The p38 SAPK pathway is required for Ha-ras induced in vitro invasion of NIH3T3 cells

Behren, Andreas; Binder, Konrad; Vucelic, Goran; Herberhold, Stephan; Hirt, Bernhard; Loewenheim, Hubert; Preyer, S; Zenner, H. P.; Simon, Christian

doi:10.1016/j.yexcr.2004.10.004

Cited by 16 publications

(15 citation statements)

References 35 publications

(53 reference statements)

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“…Binding of uPA with its receptor uPAR activates downstream signaling molecules through a number of pathways, including the mitogenactivated protein kinases and signal transducer and activator of transcription (Stat) pathways (19,(21)(22)(23)(24). Ample evidence suggests that enhanced signal transduction via the binding of uPA to uPAR contributes to the malignant phenotype of many cancers, and the particular nature of the response may be specific to cell type (16, 19, 24 -25).…”

mentioning

confidence: 99%

RNA Interference-directed Knockdown of Urokinase Plasminogen Activator and Urokinase Plasminogen Activator Receptor Inhibits Prostate Cancer Cell Invasion, Survival, and Tumorigenicity in Vivo

Pulukuri

Gondi

Lakka

et al. 2005

Journal of Biological Chemistry

243

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The invasive ability of tumor cells plays a key role in prostate cancer metastasis and is a major cause of treatment failure. Urokinase plasminogen activator-(uPA) and its receptor (uPAR)-mediated signaling have been implicated in tumor cell invasion, survival, and metastasis in a variety of cancers. This study was undertaken to investigate the biological roles of uPA and uPAR in prostate cancer cell invasion and survival, and the potential of uPA and uPAR as targets for prostate cancer therapy. uPA and uPAR expression correlates with the metastatic potential of prostate cancer cells. Thus, therapies designed to inhibit uPA and uPAR expression would be beneficial. LNCaP, DU145, and PC3 are prostate cancer cell lines with low, moderate, and high metastatic potential, respectively, as demonstrated by their capacity to invade the extracellular matrix. In this study we utilized small hairpin RNAs (shRNAs), also referred to as small interfering RNAs, to target human uPA and uPAR. These small interfering RNA constructs significantly inhibited uPA and uPAR expression at both the mRNA and protein levels in the highly metastatic prostate cancer cell line PC3. Our data demonstrated that uPA-uPAR knockdown in PC3 cells resulted in a dramatic reduction of tumor cell invasion as indicated by a Matrigel invasion assay. Furthermore, simultaneous silencing of the genes for uPA and uPAR using a single plasmid construct expressing shRNAs for both uPA and uPAR significantly reduced cell viability and ultimately resulted in the induction of apoptotic cell death. RNA interference for uPA and uPAR also abrogated uPA-uPAR signaling to downstream target molecules such as ERK1/2 and Stat 3. In addition, our results demonstrated that intratumoral injection with the plasmid construct expressing shRNAs for uPA and uPAR almost completely inhibited established tumor growth and survival in an orthotopic mouse prostate cancer model. These findings uncovered evidence of a complex signaling network operating downstream of uPA-uPAR that actively advances tumor cell invasion, proliferation, and survival of prostate cancer cells. Thus, RNA interference-directed targeting of uPA and uPAR is a convenient and novel tool for studying the biological role of the uPA-uPAR system and raises the potential of its application for prostate cancer therapy.

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mentioning

confidence: 99%

RNA Interference-directed Knockdown of Urokinase Plasminogen Activator and Urokinase Plasminogen Activator Receptor Inhibits Prostate Cancer Cell Invasion, Survival, and Tumorigenicity in Vivo

Pulukuri

Gondi

Lakka

et al. 2005

Journal of Biological Chemistry

243

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“…Moreover, it has been demonstrated that both ERK1/2 and JNK are active in Ha-Ras EJ stably transfected NIH3T3 cells, but only ERK was required for an invasive phenotype (Janulis et al, 1999). We have previously demonstrated that p38 is also required for in vitro invasion of v-Ha-Ras EJ stably transfected NIH3T3 cells (Behren et al, 2005).…”

Section: Introductionmentioning

confidence: 94%

“…It confers cellular migration and invasion, but at the same time apoptosis (Haq et al, 2002;Deng et al, 2004;Behren et al, 2005;Qi et al, 2006;Sun et al, 2007). In this study we attempted to identify downstream regulators of this pathway that specifically convey signals culminating in an in vitro invasive phenotype.…”

Section: Introductionmentioning

confidence: 99%

Phenotype-assisted transcriptome analysis identifies FOXM1 downstream from Ras–MKK3–p38 to regulate in vitro cellular invasion

