Study On Effect Of M3 In Regulating Nerve Invasion Of Cholangiocarcinoma

Objective:To investigate the role of M receptor agonists and antagonists on regulating the perineural invasion of cholangiocarcinoma and to explore the possible mechanism of this action,in order to provide new approaches to the drug treatment of cholangiocarcinomaMethods:1.FRH0201 and RBE cell culture2.To detect the expression of M3 receptor gene in the cholangiocarcinoma cells By RT-PCR and WESTERN BLOT.3.Adenovirus m3-siRNA transfected cholangiocarcinoma cells4.To simulate the conditiong of the cholangiocarcinoma's neural invasion by means of the co-colture model of DRG/nerve cells and carcinoma cells.5.the invasion process of the human bile duct carcinoma cell line FRH0201 and RBE were observed.After 72 hours,find out the carcinoma cells which were sticked to the neurite.The data were analyzed with SPSS 16.0 statistical software.Results:1.By RT-PCR,and WESTERN BLOT,we realise that M3 receptor gene in RBE cells is relatively highly expressed,while the M3 receptor gene of FRH0201 cells is relatively lowly expression;2.M3 receptor gene was successfully transfected these two cholangiocarcinoma cells and stable expression.3.In the co-colture model,after 72 hours,find out the carcinoma cells which were sticked to the neurite.In cevimeline group the number of carcinoma cells which invaded the neurite had a significant difference compared with the control group and the cevimeline+4-DAMP group,P<0.05;The number of carcinoma in 4-DAMP group had no difference compared with the control guoup,P>0.05.Conclusion:Cholangiocarcinoma cells may express M3 muscarinic acetylcholine receptors,receptor agonists can promote nerve invasion of cholangiocarcinoma.It provides experimental basis for the incidence of perineural invasion and future development.