Further Information
Also available at
   Permissions and Reprints

Abstract

Background Gastric cancer is a common gastrointestinal cancer characterized by poor prognosis and chemoresistance. Docetaxel and 5-fluorouracil (5-FU) are frequently used for the treatment of gastric cancer. Despite their potent anti-cancer effects, chemoresistance occurs in metastatic gastric cancer. Metformin, a popular anti-diabetic drug, has been proven to have potent anticancer effects on gastrointestinal cancers. Here, we aim to improve this chemotherapy agents’ efficacy by pretreatment with metformin.

Methods The AGS gastric cancer cell line were pretreated with three different sub-toxic concentration of metformin and then treated with various concentrations of 5-FU and docetaxel.

The anticancer effects of the combination of metformin with the chemotherapy agents were determined using clonogenic assay and DAPi staining. We used real-time PCR to evaluate Gli1, Gli2, and TWIST1 mRNA expression levels in the gastric cancer cells. Also, the expression of the Shh protein was assessed using immunocytochemistry.

Results Here, we found that metformin sensitized the gastric cancer cells to chemotherapy. The combination treatments were more effective in reducing the number of cancer colonies compared to 5-FU or docetaxel alone. The combination of metformin with 5-FU or docetaxel significantly reduced the number of cells expressing the Shh protein compared to the 5-FU alone or docetaxel alone. Interestingly, we found that the combination of metformin with docetaxel significantly down-regulated the mRNA levels of Gli1, Gli2, and TWIST1 in the AGS gastric cancer cell line compared to docetaxel alone.

Conclusion Overall, our data strongly support an important role for metformin as an enhancer of the efficacy of chemotherapeutic agents against gastric cancer.

Key words

metformin - docetaxel - 5-fluorouracil - Shh - Gli1 - Gli2 - gastric cancer


Publication History

Received: 22 July 2020

Accepted: 24 August 2020

Article published online:
28 September 2020

© 2020. Thieme. All rights reserved.

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

 

