Alterations in mRNA and protein profiles in eIF4E-transfected human lung carcinoma cells

摘要: Previous work has shown that treatment of the lung cell carcinoma line DLKP with differentiation modulating agent bromodeoxyuridine (BrdU) causes posttranscriptionally regulated changes in expression growth and related genes. These gene were found to coincide an increase level phosphorylation translation initiation factor eIF-4E. In this study we have overexpressed eIF4E cells determine its role mediating seen BrdU what effects it may on cancer general as studies play a regulating carcinogenesis We also analysed overexpression Ser209 mutated non-phosphorylatable (S209) site translational functional cells. The exact is currently unknown conflicting views emerged whether necessary for regulation by eIF4E. Stable transfections carried out using wild type (4E), S209 mutant (4E-S209) HA (hemagluttinm) epitope-tagged human constructs Stable earned empty pcDNA plasmid vector negative control .Western blot analysis showed transfected HA-tagged eEF4E protein was effectively cloned limiting dilution clones chosen further analysis. Two expressing tagged eIF4E, two 4E-S209 been study. An eDF4E overexpressing clone which expresses high increased keratin 8 expression. pi integrm not other indicating levels 4E induce Immunocytochemical alpha subunits 3 cells. Invasion assays performed detected certain cancers. displayed large invasiveness compared whereas did display mvasiveness eIF4E-S209 similar controls. Large scale undertaken novel 2D-DIGE (2 dimensional-differential gel analysis) dimensional electrophoresis technique Differentially expressed proteins identified mass-spectrometry based techniques. Among involved mRNA processing, degradation cytoskeletai Of particular interest, number dynamics whose down A common regulatory element these led development hypothesis localised proteins. possible involvement represents aspect contribute oncogenesis. Changes processing indicate post-transcriptional processes apart from important Oligonucleotide microarray genes conducted transcriptional downstream regulation. Microarray there are diverse cellular functions. This indicates occur result be altered cells. Microarray signalling FAK (focal adhesion kinase) downregulated both overexpesssing Western at Immunofluorescent staining localisation affect cells. A proteomic anlysis actin dynamics. Cells therefore structures. Major structures protein. We depth into eIF4E.