Withaferin A inhibits experimental epithelial-mesenchymal transition in MCF-10A cells and suppresses vimentin protein level in vivo in breast tumors

• Published in: Molecular carcinogenesis Vol. 54; no. 6; pp. 417 - 429
• Main Authors: Lee, Joomin, Hahm, Eun-Ryeong, Marcus, Adam I., Singh, Shivendra V.
• Format: Journal Article
• Language: English
• Published: United States Blackwell Publishing Ltd 01.06.2015; Wiley Subscription Services, Inc
• Subjects: Angiogenesis; Animals; Antineoplastic Agents, Phytogenic - pharmacology; Antineoplastic Agents, Phytogenic - therapeutic use; Apoptosis - drug effects; Breast - drug effects; Breast - pathology; Breast cancer; Breast Neoplasms - drug therapy; Breast Neoplasms - metabolism; Breast Neoplasms - pathology; Cell Line, Tumor; Cell Movement - drug effects; EMT; Epithelial-Mesenchymal Transition - drug effects; Female; Herbal medicine; Humans; Mice, Nude; Proteins; Transforming Growth Factor beta - metabolism; Tumor Necrosis Factor-alpha - metabolism; vimentin; Vimentin - analysis; Vimentin - metabolism; withaferin A; Withanolides - pharmacology; Withanolides - therapeutic use; breast cancer; vimentin; withaferin A; EMT
• ISSN: 0899-1987; 1098-2744
• DOI: 10.1002/mc.22110

We have shown previously that withaferin A (WA), a bioactive component of the medicinal plant Withania somnifera, inhibits growth of cultured and xenografted human breast cancer cells and prevents breast cancer development and pulmonary metastasis incidence in a transgenic mouse model. The present study was undertaken to determine if the anticancer effect of WA involved inhibition of epithelial–mesenchymal transition (EMT). Experimental EMT induced by exposure of MCF‐10A cells to tumor necrosis factor‐α (TNF‐α) and transforming growth factor‐β1 (TGF‐β) was partially reversed by treatment with WA but not by its structural analogs withanone or withanolide A. Combined TNF‐α and TGF‐β treatments conferred partial protection against MCF‐10A cell migration inhibition by WA. Inhibition of TNF‐α and TGF‐β‐induced MCF‐10A cell migration by WA exposure was modestly attenuated by knockdown of E‐cadherin protein. MCF‐7 and MDA‐MB‐231 cells exposed to WA exhibited sustained (MCF‐7) or transient (MDA‐MB‐231) induction of E‐cadherin protein. On the other hand, the level of vimentin protein was increased markedly after 24 h treatment of MDA‐MB‐231 cells with WA. WA‐induced apoptosis was not affected by vimentin protein knockdown in MDA‐MB‐231 cells. Protein level of vimentin was significantly lower in the MDA‐MB‐231 xenografts as well as in MMTV‐neu tumors from WA‐treated mice compared with controls. The major conclusions of the present study are that (a) WA treatment inhibits experimental EMT in MCF‐10A cells, and (b) mammary cancer growth inhibition by WA administration is associated with suppression of vimentin protein expression in vivo. © 2013 Wiley Periodicals, Inc.