  • References

  • 1 Etemadi A, Safiri S, Sepanlou SG. et al. The global, regional, and national burden of stomach cancer in 195 countries, 1990–2017: A systematic analysis for the Global Burden of Disease study 2017. Lancet Gastroenterol Hepatol 2020; 5: 42-54
    CrossrefPubMedGoogle Scholar
  • 2 Rawla P, Barsouk A. Epidemiology of gastric cancer: Global trends, risk factors and prevention. Prz Gastroenterol 2019; 14: 26
    PubMedGoogle Scholar
  • 3 Bilici A. Treatment options in patients with metastatic gastric cancer: Current status and future perspectives. World J Gastroenterol 2014; 20: 3905
    CrossrefPubMedGoogle Scholar
  • 4 Buckland G, Travier N, Huerta J. et al. Healthy lifestyle index and risk of gastric adenocarcinoma in the EPIC cohort study. Int J Cancer 2015; 137: 598-606
    CrossrefPubMedGoogle Scholar
  • 5 de Weger VA, Beijnen JH, Schellens JH. Cellular and clinical pharmacology of the taxanes docetaxel and paclitaxel–a review. Anti-cancer drugs 2014; 25: 488-494
    CrossrefPubMedGoogle Scholar
  • 6 Kim R, Ohi Y, Inoue H. et al. Taxotere activates transcription factor AP-1 in association with apoptotic cell death in gastric cancer cell lines. Anticancer res 1999; 19: 5399-5405
    PubMedGoogle Scholar
  • 7 Kim NK, Park YS, Heo DS. et al. A phase III randomized study of 5-fluorouracil and cisplatin versus 5-fluorouracil, doxorubicin, and mitomycin C versus 5-fluorouracil alone in the treatment of advanced gastric cancer. Cancer 1993; 71: 3813-3818
    CrossrefPubMedGoogle Scholar
  • 8 Rena G, Hardie DG, Pearson ER. The mechanisms of action of metformin. Diabetologia 2017; 60: 1577-1585
    CrossrefPubMedGoogle Scholar
  • 9 Kasznicki J, Sliwinska A, Drzewoski J. Metformin in cancer prevention and therapy. Ann Trans Med 2014; 2: 57
    PubMedGoogle Scholar
  • 10 Honjo S, Ajani JA, Scott AW. et al. Metformin sensitizes chemotherapy by targeting cancer stem cells and the mTOR pathway in esophageal cancer. Int J Oncol 2014; 45: 567-574
    CrossrefPubMedGoogle Scholar
  • 11 Liu X, Yun F, Shi L. et al. Roles of signaling pathways in the epithelial-mesenchymal transition in cancer. Asian Pac J Cancer Prev 2015; 16: 6201-6206
    CrossrefPubMedGoogle Scholar
  • 12 Huang L, Wu R-L, Xu A-M. Epithelial-mesenchymal transition in gastric cancer. Am J Transl Res 2015; 7: 2141
    PubMedGoogle Scholar
  • 13 Hsu K-W, Hsieh R-H, Huang K-H. et al. Activation of the Notch1/STAT3/Twist signaling axis promotes gastric cancer progression. Carcinogenesis 2012; 33: 1459-1467
    CrossrefPubMedGoogle Scholar
  • 14 Fatehi Aghdam M, Najafzadeh NMGG. Evaluation of cytotoxic effects of the combination of metformin with docetaxel and 5-fluorouracil on the gastric cancer cells. Journal of Isfahan Medical School 2020; 38: 139-146
    PubMedGoogle Scholar
  • 15 Mohammadi Jobani B, Najafzadeh N, Mazani M. et al. Molecular mechanism and cytotoxicity of allicin and all-trans retinoic acid against CD44+ versus CD117+ melanoma cells. Phytomedicine 2018; 48: 161-169
    CrossrefPubMedGoogle Scholar
  • 16 Rosenberg AJ, Rademaker A, Hochster HS. et al. Docetaxel, Oxaliplatin, and 5-Fluorouracil (DOF) in Metastatic and Unresectable Gastric/Gastroesophageal Junction Adenocarcinoma: A Phase II Study with Long-Term Follow-Up. The oncologist 2019; 24: 1039
    CrossrefPubMedGoogle Scholar
  • 17 Fan C, Wang Y, Liu Z. et al. Metformin exerts anticancer effects through the inhibition of the Sonic hedgehog signaling pathway in breast cancer. Int J Mol Med 2015; 36: 204-214
    CrossrefPubMedGoogle Scholar
  • 18 Islam S, Mokhtari R, Noman A. et al. Sonic hedgehog (Shh) signaling promotes tumorigenicity and stemness via activation of epithelial-to-mesenchymal transition (EMT) in bladder cancer. Mol Carcinog 2016; 55: 537-551
    CrossrefPubMedGoogle Scholar
  • 19 Lees C, Howie S, Sartor RB. et al. The hedgehog signalling pathway in the gastrointestinal tract: Implications for development, homeostasis, and disease. Gastroenterology 2005; 129: 1696-1710
