Administration of Gapmer-type Antisense Oligonucleotides Targeting γ-Glutamylcyclotransferase Suppresses the Growth of A549 Lung Cancer Xenografts

Anticancer Res. 2022 Mar;42(3):1221-1227. doi: 10.21873/anticanres.15589.

Abstract

Background/aim: γ-Glutamyl cyclotransferase (GGCT) is up-regulated in various cancer types, including lung cancer. In this study, we evaluated efficacy of gapmer-type antisense oligonucleotides (ASOs) targeting GGCT in an A549 lung cancer xenograft mouse model and studied their mechanisms of action.

Materials and methods: GGCT was inhibited using GGCT-ASOs and cell proliferation was evaluated by dye exclusion test. Western blot analysis was conducted to measure expression of GGCT, p21, p16 and p27, phosphorylation of AMP-activated protein kinase, and caspase activation in A549 cells. Induction of apoptosis and up-regulation of reactive oxygen species were assessed by flow cytometry using annexin V staining and 2',7'-dichlorodihydrofluorescein diacetate dye, respectively.

Results: GGCT-ASOs suppressed GGCT expression in A549 cells, inhibited proliferation, and induced apoptosis with activation of caspases. GGCT-ASOs also increased expression of cell-cycle regulating proteins, phospho-AMPK and ROS levels. Systemic administration of GGCT-ASOs to animals bearing A549 lung cancer xenografts showed significant antitumor effects without evident toxicity.

Conclusion: GGCT-ASOs appear to be promising as novel cancer therapeutic agents.

Keywords: AMPK; antisense; cyclin-dependent kinase inhibitor; lung cancer; reactive oxygen species; γ-glutamylcyclotransferase.

MeSH terms

  • A549 Cells
  • Animals
  • Antineoplastic Agents / pharmacology*
  • Apoptosis
  • Caspases / metabolism
  • Cell Cycle Proteins / metabolism
  • Cell Proliferation / drug effects*
  • Cycloheximide / analogs & derivatives
  • Cycloheximide / metabolism
  • Gene Expression Regulation, Neoplastic
  • Humans
  • Lung Neoplasms / drug therapy*
  • Lung Neoplasms / enzymology
  • Lung Neoplasms / genetics
  • Lung Neoplasms / pathology
  • Male
  • Mice
  • Mice, SCID
  • Oligonucleotides, Antisense / pharmacology*
  • Signal Transduction
  • Tumor Burden
  • Xenograft Model Antitumor Assays
  • gamma-Glutamylcyclotransferase / genetics
  • gamma-Glutamylcyclotransferase / metabolism*

Substances

  • Antineoplastic Agents
  • Cell Cycle Proteins
  • GGCT protein, human
  • Oligonucleotides, Antisense
  • streptovitacin A
  • Cycloheximide
  • Caspases
  • gamma-Glutamylcyclotransferase