Characterization of p21Ras-mediated apoptosis induced by protein kinase C inhibition and application to human tumor cell lines.

Abstract

Suppression of PKC activity can selectively induce apoptosis in cells expressing a constitutively activated p21Ras protein. We demonstrate that continued expression of p21Ras activity is required in PKC-mediated apoptosis because farnesyltransferase inhibitors abrogated the loss of viability in p21Ras-transformed cells occurring following PKC inhibition. Studies utilizing gene transfer or viral vectors demonstrate that transient expression of oncogenic p21Ras activity is sufficient for induction of apoptosis by PKC inhibition, whereas physiologic activation of p21Ras by growth factor is not sufficient to induce apoptosis. Mechanistically, the p21Ras-mediated apoptosis induced by PKC inhibition is dependent upon mitochondrial dysregulation, with a concurrent loss of mitochondrial membrane potential (psim). Cyclosporine A, which prevented the loss of psim, also inhibited HMG-induced DNA fragmentation in cells expressing an activated p21Ras. Induction of apoptosis by PKC inhibition in human tumors with oncogenic p21Ras mutations was demonstrated. Inhibition of PKC caused increased apoptosis in MIA-PaCa-2, a human pancreatic tumor line containing a mutated Ki-ras allele, when compared to HS766T, a human pancreatic tumor line with normal Ki-ras alleles. Furthermore, PKC inhibition induced apoptosis in HCT116, a human colorectal tumor line containing an oncogenic Ki-ras allele but not in a subline (Hke3) in which the mutated Ki-ras allele had been disrupted. The PKC inhibitor 1-O-hexadecyl-2-O-methyl-rac-glycerol (HMG), significantly reduced p21Ras-mediated tumor growth in vivo in a nude mouse MIA-PaCa-2 xenograft model. Collectively these studies suggest the therapeutic feasibility of targeting PKC activity in tumors expressing an activated p21Ras oncoprotein.

• Authors: James S Liou, James S Chen, Douglas V Faller
• Journal: Journal of cellular physiology (J Cell Physiol) Vol. 198 Issue 2 Pg. 277-94 (Feb 2004) ISSN: 0021-9541 [Print] United States
• PMID: 14603530 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)
• Copyright: Copyright 2003 Wiley-Liss, Inc.

Chemical References

• Enzyme Inhibitors
• Glyceryl Ethers
• 1-O-hexadecyl-2-O-methylglycerol
• Protein Kinase C
• HRAS protein, human
• Proto-Oncogene Proteins p21(ras)

Topics

• Animals
• Apoptosis (drug effects, physiology)
• CHO Cells
• Cell Line, Tumor (drug effects)
• Cricetinae
• Cricetulus
• Enzyme Inhibitors (pharmacology)
• Genes, ras (physiology)
• Glyceryl Ethers (pharmacology)
• Humans
• Jurkat Cells
• Membrane Potentials (drug effects)
• Mice
• Mitochondria (pathology, physiology)
• Mutation
• NIH 3T3 Cells
• Protein Kinase C (antagonists & inhibitors)
• Proto-Oncogene Proteins p21(ras) (physiology)