• Metformin sensitizes gastric cancer cells to chemotherapeutic agents via modulating Shh/Gli1 pathway
  • Mohammad Amin Vatankhah,1 Reza Panahizadeh,2 Mohammad Rahim Vakili,3 Farhad Jeddi,4,* Kazem Nejati-koshki,5 Sina Seifi Mansour,6
    1. Students Research Committee, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran
    2. Students Research Committee, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran
    3. Department of Surgery, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran
    4. Department of Medical Genetics and Pathology, Ardabil University of Medical Sciences, Ardabil, Iran
    5. Pharmaceutical Sciences Research Center, Ardabil University of Medical Sciences, Ardabil, Iran
    6. Students Research Committee, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran


  • Introduction: Gastric cancer (GC) is a crucial cause of cancer-related death characterized by poor prognosis. Docetaxel and 5-fluorouracil (5-FU) are approved for the treatment of GC, but chemo-resistance limits the application of it for GC. Metformin, a popular anti-diabetic drug, has been proven to have potent anticancer effects on gastrointestinal cancers. In this study we investigated the roles of metformin in the chemo-sensitivity of GC cells through targeting Shh/Gli1 Pathway.
  • Methods: Drugs and reagents were purchased from Sigma-Aldrich. The drugs were dissolved in RPMI at the specified concentrations and stored in a 4°C refrigerator. The human AGS cell line purchased from the National Cell Bank of Iran (NCBI, Pasteur Institute, Tehran, Iran). The gastric cancer cells were cultivated in RPMI 1640 (Gibco) medium, which was supplemented with 10% fetal bovine serum (FBS, Gibco) and 100 U/mL penicillin-streptomycin. The cells were incubated in a humidified atmosphere containing 5% CO2. The anticancer effects of metfotmin, 5-FU, docetaxel, and their combination on the AGS gastric cancer cells were evaluated by clonogenic assay and DAPi staining. We used immunocytochemistry assay to assess the expression of the Shh protein. Then, the expression of Gli1, Gli2, and TWIST1 mRNA determined using real-time PCR in these cancerous cells. All data were analyzed using SPSS V.21 software (SPSS Inc., USA). The significant differences were determined using Student’s t-test and one-way ANOVA followed by the Tukey post hoc comparison test (P<0.05).
  • Results: our results demonstrated that metformin increases the sensitivity of GC cells to chemotherapy by enhancing the apoptosis rite and inhibiting clony formation (p<0.05). The co-treatment of GC cells with metformin, 5-FU, and docetaxel attenuated the expression of Shh protein (p<0.05). We also 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 (p<0.05).
  • Conclusion: Overall, our data strongly support an important role for metformin as an enhancer of the efficacy of chemotherapeutic agents against GC via modulating Shh/Gli1 biomarkers.
  • Keywords: gastric cancer, metformin, docetaxel, 5-fluorouracil, cancer biomarcers