The Role Of CXCL12 And CXCR4 In Perineural Invasion Of Salivary Adenoid Cystic Carcinoma

Objective:1.To detect the expression of the chemokine CXCL12 and its receptor CXCR4 in human schwann cells(HSC) and salivary adenoid cystic carcinoma cells(ACC-3).2.To observe the chemotaxis by the supernatant of Human Schwann cells in ACC-3 cells,which might be inhibited by AMD3100,which is activation antagonist of chemokine receptor CXCR4.In order to study the role of the biological axis of CXCL12/CXCR4 in the perineural invasion of salivary adenoid cystic carcinoma.Methods:There were two sections in this experiment:1.Quantitative real-time PCR was applied to detect the expression of the chemokine CXCL12 and its receptor CXCR4 in mRNA level in human schwann cells and salivary adenoid cystic carcinoma cells.lmmunofluorescence was applied to detect the positive protein of the chemokine CXCL12 in human schwann cells and the positive protein of the chemokine receptor CXCR4 in salivary adenoid cystic carcinoma cells.Flow cytometry was applied to detect the expression of the chemokine receptor CXCR4 on membrans of human schwann cells and salivary adenoid cystic carcinoma cells.Western blot was applied to detect the chemokine CXCL12 in the supernatant of human schwann cells and salivary adenoid cystic carcinoma cells.ELISA was applied to observe the CXCL12 secretion induced in human schwann cells after being cultured with methotrexate.2.Chemotaxis assay was applied to observe the the chemotaxis by supernatant of human schwann cells in ACC-3 cells,which might be inhibited by AMD3100,which is activation antagonist of chemokine receptor CXCR4.All the data of the experiment was analyzed by the SPSS 10.0 statistical software,Student's test.Results:1.The expressions of chemokine CXCL12 were high in mRNA level in human schwann cells,but very low in ACC-3 cells.On the other side,the chemokine receptor CXCR4 were both expressed in mRNA level in human schwann cells and ACC-3 cells.The positive protein expressions of chemokine CXCL12 were high in human schwann cells,the positive protein expressions of chemokine receptor CXCR4 were high in ACC-3 cells.The positive rate of CXCR4 expression was significantly higher in HSC(67.10%) and ACC-3(74.15%).The chemokine CXCL12 was found in the supernatant of human schwann cells,but it was not found in ACC-3 cells'. Methotrexate inhibited the CXCL12 secretion induced in Human Schwann cells,with statistical significance.2.The chemotaxis by the supernatant of human schwann cells in ACC-3 cells was observed,which could be inhibited by AMD3100,which is activation antagonist of chemokine receptor CXCR4.Conclusions:1.The chemokine CXCL12 could be expressed and excreted by human schwann cells.The protein of the chemokine receptor CXCR4 was expressed on the membrane of human schwann cells.The chemokine receptor CXCR4 could be expressed by ACC-3 cells.ACC-3 cells seldom expressed the chemokine CXCL12.The methotrexate can significantly inhibited the CXCL12 secretion induced in Human Schwann cells.2..The chemotaxis of the supernatant of human schwann cells in ACC-3 cells may be mediated by the chemokine CXCL12 and its receptor CXCR4.It can be significantly inhibited by AMD3100.