Metabolic alterations in mammary cancer prevention by withaferin A in a clinically relevant mouse model

J Natl Cancer Inst. 2013 Aug 7;105(15):1111-22. doi: 10.1093/jnci/djt153. Epub 2013 Jul 2.

Abstract

Background: Efficacy of withaferin A (WA), an Ayurvedic medicine constituent, for prevention of mammary cancer and its associated mechanisms were investigated using mouse mammary tumor virus-neu (MMTV-neu) transgenic model.

Methods: Incidence and burden of mammary cancer and pulmonary metastasis were scored in female MMTV-neu mice after 28 weeks of intraperitoneal administration with 100 µg WA (three times/week) (n = 32) or vehicle (n = 29). Mechanisms underlying mammary cancer prevention by WA were investigated by determination of tumor cell proliferation, apoptosis, metabolomics, and proteomics using plasma and/or tumor tissues. Spectrophotometric assays were performed to determine activities of complex III and complex IV. All statistical tests were two-sided.

Results: WA administration resulted in a statistically significant decrease in macroscopic mammary tumor size, microscopic mammary tumor area, and the incidence of pulmonary metastasis. For example, the mean area of invasive cancer was lower by 95.14% in the WA treatment group compared with the control group (mean = 3.10 vs 63.77 mm2, respectively; difference = -60.67 mm2; 95% confidence interval = -122.50 to 1.13 mm2; P = .0536). Mammary cancer prevention by WA treatment was associated with increased apoptosis, inhibition of complex III activity, and reduced levels of glycolysis intermediates. Proteomics confirmed downregulation of many glycolysis-related proteins in the tumor of WA-treated mice compared with control, including M2-type pyruvate kinase, phospho glycerate kinase, and fructose-bisphosphate aldolase A isoform 2.

Conclusions: This study reveals suppression of glycolysis in WA-mediated mammary cancer prevention in a clinically relevant mouse model.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antineoplastic Agents / pharmacology*
  • Apoptosis / drug effects
  • Cluster Analysis
  • Electrophoresis, Gel, Two-Dimensional
  • Female
  • Glycolysis / drug effects
  • Immunohistochemistry
  • In Situ Nick-End Labeling
  • Lung Neoplasms / prevention & control*
  • Lung Neoplasms / secondary
  • Mammary Neoplasms, Experimental / metabolism*
  • Mammary Neoplasms, Experimental / prevention & control*
  • Mammary Neoplasms, Experimental / virology
  • Mammary Tumor Virus, Mouse
  • Medicine, Ayurvedic
  • Mice
  • Mice, Transgenic
  • Random Allocation
  • Withanolides / pharmacology*

Substances

  • Antineoplastic Agents
  • Withanolides
  • withaferin A