ABSTRACT
Progressive tumor growth is dependent on angiogenesis. The mechanisms by which endothelial cells (ECs) are incorporated to develop new blood vessels are not well understood. Recent studies reveal that the ezrin radixin moesin (ERM) family members are key regulators of cellular activities such as adhesion, morphogenetic change, and migration. We hypothesized that ezrin, one of the ERM family members, may play important roles in ECs organization during angiogenesis, and new vessels formation in preexisting tissues. To test this hypothesis, in this study, we investigated the effects of ezrin gene silencing on the migration and angiogenesis of human umbilical vein endothelial cells (HUVECs) in vitro. HUVECs were transfected with plasmids with ezrin-targeting short hairpin RNA by using the lipofectamine-2000 system. Wound assay in vitro and three-dimensional culture were used to detect the migration and angiogenesis capacity of HUVECs. The morphological changes of transfected cells were observed by confocal and phase contrast microscopy. Our results demonstrated that the decreased expression of ezrin in HUVECs significantly induced the morphogenetic changes and cytoskeletal reorganization of the transfected cells, and also reduced cell migration and angiogenesis capacity in vitro, suggesting that ezrin play an important role in the process of HUVECs migration and angiogenesis.
Subject(s)
Humans , Cell Movement , Genetics , Cytoskeletal Proteins , Genetics , Metabolism , Cytoskeleton , Metabolism , Human Umbilical Vein Endothelial Cells , Cell Biology , Metabolism , Physiology , Neovascularization, Physiologic , GeneticsABSTRACT
Progressive tumor growth is dependent on angiogenesis. The mechanisms by which endothelial cells (ECs) are incorporated to develop new blood vessels are not well understood. Recent studies reveal that the ezrin radixin moesin (ERM) family members are key regulators of cellular activities such as adhesion, morphogenetic change, and migration. We hypothesized that ezrin, one of the ERM family members, may play important roles in ECs organization during angiogenesis, and new vessels formation in preexisting tissues. To test this hypothesis, in this study, we investigated the effects of ezrin gene silencing on the migration and angiogenesis of human umbilical vein endothelial cells (HUVECs) in vitro. HUVECs were transfected with plasmids with ezrin-targeting short hairpin RNA by using the lipofectamine-2000 system. Wound assay in vitro and three-dimensional culture were used to detect the migration and angiogenesis capacity of HUVECs. The morphological changes of transfected cells were observed by confocal and phase contrast microscopy. Our results demonstrated that the decreased expression of ezrin in HUVECs significantly induced the morphogenetic changes and cytoskeletal reorganization of the transfected cells, and also reduced cell migration and angiogenesis capacity in vitro, suggesting that ezrin play an important role in the process of HUVECs migration and angiogenesis.
ABSTRACT
Objective: To study the molecular mechanism underlying androgen-independent prostate cancer invasion and metastasis promoted by vimentin. Methods: With specific amplified primer of vimentin, full-length vimentin and full-length antisense-vimentin cDNA fragments were synthesized by reverse transcriptase polymerase chain reaction ( RT-PCR). RT-PCR products were cloned into plasmid pcDNA3.1 (+) and pcDNA3.1 (-). The recombinant vimentin plasmids were transfected into androgen-independent prostate cancer PC-3M-2B4 and PC-3M-1E8 cells. The positive clones were screened by G418. The transwell chamber migration assay was used to study cell invasion. Western blotting was used to determine specific protein expression in the cells. Results: Vimentin promoted androgen-independent prostate cancer cell invasion and metastasis by regulating the E-cadherin/β-catenin complex via C-src kinase. Conclusion: Vimentin could be used as an important prognostic factor for androgen-independent prostate cancer cell invasion and metastasis.