Figure 5

Elevated eIF4E expression controls the synthesis of Pim-1 in NSCLC. H1299 and A549 cells were transfected with eIF4E siRNA at a final concentration of 50nM. EIF4E and Pim-1 mRNAs were detected in H1299 (A) and A549 cells (B) using qRT-PCR 48 h posttransfection. Pim-1 and eIF4E protein expression was detected in H1299 (C) and A549 cells (D) using Western Blot assay 48 h psottransfection. (E) Cell growth was evaluated in A549 and H1299 cells after transfection with si-eIF4E for 0-96 h using MTT assay. (F) The ability of cell motility was evaluated in A549 and H1299 cells after transfection with si-eIF4E for 48 h using wound scratch assay. A uniform scratch was made in each confluent layer culture, the extent of wound closure was monitored under microscope, and photographs were taken at 0 and 48 h. Data are shown as mean ± SD. Similar results were obtained in three independent experiments. *P < 0.05. (G) EIF4E expression is frequently elevated in squamous cell carcinoma (a: SP, 100×; b: a segment magnified at 200×) and adenocarcinoma of lung by immunohistochemistry analysis (c: SP, 100×; d: a segment magnified at 200×).