ABSTRACT
cJun NH(2)-terminal kinase (JNK) signaling has been implicated in the developmental morphogenesis of epithelial organs. In this study, we employed a compound deletion of the murine Jnk1 and Jnk2 genes in the mammary gland to evaluate the requirement for these ubiquitously expressed genes in breast development and tumorigenesis. JNK1/2 was not required for breast epithelial cell proliferation or motility. However, JNK1/2 deficiency caused increased branching morphogenesis and defects in the clearance of lumenal epithelial cells. In the setting of breast cancer development, JNK1/2 deficiency significantly increased tumor formation. Together, these findings established that JNK signaling is required for normal mammary gland development and that it has a suppressive role in mammary tumorigenesis.
Subject(s)
MAP Kinase Signaling System/physiology , Mammary Glands, Animal/enzymology , Mammary Glands, Animal/growth & development , Mammary Neoplasms, Experimental/enzymology , Mitogen-Activated Protein Kinase 8/metabolism , Mitogen-Activated Protein Kinase 9/metabolism , Animals , Cell Growth Processes/physiology , Cell Movement/physiology , Female , Gene Expression , Mammary Neoplasms, Experimental/genetics , Mammary Neoplasms, Experimental/pathology , Mice , Mice, Inbred C57BL , Mice, Nude , Mice, Transgenic , Mitogen-Activated Protein Kinase 8/deficiency , Mitogen-Activated Protein Kinase 8/genetics , Mitogen-Activated Protein Kinase 9/deficiency , Mitogen-Activated Protein Kinase 9/geneticsABSTRACT
The c-Jun NH(2)-terminal kinase (JNK) signal transduction pathway causes increased gene expression mediated, in part, by members of the activating transcription factor protein (AP1) group. JNK is therefore implicated in the regulation of cell growth and cancer. To test the role of JNK in Ras-induced tumor formation, we examined the effect of compound ablation of the ubiquitously expressed genes Jnk1 plus Jnk2. We report that JNK is required for Ras-induced transformation of p53-deficient primary cells in vitro. Moreover, JNK is required for lung tumor development caused by mutational activation of the endogenous KRas gene in vivo. Together, these data establish that JNK plays a key role in Ras-induced tumorigenesis.
Subject(s)
JNK Mitogen-Activated Protein Kinases/metabolism , Lung Neoplasms/enzymology , Lung Neoplasms/pathology , Precancerous Conditions/enzymology , Precancerous Conditions/pathology , Proto-Oncogene Proteins p21(ras)/metabolism , Animals , Apoptosis , Cadherins/metabolism , Cell Proliferation , Cell Transformation, Neoplastic/pathology , Contact Inhibition , Fibroblasts/metabolism , Mice , Signal Transduction , Stress, Physiological , Tumor Stem Cell Assay , Tumor Suppressor Protein p53/metabolismABSTRACT
Mitogen-activated protein kinase (MAPK) cascades propagate a variety of cellular activities. Processive relay of signals through RAF-MEK-ERK modulates cell growth and proliferation. Signalling through this ERK cascade is frequently amplified in cancers, and drugs such as sorafenib (which is prescribed to treat renal and hepatic carcinomas) and PLX4720 (which targets melanomas) inhibit RAF kinases. Natural factors that influence ERK1/2 signalling include the second messenger cyclic AMP. However, the mechanisms underlying this cascade have been difficult to elucidate. We demonstrate that the A-kinase-anchoring protein AKAP-Lbc and the scaffolding protein kinase suppressor of Ras (KSR-1) form the core of a signalling network that efficiently relay signals from RAF, through MEK, and on to ERK1/2. AKAP-Lbc functions as an enhancer of ERK signalling by securing RAF in the vicinity of MEK1 and synchronizing protein kinase A (PKA)-mediated phosphorylation of Ser 838 on KSR-1. This offers mechanistic insight into cAMP-responsive control of ERK signalling events.
