ABSTRACT
Objective@#To investigate the migration and invasion behaviors of Hep-2 after the targeted knockdown of yes-associated protein (YAP).@*Methods@#Hep-2 cells were knock-downed for YAP by shRNA as YAP-shRNA group, Hep-2 treated with non-specific shRNA as YAP-NC group, and Hep-2 with no treatment as control. Glucose uptake and lactate production in the cells were examined to assess Warburg effect. The migration and invasion behaviors of cells in three groups were observed. The expressions of vimentin and E-cadherin were detected by RT-PCR and Western Blot. The statistical software GraphPad Prism 7.0 was used to analyze significance of data. Two tailed Student′ s t-tests was used to determine significance when only two groups were compared. P values of less than 0.05 was considered statistically significant.@*Results@#Downregulation of YAP led to a obvious decrease in glucose uptake [(18.51±1.72)%] and lactate production [103.40±8.32] in Hep-2 cells compared with control [(41.20±1.11)% and 743.69±19.49, t=19.20 and 52.33, respectively, both P<0.01] and YAP-NC group [(39.60±0.78)% and 705.22±17.20, t=19.34 and 54.56, respectively, both P<0.01]. Compared with the control group (78.32±4.04) and YAP-NC group (77.28±3.11), the scratch healing ability of Hep-2 cells was significantly decreased in YAP-shRNA group (44.71±4.68). The P value was less than 0.01 (t=9.42 and 10.04). The number of cells with YAP-shRNA (33.30±4.19) passing through compartments was remarkable fewer than the control group (133.71±6.72) and YAP-NC group (126.32±4.21). The P value was less than 0.01 (t=21.96 and 27.13). The expression of E-cadherin protein in cells of YAP-shRNA group (6.16±0.11) was up-regulated compared with control (0.97±0.10, t=35.70, P<0.01) and YAP-NC group (1.13±0.09, t=36.28, P<0.01), while the expression of vimentin protein in cells of YAP-shRNA group (1.08±0.09) was down-regulated compared with control (5.67±0.12, t=29.91, P<0.01) and YAP-NC group (5.51±0.12, t=29.04, P<0.01).@*Conclusions@#The down-regulation of YAP in Hep-2 inhibits the migration and invasion of cells via suppressing Warburg and EMT program.
ABSTRACT
Objective To observe the RASSF1A expression in laryngeal keratosis and laryngeal squamous cell carcinoma and investigate the relationship between the gene and the occurrence and development of this disease. Methods The specimens of keratosis of larynx(43 cases)and laryngeal squamous cell carcinoma(31 cases)in Oto-laryngology-Head and Neck surgery of Anhui provincial hospital from Dec 2009 to Dec 2012 were collected.Nor-mal vocal fold mucosa from the cadaveric head as control group(8 cases)were collected.Immunohistochemical meth-ods were used to detect the expression of RASSF1A protein in these tissues.The diffuse distribution of brown gran-ules in the cell cytoplasm yielded positive results and the nucleus was not coloring.The percentage of positive cells of every section,and the average standard deviation(￣x±s)were calculated.SPSS17.0 statistical analysis software was used to analyze the data,P0.05).RASSF1A expression in the clinical stage from I to IV of laryngeal squamous cell carcinoma gradually de-clined.During the clinical stage,the difference had statistics significance (P0.05).ConcIusion The reduction of RASSF1A expression correlated with laryn-geal squamous cell carcinoma,which has certain significance for the different ion of carcinoma and laryngeal precan-cerous lesion.
ABSTRACT
BACKGROUND: Plenty of materials have been used as stent in total auricular reconstruction with varying advantages and disadvantages. The in vitro prepared tissue engineered complex cartilage stent has been not applied in clinic.OBJECTIVE: To observe the ultrastructure changes of surrounding tissues following total auricular reconstruction with nickel-titanium (NiTi) shape memory alloy stent in vivo.DESIGN, TIME AND SETTING: The randomized, controlled, animal experiment was performed in the Central Laboratory of EENT Hospital of Fudan University, Anhui Provincial Hospital and Department of Nuclear Medicine, Huashan Hospital of Fudan University from April 2004 to October 2005.MATERIALS: A total of 12 New Zealand rabbits were randomized divided into the skin-expanded and unexpanded groups, with 6 animals in each group.METHODS: Each animal underwent subcutaneous implantation with a NiTi alloy stent like human auricle. In the skin-expanded group, the stent was implanted after skin expansion, while no treatment in the unexpanded group.MAIN OUTCOME MEASURES: The ultrastructure changes of surrounding tissues were detected by scanning electron microscope at months 6 and 12 after implantation.RESULTS: Structure of nickel-titanium alloy implant stent make fibrovascular ingrowth more easily. The surface of nickel-titanium alloy stent was smooth without corrosion. There was no evidence of infection in the host tissue. Microstructure of Ni-Ti stent wires showed no morphologic and functional changes after subcutaneously implanted in vivo, and the difference between 2 groups had no significance.CONCLUSION: NiTi shape memory alloy stent can smoothly pass the process of fibrovascularization with less complication.According to the examination by SEM, the nickel-titanium stent showed good biocompatibility.