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Valproic acid induces p21 and topoisomerase-II (α/β) expression and synergistically enhances etoposide cytotoxicity in human glioblastoma cell lines

  • Lab. Investigation-Human/Animal Tissue
  • Published:
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Abstract

Object

Etoposide, a topoisomerase-II inhibitor promotes DNA damage and apoptosis of cancer cells. In this study, we have examined the ability of the histone deacetylase inhibitor, valproic acid (VPA) to modulate gene expression and sensitize glioblastoma cell lines to the cytotoxic effects of etoposide in vitro.

Methods

The effect of VPA and etoposide alone or a combination of the two drugs on the growth of three different glioblastoma cell lines (U87, LN18, and U251) were measured by MTT assays. Drug treated cells were analyzed for their cell cycle profile, gene expression, differentiation status, and induction of apoptosis by flow-cytometry, western blotting, immunofluorescence assays, and caspase activity measurements.

Results

We observed that while VPA and etoposide independently inhibited the growth of U87, U251, and LN18 cells, exposure of tumor cells to both drugs significantly enhanced the cytotoxicity of etoposide in all cell lines. VPA promoted a G1 accumulation of U87, while an increase in the G2/M population of U251 and LN18 cells was observed upon exposure to the drug. Treatment with etoposide resulted in a G2/M arrest of U87, U251, and LN18 cells, whereas, exposure to both drugs increased the fraction of cells with a G2/M and sub-G1 DNA content. Further, VPA and not etoposide, promoted acetylation of histone H4 and induced the expression of the cyclin-dependent kinase inhibitor (CDKI), p21/WAF1. VPA also up-regulated the expression of the α and β isoforms of topoisomerase-II, as well as the glial differentiation marker, glial fibrillary acidic protein. Finally, a significant increase in caspase-3 activity and apoptosis was observed in the presence of both VPA and etoposide compared to either agent alone.

Conclusion

Our study demonstrates that VPA sensitizes U87, U251, and LN18 cells to the cytotoxic effects of etoposide in vitro by inducing differentiation and up-regulating the expression of p21/WAF1 and both isoforms of topoisomerase-II.

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Acknowledgments

We thank Drs. Joya Chandra, Peter Zage, Neeta Kang, and Jason Long for their technical help and critical reading of the manuscript. This work was supported in part by funds from The Children’s Brain Tumor Foundation to VG.

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Authors and Affiliations

  1. Department of Pediatrics, Unit 853, University of Texas, M.D. Anderson Cancer Center, 1515 Holcombe Blvd, Houston, TX, 77030, USA

    Chandra M. Das, Dolly Aguilera, Hernan Vasquez, Preethi Prasad, Johannes E. Wolff & Vidya Gopalakrishnan

  2. Department of Biostatistics and Applied Math, University of Texas, M.D. Anderson Cancer Center, 1515 Holcombe Blvd, Houston, TX, 77030, USA

    Ming Zhang

  3. University of Texas Graduate School of Biomedical Sciences, 1515 Holcombe Blvd, Houston, TX, 77030, USA

    Johannes E. Wolff & Vidya Gopalakrishnan

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  1. Chandra M. Das
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Correspondence to Vidya Gopalakrishnan.

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Das, C.M., Aguilera, D., Vasquez, H. et al. Valproic acid induces p21 and topoisomerase-II (α/β) expression and synergistically enhances etoposide cytotoxicity in human glioblastoma cell lines. J Neurooncol 85, 159–170 (2007). https://doi.org/10.1007/s11060-007-9402-7

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