| Salivary adenoid cystic carcinoma is a head and neck malignancy with a low incidence.Neural infiltrative growth is one of its characteristic biological behaviors.This feature has brought many problems to clinical treatment of adenoid cystic carcinoma.Therefore,it is of great significance to explore the biological markers and mechanisms of neural invasion in salivary adenoid cystic carcinoma.At present,the relationship between chemokines and their receptors and the invasion and metastasis of malignant tumors has increasingly become a research hotspot,but the correlation between chemokine CCL20 and its receptor CCR6 and nerve invasion of salivary adenoid cystic carcinoma has not been seen at home and abroad.Objective:1.To investigate the role of CCR6 in pathological typing of salivary gland adenoid cystic carcinoma and its role in the process of neural invasion of salivary adenoid cystic carcinoma;2.Detecting the expression of CCR6 in salivary gland adenoid cystic carcinoma cells SACC-LM,and in vitro experiments to explore the CCL20/CCR6 biological axis in salivary adenoid cystic carcinoma cells SACC-LM during proliferation,metastasis and invasion effect.Method:1.The expression of CCR6 in salivary adenoid cystic carcinoma and normal salivary gland specimens was detected by immunohistochemical technique.The expression of CCR6 in both specimens and the different pathological features of salivary adenoid cystic carcinoma were analyzed and analyzed.2.The expression of chemokine receptor CCR6 in salivary adenoid cystic carcinoma cells SACC-LM was verified by immunocytochemistry and immunofluorescence.3.Stably transfected salivary adenoid cystic carcinoma cell lines that interfere with CCR6 expression by RNA interference.4.The QPCR technique and immunofluorescence technique were used to detect the inhibition efficiency of CCR6 gene and protein expression in stably transfected cell lines that inhibited the expression of chemokine receptor CCR6.5.Using CCK-8 proliferation experiment to observe the effect of CCR6-siRNA on the proliferation of salivary adenoid cystic carcinoma cell line SACC-LM.The effects of CCL20/CCR6 biological axis on chemotaxis,invasion and movement of salivary adenoid cystic carcinoma cells were explored by chemotaxis,invasion and scratch experiments.Result:1.The chemokine receptor CCR6 was positive in 22 cases of adenoid cystic carcinoma of the salivary gland.The positive expression rate was 73.33%.The positive expression of CCR6 has nothing to do with the pathological type of salivary adenoid cystic carcinoma.It has a certain correlation with its neural infiltration phenomenon,and it is not expressed in all 22 normal human salivary gland specimens.2.In the results of immunofluorescence staining and immunocytochemistry staining,salivary gland adenoid cystic carcinoma cells were significantly expressed CCR6,suggesting that this cell line can be used to study the CCL20/CCR6 biological axis and salivary adenoid cystic carcinoma peripheral nerve invasion Relevance research.3.In the CCK-8 proliferation experiment,on the second day after inoculation,the proliferation rate of CCR6-siRNA stably transfected cells was significantly lower than that of the control group.4.In the scratch test,the scratch repair effect was observed at 24 h and 48 h,and the cell viability of CCR6-siRNA stably transfected cells was significantly lower than that of the control group.5.In in vitro invasion experiments,the ability of cells that interfere with the expression of chemokine receptor CCR6 to pass through the artificial basement membrane was significantly lower than that of the control group.In chemotaxis experiments,the chemokine CCL20 was within a certain concentration range(10-200 ng/ml)can promote the motility of the cells,but has no apparent effect on cells that interfere with CCR6 expression.Conclusion:1.In the immunohistochemical specific staining results,no expression of CCR6 was found in normal salivary gland tissues,but the expression rate was 73.33% in adenoid cystic carcinoma.The results were statistically analyzed and found that the expression of CCR6 was related to age and gender.There was no correlation between pathological type,but it was associated with neurological invasion of adenoid cystic carcinoma,suggesting that CCR6 may be one of the biological markers of nerve invasion in salivary adenoid cystic carcinoma.CCR6 and its ligand CCL20 may be related to salivary glands.Peripheral nerve infiltration and growth of adenoid cystic carcinoma are closely related.2.In vitro experiments,interfere with the expression of CCR6,have a significant inhibitory effect on human salivary gland adenoid cystic carcinoma cell proliferation,invasion and exercise capacity,CCL20 can significantly promote the movement of human salivary adenoid cystic carcinoma cells,CCR6 and its ligand CCL20 is closely related to the characteristics of peripheral nerve infiltration and growth of salivary adenoid cystic carcinoma. |
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