ABSTRACT
With the understanding of microRNA (miRNA or miR) functions in tumor initiation, progression, and metastasis, efforts are underway to develop new miRNA-based therapies. Very recently, we demonstrated effectiveness of a novel humanized bioengineered miR-124-3p prodrug in controlling spontaneous lung metastasis in mouse models. This study was to investigate the molecular and cellular mechanisms by which miR-124-3p controls tumor metastasis. Proteomics study identified a set of proteins selectively and significantly downregulated by bioengineered miR-124-3p in A549 cells, which were assembled into multiple cellular components critical for metastatic potential. Among them, plectin (PLEC) was verified as a new direct target for miR-124-3p that links cytoskeleton components and junctions. In miR-124-3p-treated lung cancer and osteosarcoma cells, protein levels of vimentin, talin 1 (TLN1), integrin beta-1 (ITGB1), IQ motif containing GTPase activating protein 1 (IQGAP1), cadherin 2 or N-cadherin (CDH2), and junctional adhesion molecule A (F11R or JAMA or JAM1) decreased, causing remodeling of cytoskeletons and disruption of cell-cell junctions. Furthermore, miR-124-3p sharply suppressed the formation of focal adhesion plaques, leading to reduced cell adhesion capacity. Additionally, efficacy and safety of biologic miR-124-3p therapy was established in an aggressive experimental metastasis mouse model
ABSTRACT
<p><b>Background</b>Endothelium allotransplantation is the primary treatment for corneal decompensation. The worldwide shortage of donor corneal tissue has led to increasing pressure to seek an alternative for surgical restoration of corneal endothelium. Compressed collagen (CC) gels have excellent biocompatibility, simple preparation course and easy to be manipulated. This study aimed to form a new biomimetic endothelium graft by CC.</p><p><b>Methods</b>We expanded bovine corneal endothelial cells (B-CECs) on laminin-coated CC to form a biomimetic endothelium graft. Scanning electron microscope was used for ultrastructural analysis and tight junction protein ZO-1 expression was tested by immunohistochemistry.</p><p><b>Results</b>The biomimetic endothelium graft, we conducted had normal cell morphology, ultrastructure and higher cell density (3612.2 ± 43.4 cells/mm). ZO-1 localization at B-CECs membrane indicated the bioengineered graft possess the basic endothelium function.</p><p><b>Conclusions</b>A.</p><p><b></b>biomimetic endothelium graft with B-CECs expanded on CC sheet was constructed, which possessed cells' morphology similar to that of in vivo endothelial cells and specific basic function of endothelium layer. This method provided the possibility of using one donor's cornea to form multiple uniformed endothelium grafts so as to overcome the shortage of cadaveric cornea tissue.</p>