The aim of this study is to characterize the phenotype ofpancreatic ductal adenocarcinoma (PDAC) cells in relationto the expression of epithelial-to-mesenchymal transition(EMT) markers and determine whether ukrain, ananticancer drug based on the alkaloids extracted fromgreater celandine, modulates in vitro the malignantbehavior of PDAC cells in order to extend ourunderstanding of its therapeutic potential. Three cell lines(HPAF-II, HPAC, and PL45) were treated with ukrain (5, 10,and 20 lmol/l) for 48 h or left untreated (control). Cellproliferation was assessed by growth curves. Apoptosiswas determined by Hoechst nuclear staining and bycytochrome c and caspase-8 expressions. The EMTmarkers E-cadherin, b-catenin, and vimentin, as well asactin and tubulin cytoskeletons, were analyzed byimmunofluorescence. Interphase and mitotic microtubulesas well as abnormal mitotic figures were studied byfluorescence microscopy after tubulin immunolabeling.Ukrain strongly suppressed cell proliferation and inducedapoptosis possibly through an extrinsic pathway ascytochrome c immunoreactivity suggested that the integrityof the mitochondria was not affected. Tubulin expressionindicated an antiproliferative effect of ukrain on the basis ofalterations in mitotic spindle microtubule dynamics,leading to abnormal mitosis. Membranous E-cadherin/b-catenin immunoreactivity was similarly expressed incontrol-treated and ukrain-treated cells, although the drugupregulated E-cadherin in cell lysates. Our results suggestthat ukrain exerts its chemotherapeutic action on PDACcells targeting mitotic spindle microtubules, leading toabnormal mitosis and apoptosis, and favoring cellcohesiveness. The differentiated epithelial phenotype ofHPAF-II, HPAC, and PL45 cell lines concomitant with ahighly invasive potential suggests that further experimentswill be necessary to definitively clarify the role of EMT inPDAC progression.
|Titolo:||Pancreatic cancer cells retain the epithelial-related phenotype and modify mitotic spindle microtubules after the administration of ukrain in vitro|
|Data di pubblicazione:||2012|
|Appare nelle tipologie:||1.1 Articolo in rivista|