Effects And Mechanisms Of Over-expression Of Human Cathelicidin Antimicrobial Peptide CAMP On Oral Squamous Carcinoma Cell HSC-3

Objective: Human cathelicidin antimicrobial peptide（CAMP）and its active product LL-37 have a broad spectrum of antibacterial effects.In recent years,they have also been found to play important roles in the development of various tumors.The aims of this study were to establish an oral squamous cell carcinoma HSC-3 cell lines with CAMP and C-terminal deletion of LL-37 mutant CDEL,and to study the effects and mechanisms of the expression of CAMP gene on oral squamous cell carcinoma HSC-3 cells.Methods: Using the fetal brain c DNA as a template,the open reading frame of CAMP and CDEL were cloned.The open reading frame of CAMP was cloned into the p Flag-CMV4 vector（p Flag-CAMP）,and CDEL was also cloned into the p Flag-CMV4 vector（p Flag-CDEL）.PFlag-CAMP and p Flag-CDEL eukaryotic expression vectors were transfected into HSC-3 cells,and the proteins of the obtained monoclonal cells were collected by G418,and the proteins were analyzed by Western Blot.The expression of the proteins were analyzed by immunofluorescence technique.The p Flag-CMV4 eukaryotic expression vector was transfected into HSC-3 cells,and the cells obtained after screening with G418 were used as controls.After obtaining HSC-3 cell lines stably expressing p Flag-CAMP and p Flag-CDEL,the clone formation ability and proliferation and viability were compared between p Flag-CAMP and p Flag-CDEL stably transfected HSC-3 cells and controls by colony formation assay and CCK-8 assay.Using the scratch assay and transwell invasion assay to compare the differences in migration ability and invasive ability between p Flag-CAMP and p Flag-CDEL stably transfected HSC-3 cells and controls.Western Blot analysis was used to detect apoptosis-related proteins,and the effects of p Flag-CAMP and p Flag-CDEL on the apoptosis of HSC-3 cells and their mechanisms were analyzed.Results: The open reading frame of CAMP and CDEL were cloned from the fetal brain c DNA.CAMP and CDEL were cloned into the eukaryotic expression vector p Flag-CMV4.By immunofluorescence technology and Western Blot analysis,the transfected proteins were stably expressed by transfecting HSC-3 cell lines with p Flag-CAMP and p Flag-CDEL eukaryotic expression vectors.After p Flag-CAMP and p Flag-CDEL stably transfected HSC-3 cells,the cell clonality was inhibited,and the proliferative capacity and viability of HSC-3 cells were weakened,and the migration ability and invasion ability of HSC-3 cells were also inhibited,and apoptosis was promoted.Overexpression of CAMP and CDEL in oral squamous cell carcinoma HSC-3 cells can induce Caspase-3dependent apoptosis through P53-Bcl-2/BAX signaling pathway.Conclusions:The HSC-3 cell lines stably expressing CAMP and CDEL were established.CAMP and CDEL overexpressing in oral squamous cell carcinoma HSC-3 cells can inhibit the cloning ability of HSC-3 cells and attenuate HSC-3 cells proliferation and viability,migration,invasion ability and promotion of apoptosis.