Figure 1.

Iloprost inhibits anchorage-independent growth in some but not all NSCLC. (A) The indicated NSCLC cell lines were grown in soft agar in the absence or presence of 10 µM iloprost. The colonies were incubated for several weeks and subsequently counted as described in the Materials and Methods. Data are shown as mean of three independent experiments with the SEM indicated. (B) The cell line A549 is shown to be iloprost-sensitive by soft agar, whereas H460 is seen to be an iloprost-insensitive cell line. (C) Activation of PPARγ and E-cadherin is induced by iloprost. The indicated NSCLC cell lines were transiently transfected with PPAR-RE, along with CMV-β-gal to normalize for transfection efficiency. After an overnight incubation, cells were exposed for 48 hours with 10 µM iloprost. Extracts were prepared and promoter activity was determined as luciferase units normalized to CMV-β-gal. Results represent the mean of three independent experiments with the SEM indicated. (D) The indicated NSCLC cell lines were transiently transfected with E-cadherin reporter, along with CMV-β-gal to normalize for transfection efficiency and exposed for 48 hours with 10 µM iloprost. The PPARγ antagonist compound T0070907 reduces PPAR-RE activity induced by iloprost/Fzd 9. (E) The indicated cell lines were transiently transfected with PPAR-RE, or empty vector, along with CMV-β-gal to normalize for transfection efficiency. After an overnight incubation, cells were exposed for 48 hours with 10 µM iloprost and/or 5 µM T0070907.