FGF8 induces epithelial-mesenchymal transition and promotes metastasis in oral squamous cell carcinoma / 国际口腔科学杂志·英文版

Oral squamous cell carcinoma (OSCC) is one of the most common cancers worldwide, and with 354 864 new cases each year. Cancer metastasis, recurrence, and drug resistance are the main causes to cripples and deaths of OSCC patients. As potent growth factors, fibroblast growth factors (FGFs) are frequently susceptible to being hijacked by cancer cells. In this study, we show that FGF8 is upregulated in OSCC tissues and high FGF8 expression is related with a set of clinicopathologic parameters, including age, drinking, and survival time. FGF8 treatment enhances the invasive capability of OSCC cells. Lentivirus-based FGF8 expression promotes OSCC metastasis in a mouse lung metastasis model. Further, mechanistic study demonstrates that FGF8 induces epithelial-mesenchymal transition (EMT) in OSCC cells. These results highlight a pro-metastatic function of FGF8, and underscore the role of FGF8 in OSCC development.

Hypospadias induced by maternal exposure to di-n-butyl phthalate and its mechanisms in male rat offspring / 中华男科学杂志

Objective@#To induce hypospadias in male rat offspring by maternal exposure to di-n-butyl phthalate (DBP) during late pregnancy and further investigate its mechanisms.@*METHODS@#We randomly divided 20 pregnant rats into a DBP exposure and a control group, the former treated intragastrically with DBP while the latter with soybean oil at 750 mg per kilogram of the body weight per day from gestation days (GD) 14 to 18. On postnatal day (PND) 1, we recorded the incidence rate of hypospadias and observed the histopathological changes in the genital tubercle of the hypospadiac rats. We also measured the level of serum testosterone (T) by radioimmunoassay and determined the mRNA and protein expressions of the androgen receptor (AR), sonic hedgehog (Shh), bone morphogenetic protein 4 (Bmp4) and fibroblast growth factor 8 (Fgf8) in the genital tubercle by real-time PCR and Western blot.@*RESULTS@#No hypospadiac male rats were found in the control group. The incidence rate of hypospadias in male offspring was 43.6% in the DBP-treatment group. Histological analysis confirmed hypospadiac malformation. The serum testosterone concentration was decreased in the hypospadiac male rats as compared with the controls (［0.49 ± 0.05］ vs ［1.12 ± 0.05］ ng/ml, P <0.05). The mRNA expressions of AR, Shh, Bmp4 and Fgf8 in the genital tubercle were significantly lower in the hypospadiac male rats than in the controls (AR: 0.50 ± 0.05 vs 1.00 ± 0.12, P <0.05; Shh: 0.65 ± 0.07 vs 1.00 ± 0.15, P <0.05; Bmp4: 0.42 ± 0.05 vs 1.00 ± 0.13, P <0.05; Fgf8: 0.46 ± 0.04 vs 1.00 ± 0.12, P <0.05), and so were their protein expressions (AR: 0.34 ± 0.05 vs 1.00 ± 0.09, P <0.05; Shh: 0.51 ± 0.07 vs 1.00 ± 0.12, P <0.05; Bmp4: 0.43 ± 0.05 vs 1.00 ± 0.11, P <0.05; Fgf8: 0.57 ± 0.04 vs 1.00 ± 0.13, P <0.05).@*CONCLUSIONS@#Maternal exposure to DBP during late pregnancy can induce hypospadias in the male rat offspring. DBP affects the development of the genital tubercle by reducing the serum T concentration and expressions of AR, Shh, Bmp4 and Fgf8 in the genital tubercle, which might underlie the mechanism of DBP inducing hypospadias.

The similarity between human embryonic stem cell-derived epithelial cells and ameloblast-lineage cells / 国际口腔科学杂志·英文版

This study aimed to compare epithelial cells derived from human embryonic stem cells (hESCs) to human ameloblast-lineage cells (ALCs), as a way to determine their potential use as a cell source for ameloblast regeneration. Induced by various concentrations of bone morphogenetic protein 4 (BMP4), retinoic acid (RA) and lithium chloride (LiCl) for 7 days, hESCs adopted cobble-stone epithelial phenotype (hESC-derived epithelial cells (ES-ECs)) and expressed cytokeratin 14. Compared with ALCs and oral epithelial cells (OE), ES-ECs expressed amelogenesis-associated genes similar to ALCs. ES-ECs were compared with human fetal skin epithelium, human fetal oral buccal mucosal epithelial cells and human ALCs for their expression pattern of cytokeratins as well. ALCs had relatively high expression levels of cytokeratin 76, which was also found to be upregulated in ES-ECs. Based on the present study, with the similarity of gene expression with ALCs, ES-ECs are a promising potential cell source for regeneration, which are not available in erupted human teeth for regeneration of enamel.

