Effects Of NRP-1on Mediating The Differentiation Of Human Tongue Squamous Cell Carcinoma

Objective: It has been considered that the expression of NRP-1in tongue squamouscell carcinoma tissue is much higher than that in normal tongue tissue in our previousstudies. To find the important role that NRP-1mediates in cell differentiation oftongue squamous cell carcinoma, we investigated the relationship between the levelof expression of NRP-1and histopathological differentiation in tongue squamous cellcarcinoma tissue samples, and down-regulatting the expression of NRP-1in SCC25.Methods:1. The expression of NRP-1was tested via immunohistochemistry in40tonguesquamous cell carcinoma samples. Then the relationship between the level ofexpression of NRP-1and histopathological differentiation in tongue squamous cellcarcinoma tissue samples was analysed.2. Lentiviral plasmid containing siRNAs targeting NRP-1was constructed toknockdown NRP-1in SCC25cells. Real-time RT-PCR and western blotting wereemployed to detect the expression of NRP-1after knockdown of NRP-1.3. The migratory ability of SCC25cells after knockdown of NRP-1was detachedvia scratch assay and transwell comparing to the control group, and the expression ofepithelial markers E-cadherin, mesenchymal markers fibronectin and vimentin wastested employing real-time RT-PCR and western blotting.Results:1. The level of expression of NRP-1significantly correlated with thehistopathological differentiation, high level of expression of NRP-1in tonguesquamous cell carcinoma corresponding with poor histopathological differentiation. 2. Lentiviral plasmid containing siRNAs targeting NRP-1was constructedsuccessfully. Real-time RT-PCR and western blotting showed that the expression ofNRP-1in SCC25cells sharply decreased to35%.3. Scratch test and transwell showed that the cell migration was reducted afterknockdown of NRP-1via short hairpin RNA, Similarly, western blotting andreal-time RT-PCR showed that knockdown of NRP-1increased E-cadherin anddecreased fibronectin and vimentin expression in SCC cells, indicating a maturedifferentiated phenotype.Conclusions: The expression of NRP-1relates to the histopathological differentiationin tongue squamous cell carcinoma. NRP-1plays an important role in mediatingdifferentiation of SCC25, the mechanism how it works may through increasingE-cadherin and decreasing fibronectin and vimentin. The migraton of SCC25decreases after knockdown of NRP-1.