et al. 2009

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The Ras oncogene is known to activate three major MAPK pathways, ERK, JNK, p38 and exert distinct cellular phenotypes, that is, apoptosis and invasion through the Ras-MKK3-p38-signaling cascade. We attempted to identify the molecular targets of this pathway that selectively govern the invasive phenotype. Stable transfection of NIH3T3 fibroblasts with MKK3 act cDNA construct revealed similar p38-dependent in vitro characteristics observed in Ha-Ras EJ -transformed NIH3T3 cells, including enhanced invasiveness and anchorage-independent growth correlating with p38 phosphorylation status. To identify the consensus downstream targets of the Ras-MKK3-p38 cascade involved in invasion, in vitro invasion assays were used to isolate highly invasive cells from both, MKK3 and Ha-Ras EJ transgenic cell lines. Subsequently a genome-wide transcriptome analysis was employed to investigate differentially regulated genes in invasive Ha-Ras EJ -and MKK3 act -transfected NIH3T3 fibroblasts. Using this phenotype-assisted approach combined with system level protein-interaction network analysis, we identified FOXM1, PLK1 and CDK1 to be differentially regulated in invasive Ha-Ras EJ -NIH3T3 and MKK3 act -NIH3T3 cells. Finally, a FOXM1 RNAknockdown approach revealed its requirement for both invasion and anchorage-independent growth of Ha-Ras EJand MKK3 act -NIH3T3 cells. Together, we identified FOXM1 as a key downstream target of Ras and MKK3-induced cellular in vitro invasion and anchorage-independent growth signaling.

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“…Additional studies also showed that p38 phosphorylation is required for K-Ras dependent invasion in pancreatic cancer (121) as well as for prostate (122) and lung cancer invasion (123). Although studies with over-expressed dominant negative p38 proteins showed that only p38alpha (but not other isoforms) is required for H-Ras-induced invasion in 3T3 cells (124), p38gamma proteins were recently shown to promote breast cancer invasion downstream of Ras (73). Furthermore, p38alpha, but not p38beta, can phosphorylate pro-invasive EGFR (epidermal growth factor receptor) at Y1045 and T669, leading to its internalization and destruction (125)(126)(127).…”

Section: The Roles Of the P38 Pathway In Regulating Cell Invasion Andmentioning

confidence: 99%

The p38 MAPK stress pathway as a tumor suppressor or more?

Loesch¹

2008

Front Biosci

100

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Abstractp38 mitogen-activated protein kinases (p38 MAPKs) are a group of serine/threonine protein kinases that together with ERK (extracellular signal-regulated kinases) and JNK (c-Jun N-terminal kinases) MAPKs act to convert different extracellular signals into specific cellular responses through interacting with and phosphorylating downstream targets. In contrast to the mitogenic ERK pathway, mammalian p38 MAPK family proteins (alpha, beta, gamma, and delta), with and without JNK participation, predominantly regulate inflammatory and stress response. Recent emerging evidence suggests that the p38 stress MAPK pathway may function as a tumor suppressor through regulating Ras-dependent and -independent proliferation, transformation, invasion and cell death by isoform-specific mechanisms. A selective activation of a stress pathway to block tumorigenesis may be a novel strategy to control human malignancies. KeywordsThe p38 MAPK pathway; Ras; Tumor Suppressor; isoform-specific; Oncogenesis; Review INTRODUCTIONMAPKs (mitogen-activated protein kinases) consist of ERK (extracellular signal-regulated kinase), JNK (c-Jun N-terminal kinase), and p38 cascades (1, 2). Classically, MAPKs function by phosphorylating substrates containing consensus sequence Ser/Thr-Pro (3) after they are phosphorylated and activated by upstream kinases (MAPK kinases). Each of these MAPK pathways has several family members but all share the same conservative Thr-XaaTyr phosphorylation motif (where Xaa is any amino-acid) (1, 2, 4). The ERK activity is mostly frequently activated by mitogens and required for cell proliferation, differentiation and/or transformation (5, 6). JNK and p38 pathways, on the other hand, are predominantly responsive to stress and cytokine signaling and play an in important role in regulating stress response and inflammation (7,8). MAPKs can be activated almost by all type of stimuli and are consequently involved in many critical biological processes such as proliferation, differentiation, cell death and transformation through regulating downstream gene expression and/or interacting with other signaling cascades.Send correspondence to: Dr. Guan Chen, Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, gchen@mcw.edu. HHS Public Access Author Manuscript Author ManuscriptAuthor Manuscript Author ManuscriptThe p38 upstream activators include MAPK kinase 6 (MKK6) and MKK3. Downstream effectors consist of kinases such as MK2 (MAPK-activating protein kinase 2) and PRAK (p38-related/activated protein kinase) as well as transcription factors including ATF-2 (activating transcription factor-2), MEF2 (myocyte enhancement factor 2), and c-Jun (4, 9). The mammalian p38 family consists of four isoform proteins (alpha, beta, gamma, and delta), with p38alpha and p38beta 75% identical in their amino-acid sequence, and p38gamma and p38delta about 60% identical to p38alpha (4, 10). p38alpha and p38beta are susceptible to inhibition by SB drugs (SB203580 and SB202190) wh...

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The p38 SAPK pathway is required for Ha-ras induced in vitro invasion of NIH3T3 cells

Cited by 16 publications

References 35 publications

RNA Interference-directed Knockdown of Urokinase Plasminogen Activator and Urokinase Plasminogen Activator Receptor Inhibits Prostate Cancer Cell Invasion, Survival, and Tumorigenicity in Vivo

RNA Interference-directed Knockdown of Urokinase Plasminogen Activator and Urokinase Plasminogen Activator Receptor Inhibits Prostate Cancer Cell Invasion, Survival, and Tumorigenicity in Vivo

Phenotype-assisted transcriptome analysis identifies FOXM1 downstream from Ras–MKK3–p38 to regulate in vitro cellular invasion

The p38 MAPK stress pathway as a tumor suppressor or more?

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