Differential radiosensitizing effect of valproic acid in differentiation versus self-renewal promoting culture conditions

Int J Radiat Oncol Biol Phys. 2010 Mar 1;76(3):889-95. doi: 10.1016/j.ijrobp.2009.09.052.

Abstract

Purpose: It has been shown that valproic acid (VA) enhances the proliferation and self-renewal of normal hematopoietic stem cells and that breast cancer stem/progenitor cells can be resistant to radiation. From these data, we hypothesized that VA would fail to radiosensitize breast cancer stem/progenitor cells grown to three-dimensional (3D) mammospheres.

Methods and materials: We used the MCF7 breast cancer cell line grown under stem cell-promoting culture conditions (3D mammosphere) and standard nonstem cell monolayer culture conditions (two-dimensional) to examine the effect of pretreatment with VA on radiation sensitivity in clonogenic survival assays and on the expression of embryonic stem cell transcription factors.

Results: 3D-cultured MCF-7 cells expressed higher levels of Oct4, Nanog, and Sox2. The 3D passage enriched self-renewal and increased radioresistance in the 3D mammosphere formation assays. VA radiosensitized adherent cells but radioprotected 3D cells in single-fraction clonogenic assays. Moreover, fractionated radiation sensitized VA-treated adherent MCF7 cells but did not have a significant effect on VA-treated single cells grown to mammospheres.

Conclusion: We have concluded that VA might preferentially radiosensitize differentiated cells compared with those expressing stem cell surrogates and that stem cell-promoting culture is a useful tool for in vitro evaluation of novel cancer therapeutic agents and radiosensitizers.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Culture Techniques / methods
  • Cell Differentiation / drug effects
  • Cell Line, Tumor
  • Cell Proliferation
  • Homeodomain Proteins / metabolism
  • Humans
  • Nanog Homeobox Protein
  • Neoplastic Stem Cells / cytology
  • Neoplastic Stem Cells / drug effects
  • Neoplastic Stem Cells / metabolism
  • Neoplastic Stem Cells / radiation effects*
  • Octamer Transcription Factor-3 / metabolism
  • Radiation Tolerance / drug effects*
  • Radiation Tolerance / physiology
  • Radiation-Protective Agents / pharmacology
  • Radiation-Sensitizing Agents / pharmacology*
  • SOXB1 Transcription Factors / metabolism
  • Spheroids, Cellular / drug effects
  • Spheroids, Cellular / metabolism
  • Spheroids, Cellular / pathology
  • Spheroids, Cellular / radiation effects
  • Tumor Stem Cell Assay / methods
  • Valproic Acid / pharmacology*

Substances

  • Homeodomain Proteins
  • NANOG protein, human
  • Nanog Homeobox Protein
  • Octamer Transcription Factor-3
  • Radiation-Protective Agents
  • Radiation-Sensitizing Agents
  • SOX2 protein, human
  • SOXB1 Transcription Factors
  • Valproic Acid