Astaxanthin down-regulates Rad51 expression via inactivation of AKT kinase to enhance mitomycin C-induced cytotoxicity in human non-small cell lung cancer cells

Biochem Pharmacol. 2016 Apr 1:105:91-100. doi: 10.1016/j.bcp.2016.02.016. Epub 2016 Feb 24.

Abstract

Astaxanthin has been demonstrated to exhibit a wide range of beneficial effects, including anti-inflammatory and anti-cancer properties. However, the molecular mechanism of astaxanthin-induced cytotoxicity in non-small cell lung cancer (NSCLC) cells has not been identified. Rad51 plays a central role in homologous recombination, and studies show that chemo-resistant carcinomas exhibit high levels of Rad51 expression. In this study, astaxanthin treatment inhibited cell viability and proliferation of two NSCLC cells, A549 and H1703. Astaxanthin treatment (2.5-20 μM) decreased Rad51 expression and phospho-AKT(Ser473) protein level in a time and dose-dependent manner. Furthermore, expression of constitutively active AKT (AKT-CA) vector rescued the decreased Rad51 mRNA and protein levels in astaxanthin-treated NSCLC cells. Combined treatment with phosphatidylinositol 3-kinase (PI3K) inhibitors (LY294002 or wortmannin) further decreased the Rad51 expression in astaxanthin-exposed A549 and H1703 cells. Knockdown of Rad51 expression by transfection with si-Rad51 RNA or cotreatment with LY294002 further enhanced the cytotoxicity and cell growth inhibition of astaxanthin. Additionally, mitomycin C (MMC) as an anti-tumor antibiotic is widely used in clinical NSCLC chemotherapy. Combination of MMC and astaxanthin synergistically resulted in cytotoxicity and cell growth inhibition in NSCLC cells, accompanied with reduced phospho-AKT(Ser473) level and Rad51 expression. Overexpression of AKT-CA or Flag-tagged Rad51 reversed the astaxanthin and MMC-induced synergistic cytotoxicity. In contrast, pretreatment with LY294002 further decreased the cell viability in astaxanthin and MMC co-treated cells. In conclusion, astaxanthin enhances MMC-induced cytotoxicity by decreasing Rad51 expression and AKT activation. These findings may provide rationale to combine astaxanthin with MMC for the treatment of NSCLC.

Keywords: AKT; Actinomycin D (PubChem CID: 2019); Astaxanthin; Astaxanthin (PubChem CID: 5281224); Crystal violet (PubChem CID: 11057); Cycloheximide (PubChem CID: 6197); LY294002 (PubChem CID: 3973); Leupeptin (PubChem CID: 72429); MG132 (PubChem CID: 462382); Mitomycin C; Mitomycin C (PubChem CID: 5746); Non-small cell lung cancer; Penicillin (PubChem CID: 5904); Sodium bicarbonate (PubChem CID: 516892); Streptomycin (PubChem CID: 19649); Trypan blue (PubChem CID: 9562061); Wortmannin (PubChem CID: 312145); l-Glutamine (PubChem CID: 5961).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibiotics, Antineoplastic / toxicity
  • Carcinoma, Non-Small-Cell Lung / drug therapy
  • Carcinoma, Non-Small-Cell Lung / metabolism*
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Cell Survival / physiology
  • Dose-Response Relationship, Drug
  • Down-Regulation / drug effects
  • Down-Regulation / physiology
  • Humans
  • Lung Neoplasms / drug therapy
  • Lung Neoplasms / metabolism*
  • Mitomycin / therapeutic use
  • Mitomycin / toxicity*
  • Oncogene Protein v-akt / antagonists & inhibitors
  • Oncogene Protein v-akt / metabolism*
  • Rad51 Recombinase / antagonists & inhibitors
  • Rad51 Recombinase / biosynthesis*
  • Xanthophylls / toxicity

Substances

  • Antibiotics, Antineoplastic
  • Xanthophylls
  • Mitomycin
  • astaxanthine
  • Oncogene Protein v-akt
  • RAD51 protein, human
  • Rad51 Recombinase