Subject(s)
A Kinase Anchor Proteins/metabolism , Cyclic AMP/metabolism , MAP Kinase Signaling System , Mitogen-Activated Protein Kinase 1/metabolism , Mitogen-Activated Protein Kinase 3/metabolism , Proto-Oncogene Proteins/metabolism , Amino Acid Sequence , Animals , Cell Line , Cyclic AMP-Dependent Protein Kinases/metabolism , Humans , Mice , Minor Histocompatibility Antigens , Molecular Sequence Data , Protein Kinases/chemistry , Protein Kinases/metabolism , Sequence AlignmentABSTRACT
The cJun NH2-terminal kinase (JNK) signal transduction pathway has been implicated in mammary carcinogenesis. To test the role of JNK, we examined the effect of ablation of the Jnk1 and Jnk2 genes in a Trp53-dependent model of breast cancer using BALB/c mice. We detected no defects in mammary gland development in virgin mice or during lactation and involution in control studies of Jnk1(-/-) and Jnk2(-/-) mice. In a Trp53(-/+) genetic background, mammary carcinomas were detected in 43% of control mice, 70% of Jnk1(-/-) mice, and 53% of Jnk2(-/-) mice. These data indicate that JNK1 and JNK2 are not essential for mammary carcinoma development in the Trp53(-/+) BALB/c model of breast cancer. In contrast, this analysis suggests that JNK may partially contribute to tumor suppression. This conclusion is consistent with the finding that tumor-free survival of JNK-deficient Trp53(-/+) mice was significantly reduced compared with control Trp53(-/+) mice. We conclude that JNK1 and JNK2 can act as suppressors of mammary tumor development.
Subject(s)
Breast Neoplasms/enzymology , Mitogen-Activated Protein Kinase 8/metabolism , Mitogen-Activated Protein Kinase 9/metabolism , Tumor Suppressor Protein p53/metabolism , Animals , Breast Neoplasms/metabolism , Breast Neoplasms/pathology , Disease Models, Animal , Female , Mammary Glands, Animal/enzymology , Mammary Glands, Animal/growth & development , Mice , Mice, Inbred BALB C , Mitogen-Activated Protein Kinase 8/deficiency , Mitogen-Activated Protein Kinase 9/deficiency , Survival Analysis , Tumor Suppressor Protein p53/deficiencyABSTRACT
The activity of the ERK has complex spatial and temporal dynamics that are important for the specificity of downstream effects. However, current biochemical techniques do not allow for the measurement of ERK signaling with fine spatiotemporal resolution. We developed a genetically encoded, FRET-based sensor of ERK activity (the extracellular signal-regulated kinase activity reporter, EKAR), optimized for signal-to-noise ratio and fluorescence lifetime imaging. EKAR selectively and reversibly reported ERK activation in HEK293 cells after epidermal growth factor stimulation. EKAR signals were correlated with ERK phosphorylation, required ERK activity, and did not report the activities of JNK or p38. EKAR reported ERK activation in the dendrites and nucleus of hippocampal pyramidal neurons in brain slices after theta-burst stimuli or trains of back-propagating action potentials. EKAR therefore permits the measurement of spatiotemporal ERK signaling dynamics in living cells, including in neuronal compartments in intact tissues.
Subject(s)
Extracellular Signal-Regulated MAP Kinases/metabolism , Fluorescence Resonance Energy Transfer/methods , Fluorescent Dyes , MAP Kinase Signaling System , Pyramidal Cells/enzymology , Action Potentials/physiology , Animals , Artifacts , Bacterial Proteins/genetics , Cell Line , Dendrites/enzymology , Endothelial Cells/cytology , Endothelial Cells/enzymology , Enzyme Activation , Extracellular Signal-Regulated MAP Kinases/genetics , Genes, Reporter , Green Fluorescent Proteins/genetics , Hippocampus/cytology , Humans , Kidney/cytology , Luminescent Proteins/genetics , Organ Culture Techniques , Patch-Clamp Techniques , RatsABSTRACT
It is established that p38 MAPK can negatively regulate tumorigenesis, but the mechanism is incompletely understood. A new study in this issue of Cancer Cell shows that p38 MAP kinase plays a selective role in tumor initiation mediated by oxidative stress.
