Study On The Mechanism Of Cetuximab In Inhibiting The Perineural Invasion Of Salivary Adenoid Cystic Carcinoma

Objective:SACC is a malignant tumor that occurs in the salivary gland,often with PNI and distant metastasis.However,the molecular mechanism in PNI is unclear.The purpose of this experiment is to explore whether there is expression of PNI related molecules in tumor tissues of SACC patients,and how their expression compares with normal salivary gland tissues;Whether the PNI of SACC-83 can be regulated by EGFR;Can EGF and cetuximab regulate the PNI effect of SACC-83.Methods:1.We observed the expression of EGF,EGFR,MMP-2,MMP-9,Vimentin,and Ecadherin in tumor tissue and normal salivary gland tissue of SACC patients through immunohistochemical experiments.2.SACC-83 cells were treated with EGF and cetuximab together.Cell proliferation was measured by MTT,cell migration was measured by scratch test,and the perineural invasion ability of the cells was measured by Transwell test.3.The expression of PNI-related molecules in SACC-83 cells after EGF and cetuximab treatment was observed by Western blot assay.Results:1.Immunohistochemical results showed that EGF,EGFR,Vimentin,MMP-2,MMP-9 and E-cadherin were positively expressed in pathological specimens of SACCpatients.Compared with normal salivary gland tissues,EGF,EGFR,MMP-2,MMP-9 and Vimentin were highly expressed.Low expression of E-cadherin.2.MTT results showed that SACC-83 cells were treated with 50,100,150,200 and 250μg/ml cetuximab for 12,24 and 48 h,and SACC-83 cells were treated with 250μg/ml cetuximab and 160ng/ml EGF for 24 h.Cell proliferation was significantly inhibited.3.Scratch test showed that the migration ability of SACC-83 cells was significantly inhibited after being treated with 200 and 250μg/ml cetuximab and 160ng/ml EGF for 24 h.4.Transwell showed that SACC-83 cells were treated with 200 and 250μg/ml cetuximab and 160ng/ml EGF for 48 h,and their neurotropic invasion ability was significantly inhibited.5.Western Blot results showed that the protein expressions of EGFR,MMP-2,MMP-9 and Vimentin were significantly decreased and the protein expressions of E-cadherin were significantly increased after treatment of SACC-83 cells with 211μg/ml cetuximab and 160ng/ml EGF for 48 h.Conclusion:1.The expression of PNI-related molecules EGF,EGFR,Vimentin,MMP-2,MMP-9 and E-cadherin is present in SACC tumor tissue,with high expression of EGF,EGFR,MMP-2,MMP-9 and Vimentin and low expression of E-cadherin.There is a correlation between PNI and EGFR.2.EGFR can regulate the proliferation,migration and neurotropic invasion of SACC-83 cells.3.Cetuximab can competitively bind to EGFR and inhibit the binding of other ligands,decrease PNI-related molecules MMP-2,MMP-9 and Vimentin,and increase the expression of E-cadherin.4.Cetuximab can reduce the proliferation,migration and neurotropic invasion of SACC-83 cells.