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

Abstract

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.

AMPK Regulates Metabolic Actions of Glucocorticoids by Phosphorylating the Glucocorticoid Receptor through p38 MAPK

“…5A, left panel). The MAPK kinase 3 (MKK3) is known to activate p38 MAPK␣ and stimulates mRNA expression of the p38 MAPK-responsive forkhead box M1 (FOXM1) gene in 3T3-L1 cells (43). SB202190, but not PD098058, efficiently abolished the MKK3 constitutively active form (CA)-induced FOXM1 mRNA expression, suggesting that SB202190 suppressed kinase activity of the p38 MAPK␣ in these cells.…”

p38 MAPK Down-regulates Fibulin 3 Expression through Methylation of Gene Regulatory Sequences

“…It regulates different cellular functions. For example, it can inhibit proliferation (6 -8) and adhesion (9) and promote differentiation (7,10), apoptosis (11)(12)(13), migration (14), and invasion (15)(16). In addition, it can also activate proliferation (17)(18) or survival (19 -21).…”

Differential Expression of ADAM23, CDKN2A (P16), MMP14 and VIM Associated with Giant Cell Tumor of Bone

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