Selective induction of apoptosis in mutant p53 premalignant and malignant cancer cells by PRIMA-1 through the c-Jun-NH2-kinase pathway.

Abstract

PRIMA-1 (p53 reactivation and induction of massive apoptosis) is a chemical compound that was originally identified as a selective mutant p53-dependent growth suppressor by screening a library of low-molecular-weight compounds. However, its mechanism of action is unknown. In this study, we examined toxicity of PRIMA-1 to three premalignant human colorectal adenoma cell lines (RG/C2, BR/C1, and AA/C1) and four colorectal carcinoma cell lines (DLD-1, SW480, LOVO, and HCT116) and its mechanism of action. It selectively induced apoptosis only in the mutant p53 premalignant and malignant colon cell lines, but was not toxic to the wild-type p53 premalignant and malignant colon cell lines. Using stable transfectants of temperature-sensitive p53 mutant Ala(143) in null p53 H1299 lung cancer cells, we found that PRIMA-1 induced significantly more apoptosis in cells with mutant p53 conformation (37 degrees C) than the wild-type p53 conformation (32.5 degrees C). Cell cycle analysis indicated that its inhibition of cell growth was correlated with induction of G(2) arrest. Western blot analysis showed PRIMA-1 increased p21 and GADD45 expression selectively in the mutant p53 cells. However, Fas, Bcl-2 family proteins, and caspases were not involved in PRIMA-1-induced cell death. The c-Jun-NH(2)-kinase (JNK) inhibitor SP 600125, but not p38 mitogen-activated protein kinase inhibitor SB 203580 or extracellular signal-regulated kinase inhibitor PD 98059, blocked PRIMA-1-induced apoptosis. Transfection with a dominant-negative phosphorylation mutant JNK, but not a dominant-negative p38 or wild-type JNK, inhibited PRIMA-1-induced cell death, suggesting that the JNK pathway plays an important role in PRIMA-1-induced apoptosis. PRIMA-1 is a highly selective small molecule toxic to p53 mutant cells and may serve as a prototype for the development of new p53-targeting agents for therapy of premalignant and malignant cells.

• Authors: Yin Li, Yuehua Mao, Paul W Brandt-Rauf, Ann C Williams, Robert L Fine
• Journal: Molecular cancer therapeutics (Mol Cancer Ther) Vol. 4 Issue 6 Pg. 901-9 (Jun 2005) ISSN: 1535-7163 [Print] United States
• PMID: 15956247 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Aza Compounds
• Bridged Bicyclo Compounds, Heterocyclic
• Protein Kinase Inhibitors
• RNA, Messenger
• Tumor Suppressor Protein p53
• JNK Mitogen-Activated Protein Kinases
• 2,2-bis(hydroxymethyl)-1-azabicyclo(2,2,2,)octan-3-one

Topics

• Apoptosis (drug effects)
• Aza Compounds (pharmacology)
• Bridged Bicyclo Compounds, Heterocyclic (pharmacology)
• Cell Line
• Cell Transformation, Neoplastic (genetics, metabolism, pathology)
• Colonic Neoplasms (metabolism, pathology)
• Humans
• JNK Mitogen-Activated Protein Kinases (antagonists & inhibitors, genetics, metabolism)
• Lung Neoplasms (metabolism, pathology)
• Neoplasms (genetics, metabolism, pathology)
• Precancerous Conditions (genetics, metabolism)
• Protein Kinase Inhibitors (pharmacology)
• RNA, Messenger (genetics, metabolism)
• Signal Transduction (drug effects)
• Temperature
• Tumor Suppressor Protein p53 (genetics)