ECRG2 enhances the anti-cancer effects of cisplatin in cisplatin-resistant esophageal cancer cells via upregulation of p53 and downregulation of PCNA

World J Gastroenterol. 2017 Mar 14;23(10):1796-1803. doi: 10.3748/wjg.v23.i10.1796.

Abstract

Aim: To explore the anti-tumor effects of esophageal cancer-related gene 2 (ECRG2) in combination with cisplatin (DDP) in DDP-resistant esophageal cancer cells (EC9706/DDP).

Methods: A drug-resistant cell model was established, with EC9706/DDP cells being treated with ECRG2 and/or DDP. Cell viability was examined by MTT assay. The rate of cell apoptosis was determined by flow cytometry. The mRNA expression levels of proliferating cell nuclear antigen (PCNA), metallothionein (MT), and p53 were determined by RT-PCR and PCNA, while MT and p53 protein expression levels were determined by western blotting.

Results: The anti-proliferative effect of ECRG2 in combination with DDP was superior when compared to ECRG2 or DDP alone. The inhibition rate for the combination reached its peak (51.33%) at 96 h. The early apoptotic rates of the control, ECRG2 alone, DDP alone, and ECRG2 plus DDP groups were 5.71% ± 0.27%, 12.68% ± 0.61%, 14.15% ± 0.87%, and 27.96% ± 0.36%, respectively. Although all treatment groups were significantly different from the control group (P < 0.05), the combination treatment of ECRG2 plus DDP performed significantly better when compared to either ECRG2 or DDP alone (P < 0.05). The combination of ECRG2 and DDP significantly upregulated p53 mRNA and protein levels and downregulated PCNA mRNA and protein levels compared to ECRG2 or DDP alone (P < 0.05). However, no changes were seen in the expression of MT mRNA or protein.

Conclusion: ECRG2 in combination with DDP can inhibit viability and induce apoptosis in esophageal cancer DDP-resistant cells, possibly via upregulation of p53 expression and downregulation of PCNA expression. These findings suggest that the combination of ECRG2 and DDP may be a promising strategy for the clinical treatment of esophageal cancers that are resistant to DDP.

Keywords: Cisplatin; Esophageal cancer related-gene 2; Proliferating cell nuclear antigen; Resistance; p53.

MeSH terms

  • Antineoplastic Agents / pharmacology*
  • Antineoplastic Agents / therapeutic use
  • Apoptosis / drug effects
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Cisplatin / pharmacology*
  • Cisplatin / therapeutic use
  • Down-Regulation
  • Drug Resistance, Neoplasm / drug effects*
  • Drug Therapy, Combination
  • Esophageal Neoplasms / drug therapy*
  • Esophageal Neoplasms / physiopathology
  • Flow Cytometry
  • Humans
  • Metallothionein / metabolism
  • Proliferating Cell Nuclear Antigen / metabolism*
  • Proteinase Inhibitory Proteins, Secretory / pharmacology
  • Proteinase Inhibitory Proteins, Secretory / therapeutic use*
  • RNA, Messenger / metabolism
  • Recombinant Proteins / pharmacology
  • Recombinant Proteins / therapeutic use
  • Serine Peptidase Inhibitors, Kazal Type
  • Transcriptional Activation
  • Tumor Suppressor Protein p53 / metabolism*
  • Up-Regulation

Substances

  • Antineoplastic Agents
  • Proliferating Cell Nuclear Antigen
  • Proteinase Inhibitory Proteins, Secretory
  • RNA, Messenger
  • Recombinant Proteins
  • SPINK7 protein, human
  • Serine Peptidase Inhibitors, Kazal Type
  • TP53 protein, human
  • Tumor Suppressor Protein p53
  • Metallothionein
  • Cisplatin