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Betulinic acid decreases ER-negative breast cancer cell growth in vitro and in vivo: Role of Sp transcription factors and microRNA-27a:ZBTB10

Authors

  • Susanne U. Mertens-Talcott,

    Corresponding author
    1. Department of Nutrition and Food Science, Texas A&M University, College Station, Texas
    2. Department of Veterinary Physiology and Pharmacology, College of Veterinary Medicine, Texas A&M University, College Station, Texas
    3. Institute for Obesity Research and Program Evaluation, Texas A&M University, College Station, Texas
    • 1500 Research Parkway 220k, MS 2254, College Station, TX 77843.
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  • Giuliana D. Noratto,

    1. Department of Nutrition and Food Science, Texas A&M University, College Station, Texas
    2. Department of Veterinary Physiology and Pharmacology, College of Veterinary Medicine, Texas A&M University, College Station, Texas
    3. Institute for Obesity Research and Program Evaluation, Texas A&M University, College Station, Texas
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  • Xiangrong Li,

    1. Department of Nutrition and Food Science, Texas A&M University, College Station, Texas
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  • Gabriela Angel-Morales,

    1. Department of Nutrition and Food Science, Texas A&M University, College Station, Texas
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  • Michele C. Bertoldi,

    1. Department of Nutrition and Food Science, Texas A&M University, College Station, Texas
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  • Stephen Safe

    1. Department of Veterinary Physiology and Pharmacology, College of Veterinary Medicine, Texas A&M University, College Station, Texas
    2. Center for Environmental and Genetic Medicine, Texas A&M Health Science Center, Institute of Biosciences and Technology, Houston, Texas
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  • Conflict of interest: none.

Abstract

Betulinic acid (BA), a pentacyclic triterpenoid isolated from tree bark is cytotoxic to cancer cells. There is evidence that specificity proteins (Sps), such as Sp1, Sp3, and Sp4, are overexpressed in tumors and contribute to the proliferative and angiogenic phenotype associated with cancer cells. The objective of this study was to determine the efficacy of BA in decreasing the Sps expression and underlying mechanisms. Results show that BA decreased proliferation and induced apoptosis of estrogen-receptor-negative breast cancer MDA-MB-231 cells. The BA-induced Sp1, Sp3, and Sp4 downregulation was accompanied by increased zinc finger ZBTB10 expression, a putative Sp-repressor and decreased microRNA-27a levels, a microRNA involved in the regulation of ZBTB10. Similar results were observed in MDA-MB-231 cells transfected with ZBTB10 expression plasmid. BA induced cell cycle arrest in the G2/M phase and increased Myt-1 mRNA (a microRNA-27a target gene), which causes inhibition in G2/M by phosphorylation of cdc2. The effects of BA were reversed by transient transfection with a mimic of microRNA-27a. In nude mice with xenografted MDA-MB-231 cells, tumor size and weight were significantly decreased by BA treatment. In tumor tissue, ZBTB10 mRNA was increased while mRNA and protein of Sp1, Sp3 and Sp4, as well as mRNA of vascular endothelial growth factor receptor (VEGFR), survivin and microRNA-27a were decreased by BA. In lungs of xenografted mice, human β2-microglobulin mRNA was decreased in BA-treated animals. These results show that the anticancer effects of BA are at least in part based on interactions with the microRNA-27a-ZBTB10-Sp-axis causing increased cell death. © 2012 Wiley Periodicals, Inc.

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