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. Author manuscript; available in PMC: 2006 May 28.
Published in final edited form as: J Biol Chem. 2002 Oct 21;277(52):50828–50833. doi: 10.1074/jbc.M210707200

Fig. 1.

Fig. 1

A, PGE2 augments CD44 expression in NSCLC. A549 cells (106 cells/ml) were cultured in complete medium. Following overnight incubation, the cells were treated with dmPGE2 (2.5, 5.0, and 10.0 μg/ml) or vehicle (control) for 4 h. Compared with control, a 3-fold increase in CD44 expression is seen at 5.0 (*, p < 0.01) and 10.0 μg/ml dmPGE2 (*, p < 0.01). MFI, mean fluorescence intensity. The data represent the mean ± S.D. of triplicate determinations in one representative experiment of three separate experiments. B, anti-PGE2 inhibits IL-1β induced CD44 expression. A549 cells (106 cells/ml) were cultured in the presence or absence of IL-1β (280 units/ml) for 24 h. For antibody treatment, the cells were incubated with control or anti-PGE2 antibody prior to the addition of IL-1β. Compared with the control, a 2.5-fold increase in CD44 expression is seen in IL-1β treated cells (p < 0.05). Anti-PGE2 significantly reduced the capacity of IL-1β to induce CD44 (*, p < 0.05). The data represent the mean ± S.D. of triplicate determinations in one representative experiment of three separate experiments. C, agents that impact G-coupled protein receptor signaling and cAMP level modulate CD44 expression. A549 cells (106 cells/ml) were cultured in complete medium with various agents for 24 h. CD44 expression was analyzed by flow cytometry. Anti-PGE2 significantly decreased constitutive CD44 expression. Agents that increase intracellular cAMP levels, including dmPGE2 (10 μg/ml), forskolin (100 μm), IBMX (100 μm), dibutyryl cAMP (100 μm), and cholera toxin (CTX; 1 μg/ml) were potent inducers of CD44 expression. Dibutyryl cGMP (200 μm) served as a negative control and did not induce CD44 expression. Pertussis toxin (PTX; 200 ng/ml) treatment significantly decreased constitutive as well as PGE2-induced CD44 expression. Asterisks denote a statistically significant increase or decrease compared with control values (*, p < 0.01; **, p < 0.05). The data represent the mean ± S.D. of triplicate determinations in one representative experiment of three separate experiments.