Genetically engineered drug rhCNB induces apoptosis and cell cycle arrest in both gastric cancer cells and hepatoma cells

Drug Des Devel Ther. 2018 Aug 20:12:2567-2575. doi: 10.2147/DDDT.S171675. eCollection 2018.

Abstract

Objectives: Calcineurin B (CNB) is a regulatory subunit of calcineurin, and it has antitumor activity. In this study, we aimed to investigate the effect of recombinant human calcineurin B (rhCNB) on the proliferation of gastric cancer cells and hepatoma cells both in vitro and in vivo.

Materials and methods: Cell viability and cell proliferation were detected by MTT and BrdU assay. Flow cytometry, Western blot and immunohistochemistry were performed to determine rhCNB-induced apoptosis and cell cycle arrest. The antitumor activities of rhCNB were observed in mice tumor models.

Results: We demonstrated that rhCNB inhibits the proliferation of gastric cancer cells and hepatoma cells both in vitro and in vivo. We showed that the inhibition of cell proliferation by rhCNB is associated with apoptosis and cell cycle arrest in both tumor cell lines. Furthermore, we indicated that rhCNB promotes p53 protein expression, a potent proapoptotic factor. Meanwhile, we also exhibited that rhCNB decreases the expression of both cyclin B1 and CDK1 proteins, two proteins associated with G2/M arrest.

Conclusion: Together, these findings suggest that rhCNB markedly inhibits tumor growth and provides guidance for its drug development.

Keywords: apoptosis; cell cycle arrest; p53; rhCNB; tumor cells.

MeSH terms

  • Animals
  • Antineoplastic Agents / pharmacology*
  • Apoptosis / drug effects*
  • CDC2 Protein Kinase / metabolism
  • Calcineurin / genetics
  • Calcineurin / pharmacology*
  • Carcinoma, Hepatocellular / drug therapy*
  • Carcinoma, Hepatocellular / metabolism
  • Carcinoma, Hepatocellular / pathology
  • Cell Proliferation / drug effects
  • Cyclin B1 / metabolism
  • G2 Phase Cell Cycle Checkpoints / drug effects*
  • Hep G2 Cells
  • Humans
  • Liver Neoplasms / drug therapy*
  • Liver Neoplasms / metabolism
  • Liver Neoplasms / pathology
  • Mice, Inbred BALB C
  • Mice, Nude
  • Protein Engineering / methods*
  • Recombinant Proteins / pharmacology
  • Signal Transduction / drug effects
  • Stomach Neoplasms / drug therapy*
  • Stomach Neoplasms / metabolism
  • Stomach Neoplasms / pathology
  • Tumor Suppressor Protein p53 / metabolism
  • Xenograft Model Antitumor Assays

Substances

  • Antineoplastic Agents
  • CCNB1 protein, human
  • Cyclin B1
  • Recombinant Proteins
  • TP53 protein, human
  • Tumor Suppressor Protein p53
  • CDC2 Protein Kinase
  • CDK1 protein, human
  • Calcineurin