CH25H affects laryngeal squamous carcinoma progression through regulating autophagy and apoptosis
Objective To investigate the effect of cholesterol 25-hydroxylase(CH25H)on the progression of laryngeal squamous cell carcinoma(LSCC)and its relation with autophagy and apoptosis of tumor cells.Methods Human LSCC AMC-HN-8 cells were tansfected with CH25-plasmid or blank plasmid to construct AMC-HN-8/CH25H cell line and transfection control cell line AMC-HN-8/Mock,and untransfected AMC-HN-8 cells served as blank control.The expression level of CH25H mRNA was detected by real-time fluorescence quantitative PCR.The expression levels of CH25H protein,apoptosis-related proteins(Bcl-2,Bax and C-caspase-3),autophagy-related proteins(LC3 Ⅱ,Beclin-1 and p62),and PI3K/Akt pathway proteins(p-PI3K,PI3K,p-Akt,and Akt)were detected by Western blot.Flow cytometry was used to detect the cell apoptosis rate,and immunofluorescence was used to detect the expression of LC3 Ⅱ.The nude mice were transplanted with AMC-HN-8,AMC-HN-8/Mock,and AMC-HN-8/CH25H cells(n=8 in each group),respectively.The tumor volume was measured,and the apoptosis protein and autophagy protein levels were detected in each group.Results Compared with AMC-HN-8 and AMC-HN-8/Mock cells,the expression of CH25H in AMC-HN-8/CH25H cells was upregulated(P<0.05),the apoptosis rate was significantly increased(P<0.05);the expression of Bax and C-caspase-3 was enhanced,and Bcl-2 expression was inhibited.Meanwhile,the expression of LC3 Ⅱ and Beclin-1 was increased,and the expression of p62 was inhibited,indicating that overexpression of CH25H promoted apoptosis and autophagy in laryngeal squamous cell carcinoma cells.Furthermore,CH25H inhibited the activation of the PI3K/Akt oncogenic signaling pathway.Overexpression of CH25H inhibited LSCC tumor growth in xenografted nude mice.Conclusion CH25H regulates autophagy and apoptosis through the PI3K/Akt signaling pathway in laryngeal squamous cell carcinoma cells,thereby affecting tumor progression.
Cholesterol 25-hydroxylaseLaryngeal squamous cell carcinomaAutophagyApoptosisPhospha-tidylinositol 3-kinase/protein kinase B