down cells formed larger aggregates compared to control cells, indicating that activin A controls of cell-cell contacts via regulation of E-cadherin. Next, the effects of activin A on adhesive properties of the cells were investigated on surfaces coated with type I collagen, fibronectin and laminin. In general, activin A significantly increased the adhesion of HaCat cells on surfaces coated with type I collagen, fibronectin and laminin, whereas follistatin reduced adhesion significantly to surfaces coated with type I and fibronectin. Adhesion of shINHBA cells was significantly higher than 11 / 22 Activin A Overexpression in Oral Cancer Fig 3. Survival curves based on immunoexpression of activin A. Overall survival, as determined by the period between the treatment beginning until death or last follow-up information, and disease-free survival, time between initiation of treatment until diagnosis of the recurrence or last follow up information for those without recurrence. Censored deaths are indicated by vertical lines. The univariate analysis revealed that high positivity for activin A is significantly associated with shortened overall survival. doi:10.1371/journal.pone.0136599.g003 shControl cells in all 3 coated-surfaces. Activin A significantly augmented migration of HaCat cells, whereas follistatin significantly inhibited the migration and invasiveness of SCC-9 ZsGreen LN-1 parental cells. shINHBA cells showed significantly lower migration and invasion compared with control cells. The number of filopodia and lamellipodia in shINHBA cells was also significantly lower compared with shControl cells. Downregulation of miR143/miR145 cluster is associated with INHBA overexpression in OSCC Since previous study showed that 
miR-145 regulates features similar to those regulated by activin A in OSCCs, and to gain insight into the molecular mechanism by which activin A is overexpressed in OSCCs, we determined the relationship between INHBA mRNA SKI-II levels and expression of miR-143 and miR-145 in a series of OSCC cell lines and fresh tumor samples. For both microRNAs, a significant and inverse correlation with INHBA levels was observed in the cell lines and PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19729663 fresh tumor samples. To determine whether miR-143 and miR-145 regulate INHBA mRNA, SCC-9 and SCC-9 ZsGreen LN-1 cells were transfected with miR-143 and miR-145 mimics. Fig 10A shows the Human 3D skin equivalents are in vitro PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19728767 models used to conduct experiments on processes involving the skin e.g. disease progression and drug discovery. They are widely used for several reasons, but mainly because many studies report differences in phenotype, cellular signaling, cell migration, and drug responses when the same cells are grown under 2D vs 3D conditions. When lifted to the air interface HSEs differentiate and stratify, recapitulating the complex and dynamic environment found in human skin, meaning that data generated using these models is often fairly representative of an in vivo response. Currently however, the utility 1 / 16 Pigmented Induced Pluripotent Stem Cell-Derived Skin Models of HSEs is limited since these models lack the full cellular complexity of human skin i.e. they contain few cell types and no appendages. One major reason for this is that these models consist of cells isolated from freshly discarded tissue following surgery. This tissue is often small and therefore it is usually only possible to isolate and expand in culture the most numerous cell types i.e. dermal fibroblasts and epi
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