Growth inhibition induced by Ro 31-8220 and Calphostin C in human glioblastoma cell lines is associated with apoptosis and inhibition of CDC2 kinase

Martin Begemann, Sharafadeen A. Kashimawo, Ruth M. Lunn, Thomas Delohery, Yu Jeong A. Choi, Susan Kim, Daniel F. Heitjan, Regina M. Santella, Peter B. Schiff, Jeffrey N. Bruce, I. Bernard Weinstein

Research output: Contribution to journalArticle

29 Scopus citations

Abstract

Protein kinase C (PKC) is a central component in signal transduction and growth control and might be an appropriate target for the chemotherapy of human brain tumors. This study demonstrates that the staurosporine derivative Ro 31-8220, a potent PKC inhibitor, inhibited the growth of 7 human brain tumor cell lines with an IC50 of about 2 μM. Calphostin C, a structurally unrelated PKC inhibitor, inhibited the growth of two of these cell lines with an IC50 of about 100 to 300 nM. Drug withdrawal and clonogenicity assays indicated that the growth inhibition by both of these compounds was irreversible. Morphologic studies, DNA fragmentation studies and flow cytometric assays showed that the treated glioblastoma cells underwent apoptosis. Treatment of glioblastoma cells with Ro 31-8220 lend to a rapid decline in the level of the anti-apoptosis protein bcl-2. At least three of the glioblastoma cell lines carried mutant p53 alleles with missense mutations in the DNA binding domain of p53. Therefore, the induction of apoptosis in these cell lines occurred through a p53-independent mechanism. Furthermore treatment of these glioblastoma cell lines with Ro 31-8220 or calphostin C led to an increase of cells in the G2-M phase of the cell cycle. This correlated with a decrease in CDC2-associated histone H1 kinase activity as well as a decrease in the level of the CDC2 protein as shown by immunoblotting. When added to subcellullar assays Ro 31-8220 markedly inhibited CDC2 histone H1 kinase activity with an IC50 of 100 nM, but calphostin C directly inhibited this kinase activity only at very high concentrations (above 100 μM). Thus these compounds inhibit the growth of glioblastoma cells through novel mechanisms. Ro 31-8220, in particular, might be a useful agent for the treatment of human brain tumors.

Original languageEnglish (US)
Pages (from-to)3139-3152
Number of pages14
JournalAnticancer Research
Volume18
Issue number5 A
StatePublished - Sep 1 1998

Keywords

  • Bcl-2
  • Chemotherapy
  • PKC
  • p53

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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    Begemann, M., Kashimawo, S. A., Lunn, R. M., Delohery, T., Choi, Y. J. A., Kim, S., Heitjan, D. F., Santella, R. M., Schiff, P. B., Bruce, J. N., & Weinstein, I. B. (1998). Growth inhibition induced by Ro 31-8220 and Calphostin C in human glioblastoma cell lines is associated with apoptosis and inhibition of CDC2 kinase. Anticancer Research, 18(5 A), 3139-3152.