10-formyltetrahydrofolate dehydrogenase-induced c-Jun-NH2-kinase pathways diverge at the c-Jun-NH2-kinase substrate level in cells with different p53 status.

Abstract

10-Formyltetrahydrofolate dehydrogenase (FDH) suppresses cancer cell proliferation through p53-dependent apoptosis but also induces strong cytotoxicity in p53-deficient prostate cells. In the present study, we have shown that FDH induces apoptosis in PC-3 prostate cells through simultaneous activation of the c-Jun-NH(2)-kinase (JNK) and extracellular signal-regulated kinase (ERK) pathways with JNK phosphorylating c-Jun and ERK1/2 phosphorylating Elk-1. The JNK1/2 inhibitor SP600125 or ERK1/2 inhibitor PD98059 prevented phosphorylation of c-Jun and Elk-1, correspondingly and partially protected PC-3 cells from FDH-induced cytotoxicity. Combination of the two inhibitors produced an additive effect. The contribution from the JNK cascade to FDH-induced apoptosis was significantly stronger than from the ERK pathway. siRNA knockdown of JNK1/2 or "turning off" the downstream target c-Jun by either siRNA or expression of the dominant-negative c-Jun mutant, TAM67, rescued PC-3 cells from FDH-induced apoptosis. The pull-down assays on immobilized c-Jun showed that c-Jun is directly phosphorylated by JNK2 in FDH-expressing cells. Interestingly, the FDH-induced apoptosis in p53-proficient A549 cells also proceeds through activation of JNK1/2, but the down-stream target for JNK2 is p53 instead of c-Jun. Furthermore, in A549 cells, FDH activates caspase 9, whereas in PC-3 cells, it activates caspase 8. Our studies indicate that the JNK pathways are common downstream mechanisms of FDH-induced cytotoxicity in different cell types, whereas the end point target in the cascade is cell type specific. JNK activation in response to FDH was inhibited by high supplementation of reduced folate leucovorin, further indicating a functional connection between folate metabolism and mitogen-activated protein kinase pathways.

• Authors: Sampa Ghose, Natalia V Oleinik, Natalia I Krupenko, Sergey A Krupenko
• Journal: Molecular cancer research : MCR (Mol Cancer Res) Vol. 7 Issue 1 Pg. 99-107 (Jan 2009) ISSN: 1541-7786 [Print] United States
• PMID: 19147541 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Chemical References

• RNA, Neoplasm
• RNA, Small Interfering
• Oxidoreductases Acting on CH-NH Group Donors
• formyltetrahydrofolate dehydrogenase
• Extracellular Signal-Regulated MAP Kinases
• JNK Mitogen-Activated Protein Kinases
• p38 Mitogen-Activated Protein Kinases

Topics

• Adenocarcinoma (enzymology, genetics)
• Apoptosis
• Cell Line, Tumor
• Extracellular Signal-Regulated MAP Kinases (metabolism)
• Genes, p53
• Genetic Variation
• Humans
• JNK Mitogen-Activated Protein Kinases (genetics, metabolism)
• Male
• Oxidoreductases Acting on CH-NH Group Donors (metabolism)
• Phosphorylation
• Prostatic Neoplasms (enzymology, genetics)
• RNA Interference
• RNA, Neoplasm (genetics)
• RNA, Small Interfering (genetics)
• Substrate Specificity
• Transfection
• p38 Mitogen-Activated Protein Kinases (genetics, metabolism)