    CrossrefPubMedGoogle Scholar
  • 20 Sanchez P, Hernández AM, Stecca B. et al. Inhibition of prostate cancer proliferation by interference with SONIC HEDGEHOG-GLI1 signaling. Proc Natl Acad Sci USA 2004; 101: 12561-12566
    CrossrefPubMedGoogle Scholar
  • 21 Shahi MH, Rey JA, Castresana JS. The sonic hedgehog-GLI1 signaling pathway in brain tumor development. Expert Opin Ther Targets 2012; 16: 1227-1238
    CrossrefPubMedGoogle Scholar
  • 22 Song Z, Wei B, Lu C. et al. Metformin suppresses the expression of Sonic hedgehog in gastric cancer cells. Mol Med Rep 2017; 15: 1909-1915
    CrossrefPubMedGoogle Scholar
  • 23 Gonnissen A, Isebaert S, McKee CM. et al. The effect of metformin and GANT61 combinations on the radiosensitivity of prostate cancer cells. Int J Mol Sci 2017; 18: 399
    CrossrefPubMedGoogle Scholar
  • 24 Yu Y, Fang W, Xia T. et al. Metformin potentiates rapamycin and cisplatin in gastric cancer in mice. Oncotarget 2015; 6: 12748
    CrossrefPubMedGoogle Scholar
  • 25 Wandee J, Prawan A, Senggunprai L. et al. Metformin sensitizes cholangiocarcinoma cell to cisplatin-induced cytotoxicity through oxidative stress mediated mitochondrial pathway. Life Sci 2019; 217: 155-163
    CrossrefPubMedGoogle Scholar
  • 26 dos Santos Guimarães I, Ladislau-Magescky T, Tessarollo NG. et al. Chemosensitizing effects of metformin on cisplatin-and paclitaxel-resistant ovarian cancer cell lines. Pharmacol Rep 2018; 70: 409-417
    CrossrefPubMedGoogle Scholar
  • 27 Peng M, Darko KO, Tao T. et al. Combination of metformin with chemotherapeutic drugs via different molecular mechanisms. Cancer Treat Rev 2017; 54: 24-33
    CrossrefPubMedGoogle Scholar
  • 28 Zhang Y, Guan M, Zheng Z. et al. Effects of metformin on CD133+ colorectal cancer cells in diabetic patients. PLoS One 2013; 8 (11) e81264
    CrossrefPubMedGoogle Scholar
  • 29 Yang X, Sun D, Tian Y. et al. Metformin sensitizes hepatocellular carcinoma to arsenic trioxide-induced apoptosis by downregulating Bcl2 expression. Tumor Biol 2015; 36: 2957-2964
    CrossrefPubMedGoogle Scholar
  • 30 Zakikhani M, Blouin M-J, Piura E. et al. Metformin and rapamycin have distinct effects on the AKT pathway and proliferation in breast cancer cells. Breast Cancer Res Treat 2010; 123: 271-279
    CrossrefPubMedGoogle Scholar
  • 31 Mayer MJ, Klotz LH, Venkateswaran V. The effect of metformin use during docetaxel chemotherapy on prostate cancer specific and overall survival of diabetic patients with castration resistant prostate cancer. J Urol 2017; 197: 1068-1075
    CrossrefPubMedGoogle Scholar
  • 32 Lesan V, Ghaffari SH, Salaramoli J. et al. Evaluation of antagonistic effects of metformin with Cisplatin in gastric cancer cells. Int J Hematol Oncol Stem Cell Res 2014; 8: 12
    PubMedGoogle Scholar
  • 33 Zhou C, Li X, Shan S. et al. Metformin’s effect on 5-fluorouracil, cisplatin, paclitaxel in laryngocarcinoma Hep-2 cells. J Otorhinolaryngol Head Neck Surg 2017; 31: 524-528
    PubMedGoogle Scholar
  • 34 Lu J-t, He W, Wei W. Hedgehog signaling pathway mediates invasion and metastasis of hepatocellular carcinoma via ERK pathway. Acta Pharmacol Sin 2012; 33: 691-700
    CrossrefPubMedGoogle Scholar
  • 35 Ma X, Chen K, Huang S. et al. Frequent activation of the hedgehog pathway in advanced gastric adenocarcinomas. Carcinogenesis 2005; 26: 1698-1705
    CrossrefPubMedGoogle Scholar
  • 36 Yao Y, Zhou D, Shi D. et al. GLI1 overexpression promotes gastric cancer cell proliferation and migration and induces drug resistance by combining with the AKT-mTOR pathway. Biomed Pharmacother 2019; 111: 993-1004
    CrossrefPubMedGoogle Scholar
  • 37 Yu D, Shin H-S, Lee YS. et al. Genistein attenuates cancer stem cell characteristics in gastric cancer through the downregulation of Gli1. Oncol Rep 2014; 31: 673-678
    CrossrefPubMedGoogle Scholar