Sp1Mediates MiR-375-regulated Growth And Invasion Of Head And Neck Squamous Cell Carcinoma

Objective:Recent research of proteomics found that Sp1is a key transcription factors in the metastasis of head and neck squamous cell carcinoma (HNSCC). Sp1has been found significantly upregulated in multiple types of cancer. However, the underlying mechanisms of Sp1in tumorigenesis remain unknown. One aim of our study is to investigate the affection of Spl in the proliferation and invasion of HNSCC cell. MicroRNAs (miRNAs) are evolutionarily conserved, small noncoding RNAs that are believed to play an important role in tumorigenesis. Another aim of our study is to investigate the affection of miRNA in the Spl expression level and proliferation and invasion of HNSCC cell.Methods:①The Sp1siRNA was employed to downregulate Sp1expression in HNSCC cell line Tu686. Then, CCK-8was conducted to assess cell proliferation and transwell chambers test was conducted to assess cell invasion.②Bioinformatics software was used to predict the miRNAs that target Sp1.③Western blot was conducted to examine the expression of Spl while miR-375was up regulated.④The miR-375mimic was employed to upregulate miR-375expression in HNSCC cell line Tu686. Then CCK-8was conducted to assess cell proliferation and transwell chambers test was conducted to assess cell invasion.⑤The Spl cDNA was employed to upregulate Spl of miR-375-upregulated Tu686cells. Then transwell chambers test was used to assess cell invasion.Results:①Proliferation assay:the growth of Tu686cell is inhibited while silence the expression of Sp1, and the inhibition is obvious in48h (P<0.05) and72h (P<0.01). Invasion assay:the invasion of Tu686cells were greatly inhibited compared with the empty control and negative control cells (65±15%vs126±7%and114±20%, P<0.05).②Western blot analysis showed that Sp1expression was downregulated while miR-375was up regulated.③Proliferation assay:the growth of Tu686cell is inhibited while upregulated the expression of miR-375, and the inhibition is obvious in96h (P<0.05). Invasion assay:the invasion of Tu686cells were greatly inhibited compared with the empty control and negative control cells (96±19%vs151±18%and165±21%, P<0.05).④Invasion assay:the invasion of miR-375-Sp1-upregulated Tu686cells were greatly enhanced compared with the control cells (96±17%vs56±16%,P<0.05).Conclusion:Spl mediates miR-375-regulated growth and invasion of head and neck squamous cell carcinoma.