Inhibition of MDA-MB-231 breast cancer cell proliferation and tumor growth by apigenin through induction of G2/M arrest and histone H3 acetylation-mediated p21 WAF1/CIP1 expression

Environ Toxicol. 2017 Feb;32(2):434-444. doi: 10.1002/tox.22247. Epub 2016 Feb 12.

Abstract

Apigenin (4',5,7-trihydroxyflavone), a flavonoid commonly found in fruits and vegetables, has anticancer properties in various malignant cancer cells. However, the molecular basis of the anticancer effect remains to be elucidated. In this study, we investigated the cellular mechanisms underlying the induction of cell cycle arrest by apigenin. Our results showed that apigenin at the nonapoptotic induction concentration inhibited cell proliferation and induced cell cycle arrest at the G2/M phase in the MDA-MB-231 breast cancer cell line. Immunoblot analysis indicated that apigenin suppressed the expression of cyclin A, cyclin B, and cyclin-dependent kinase-1 (CDK1), which control the G2-to-M phase transition in the cell cycle. In addition, apigenin upregulated p21WAF1/CIP1 and increased the interaction of p21WAF1/CIP1 with proliferating cell nuclear antigen (PCNA), which inhibits cell cycle progression. Furthermore, apigenin significantly inhibited histone deacetylase (HDAC) activity and induced histone H3 acetylation. The subsequent chromatin immunoprecipitation (ChIP) assay indicated that apigenin increased acetylation of histone H3 in the p21WAF1/CIP1 promoter region, resulting in the increase of p21WAF1/CIP1 transcription. In a tumor xenograft model, apigenin effectively delayed tumor growth. In these apigenin-treated tumors, we also observed reductions in the levels of cyclin A and cyclin B and increases in the levels of p21WAF1/CIP1 and acetylated histone H3. These findings demonstrate for the first time that apigenin can be used in breast cancer prevention and treatment through epigenetic regulation. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 434-444, 2017.

Keywords: apigenin; breast cancer; cell cycle arrest; histone H3 acetylation; p21WAF1/CIP1.

MeSH terms

  • Acetylation
  • Animals
  • Antineoplastic Agents, Phytogenic / pharmacology*
  • Apigenin / pharmacology*
  • Breast Neoplasms / drug therapy
  • Cell Cycle Checkpoints / drug effects*
  • Cell Division / drug effects*
  • Cell Line, Tumor
  • Cell Proliferation / drug effects*
  • Cyclin-Dependent Kinase Inhibitor p21 / metabolism*
  • Epigenesis, Genetic / drug effects
  • Female
  • G2 Phase / drug effects*
  • Gene Expression Regulation, Neoplastic / drug effects
  • Histone Deacetylases / analysis
  • Histone Deacetylases / metabolism
  • Histones / metabolism*
  • Humans
  • Mice
  • Mice, Inbred BALB C
  • Xenograft Model Antitumor Assays

Substances

  • Antineoplastic Agents, Phytogenic
  • CDKN1A protein, human
  • Cyclin-Dependent Kinase Inhibitor p21
  • Histones
  • Apigenin
  • Histone Deacetylases