Autophagy Facilitates Metadherin-Induced Chemotherapy Resistance Through the AMPK/ATG5 Pathway in Gastric Cancer

Cell Physiol Biochem. 2018;46(2):847-859. doi: 10.1159/000488742. Epub 2018 Apr 9.

Abstract

Background/aims: Metadherin (MTDH) is overexpressed in some malignancies and enhances drug resistance; however, its role in gastric cancer (GC) and the underlying mechanisms remain largely unexplored. Here, we explore the mechanism by which MTDH induces drug resistance in GC.

Methods: We analysed the level of MTDH in GC and adjacent normal gastric mucosal tissues by real-time quantitative PCR (q-PCR). We also analysed the level of autophagy by western blot analysis, confocal microscopy, and transmission electron microscopy after MTDH knockdown and overexpression, and examined fluorouracil (5-FU) resistance by Cell Counting Kit-8 at the same time. Finally, GC patient-derived xenograft tumours were used to demonstrate 5-FU resistance. An AMPK pathway inhibitor was applied to determine the molecular mechanisms of autophagy.

Results: MTDH expression was significantly increased in the GC specimens compared with that in the adjacent normal gastric mucosal tissues. Further study showed a positive correlation between the expression level of MTDH and 5-FU resistance. MTDH overexpression in MKN45 cells increased the levels of P-glycoprotein (P-gp) and promoted 5-FU resistance, while inhibition of MTDH showed the opposite result. The simultaneous inhibition of autophagy and overexpression of MTDH decreased the levels of P-gp and inhibited 5-FU resistance. Moreover, MTDH induced AMPK phosphorylation, regulated ATG5 expression, and finally influenced autophagy, suggesting that MTDH may activate autophagy via the AMPK/ATG5 signalling pathway. Our findings reveal a unique mechanism by which MTDH promotes GC chemoresistance and show that MTDH is a potential target for improved chemotherapeutic sensitivity and GC patient survival.

Conclusions: MTDH-stimulated cancer resistance to 5-FU may be mediated through autophagy activated by the AMPK/ATG5 pathway in GC.

Keywords: Autophagy; Chemoresistance; Gastric cancer; Mtdh.

MeSH terms

  • AMP-Activated Protein Kinases / antagonists & inhibitors
  • AMP-Activated Protein Kinases / genetics
  • AMP-Activated Protein Kinases / metabolism*
  • ATP Binding Cassette Transporter, Subfamily B, Member 1 / metabolism
  • Animals
  • Autophagy* / drug effects
  • Autophagy-Related Protein 5 / metabolism*
  • Cell Adhesion Molecules / antagonists & inhibitors
  • Cell Adhesion Molecules / genetics
  • Cell Adhesion Molecules / metabolism*
  • Cell Line, Tumor
  • Drug Resistance, Neoplasm
  • Fluorouracil / therapeutic use
  • Fluorouracil / toxicity
  • Humans
  • Immunohistochemistry
  • Male
  • Membrane Proteins
  • Mice
  • Mice, Inbred BALB C
  • Microtubule-Associated Proteins / metabolism
  • Phosphorylation
  • RNA Interference
  • RNA, Small Interfering / metabolism
  • RNA-Binding Proteins
  • Signal Transduction
  • Stomach Neoplasms / drug therapy
  • Stomach Neoplasms / metabolism
  • Stomach Neoplasms / pathology*
  • Transplantation, Heterologous

Substances

  • ATP Binding Cassette Transporter, Subfamily B, Member 1
  • Autophagy-Related Protein 5
  • Cell Adhesion Molecules
  • MAP1LC3A protein, human
  • MTDH protein, human
  • Membrane Proteins
  • Microtubule-Associated Proteins
  • RNA, Small Interfering
  • RNA-Binding Proteins
  • AMP-Activated Protein Kinases
  • Fluorouracil