| Objective:To study whether the focal adhesion kinase inhibitor TAE226could inhibit the metastasis and pithelial-mesenchymal transition（EMT）progression in oral squamous cell carcinoma（OSCC）cell line HSC-3/HSC-4cells and its mechanism of action behind.we hope to provide a theoretical and experimental basis for the treatment of OSCC.Methods:1.The HSC-3 cells and HSC-4 cells were cultured with TAE226under different concentrations（0,1μM,5μM,10μM）on hour 24,48 and 72.q RT-PCR was could conducted to detect the expression of EMT related factors（TGF-β,E-cadherin,and vimentin）in these cells.2.Western Blot technique was conducted to detect the expression of EMT related proteins（TGF-β,E-cadherin,and vimentin）after TAE226 at the concentrations of 0μM,1μM,5μM,10μM was applied to HSC-3 cells and HSC-4 cells for 48 h.Results:The results of q RT-PCR showed that TAE226 promoted the expression of E-cadherin m RNA in HSC-3 cells and HSC-4 cells and inhibited the expression of TGF-βm RNA and vimentin m RNA.The expression of TGF-βm RNA and vimentin m RNA of control groups in the two cell lines increased with time while decreased in E-cadherin m RNA（P<0.05）.Although,the expression of HSC-4cell’s vimentin/TGF-βin 1μM group was not significantly different from that in control group when the time of TAE226 was 24 h（P>0.05）,vimentin/TGF-βin other experimental groups was lower than that in control group（P<0.05）and E-cadherin was higher than that in the control group（P<0.05）.At the same time,When the concentration of TAE226 increased from 1μM to 10μM,the expression of E-cadherin m RNA gradually increased,while vimentin m RNA and TGF-βm RNA decreased gradually.At the same concentration,The time of action of TAE226 extended from 24h to 48h,72h,The expression of E-cadherin m RNA also increased gradually too（P<0.05）,while TGF-βm RNA and vimentin m RNA decreased too（P<0.05）.2.After treated by TAE226 for 48 h,the expression of E-cadherin protein in HSC-3 and HSC-4 cells increased gradually in 0μM group to10μM group（P<0.05）.while The expression of TGF-βprotein and vimentin protein decreased gradually（P<0.05）.It meant that TAE226 promoted the expression of E-cadherin protein and inhibited the expression of TGF-βand vimentin protein in two cell lines,and the effection was more obvious with the increase of TAE226 dose.3.Correlation between TGF-βand E-cadherin,vimentin in HSC-3 cells and HSC-4 cells results.Pearson correlation analysis showed that the relative expression of TGF-βwas negatively Correlated with E-cadherin in two cell lines(r HSC-3=-0.795,p HSC-3=0.000;r HSC-4=-0.714,p HSC-4=0.000),but was positively correlated with vimentin(r HSC-3=0.922,p HSC-3=0.000;r HSC-4=0.921,p HSC-4=0.000)Conclusions:1.TAE226 promoted the expression of E-cadherin in HSC-3and HSC-4 cells,while inhibited the expression of vimentin in two cell lines.The effect was gradually enhanced With the increase of TAE226 dosage.2.TAE226 inhibited EMT process of HSC-3 cells and HSC-4 cells,It indicated that TAE226 inhibited the metastasis of oral squamous carcinoma cells.3.TGF-βm RNA and protein in oral squamous cell line HSC-3 and HSC-4 cells can be effectively inhibited by TAE226,and the inhibitory effect increased with the increased of TAE226 dosage.The changes of vimentin,TGF-βand E-cadherinine were correlated with each other,which suggested that TAE226may inhibit the progression of EMT by down-regulating TGF-β. |
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