CRF2 signaling alters cell adhesion in favor to cell migration and invasion.

<p>(A) HT-29 were treated or not with Ucn3 (100 nM) 30 min before adhesion and then plated on either COIV or LN-332. Cell-ECM adhesion was evaluated at 60 min with the use of a Cell Titer Aqueous MTT reagent kit. Values are normalized to the adhesion without Ucn3. Data represent the mean +/− SEN of 4 separate experiments. Effect of Ucn3 on HT-29 cells transmigration after 72 hrs was determined using a transwell assay (B) and on HT-29 cell invasion after 48 hrs by a matrigel assay (C). In both assays cells were visualized by phase contrast microscopy and counted. A phase contrast image has been joined. A circle has indicated some migrating cells and some invaded cells have been indicated by a white arrow. Data (n>3) are given as mean +/− SEM. (D) Effects of Ucn3 on HT-29 cell viability. (E) Regulation of MMP3 and MMP7 mRNA expression analyzed by RT-PCR in HT-29 cells treated with Ucn3 100 nM. (F) Immunoblot analysis of MMP3 and MMP7 protein expression in HT-29 cells following the time course of Ucn3 treatment. (G) Zymogram of gelatinolytic activities in supernatant of HT-29 cells treated or not with Ucn3 (100 nM) during 24 and 48 hrs.