Molecular mechanism of FGF8b regulation of epithelial-mesenchymal transition in prostate cancer cells / 中南大学学报(医学版)

OBJECTIVE@#To explore the molecular mechanism of fibroblast growth factor 8b (FGF8b) in promoting epithelial-mesenchymal transition in prostate cancer DU145 cells.@*METHODS@#Cells were selected in three groups as follows: a block control group (DU145 cells), a negative control group [DU145 cells transfected with empty plasmid (pcDNA3.1/DU145)], and an experimental group [DU145 cells transfected with FGF8b (FGF8b/DU145)]. The activity of extracellular regulated protein kinases1/2( ERK1/2) pathway was detected by western-blot in the three groups. The FGF8b-DU145 cells and DU145 cells were cultured with PD98059 (an ERK kinase inhibitor) to observe microscopically the morphology changes within the cells. The experimental samples were also divided into four groups: FGF8b/DU145 cells cultured with 2% FBS (Group A); FGF8b/DU145 cells cultured with 2% FBS+PD98059 (50 μmol/L) (Group B); DU145 cells cultured with 2% FBS (Group C); DU145 cells cultured with FBS+PD98059 (50 μmol/L) (Group D). The expression of epithelial- mesenchymal transition (EMT) markers (E-cadherin, vimentin) were detected by western-blot analysis and the cell's mobility were detected by the Transwell chamber.@*RESULTS@#The activity of ERK1/2 in the experimental group was significantly higher than that in the other two control groups; when ERK kinase inhibitor PD98059 was added to FGF8b/ DU145 cells, the expression of epithelial marker E-cadherin protein was significantly increased in group B compared with that in the group A (P<0.05). The expression of mesenchymal marker vimentin protein was significantly reduced in group B compared with that in group A (P<0.05). The cell migration assay suggested that cell migration was markedly decreased in group B (P<0.05) compared with that in group A.@*CONCLUSION@#EMT in prostate cancer induced by FGF8b can be mediated by ERK kinase pathway, in which mitogen-activated/extraceluer signal regulated kinase 1 (MEK1) may be a key factor. MEK1 could be an effective target in regulating the invasion and migration of prostate cancer.

[ effects of FGF8 and Shh on expression of dopaminergic markers from human amniotic epithelial cells]

Recently, interest has increased in the potential employment of embryonic stem cells for the treatment of Parkinson's disease, which has been considered as an alternative therapeutic strategy. Due to their pluripotent differentiation potential, the finding that they do not induce carcinoma and the fact that they do not raise the ethical concerns connected with human embryonic stem cells, human amniotic epithelial cells are considered to be a very promising cell source. The aim of this study was to investigate the effects of FGF8 and Shh on the expression of dopaminergic markers from human amniotic epithelial cells. In this study, we examined the differentiation of dopaminergic neurons in vitro from AECs using the expression of several markers including TH, DAT and D beta H. For dopaminergic differentiation, sonic hedgehog [Shh] and FGF8 were added to cultures and the cultures were allowed to differentiate for 21 days. Analysis of AECs derived dopaminergic neurons was performed at the TH, DAT, beta-tubulin III and D beta H expression levels by immunocitochemistry. The significance of the data was tested by Student's t-test [between two groups] and one-way analysis of variance [ANOVA] followed by Tukey post-test. [p<0/01, p<0/05]. Combination of Shh and FGF8 showed the higher level of TH in comparison to control group or these factors alone. Moreover, Shh is more effective than FGF8 on DAT expression in comparison to expression of D beta H. These results show the capability of AECs to express dopaminergic neural markers and this ability is affected by Shh and FGF8

In vitro study on cranial neural crest differentiating into ectomesenchymal cell of the first branchial arch by FGF-8 / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of FGF-8 on cranial neural crest cell (CNCC) differentiating into ectomesenchymal cell of the first branchial arch, and determine the appropriate dose and stage of CNCC exposure to FGF-8.</p><p><b>METHODS</b>Cranial neural crest explants were cultured in free-serum medium containing modified DMEM/F12 and different doses of FGF-8. The differentiation type of CNCC were determined by in situ hybridization for Hoxa2 and immunocytochemistry for vimentin.</p><p><b>RESULTS</b>Pre-emigrating CNCC demonstrated the negative Hoxa2 stain and positive vimentin stain after treated by 100 ug/FGF-8. Both post-emigrating CNCC group and control group were positive for Hoxa2 and vimentin stain.</p><p><b>CONCLUSIONS</b>On the early stage of CNCC emigration, the first branchial arch phenotype of CNCC could be induced by FGF-8. This experiment could provide in vitro model for study on the mechanism of tooth-jaw regeneration.</p>