The Effect Of Si Nomenine On The Downregulation Of ? 7nAChR And Anti-Lung Tumor

Objective:In this study,model of mice bearing Lewis lung cancer cell and human non small cell lung cancer cell line A549,as the object were study to observe the effect of sinomenine on lung cancer proliferation and the expression of a 7nAChR.So as The effect of sinomenine on the lung cancer cells proliferation and the relationship between nAChR.How sinomeninethe activate the MAPK pathway in vitro,further,the regulatory mechanism of sinomenine on the expression of a 7nAChR in lung cancer cells was explore..Method:1.Effects of sinomenine on model of mice bearing Lewis lung cancer cell:mouse lung cancer cell line Lewis was cultured in vitro.the effect of sinomenine on the proliferation of Lewis cell was detected.Model of Lewis cell in C57BL/6 mice were established with subcutaneous injection of lewis for 7 days:sinomenine high-dose group(160mg/kg),sinomenine low-dose group(80mg/kg),DDP group(lmg/kg),NNK group(1.5mg/kg).The weight of transplanted tumor were detected after administration for 14d,TGF-? and VEGF in serum were detected by ElISA;The expression of a 7nAChR in transplanted tumor tissues were deteced by Western blot.2.The mechanism of sinomenine on the proliferation of A549 cell lines in vitro:MTT was used to detected the effect of sinomenine in different concentrations on the proliferation of A549,NNK,Ber as control observation.Then to observe the effect of nAChR antagonist mecamylamylamine and mAChR antagonist atropine on sinomenine against A549 cell proliferation.Effect of sinomenine on the expression of expression of ERK1/2 and P-ERK1/2 were detected by Western blot.3.Effects of sinomenine,NNK and Ber on the expression of a 7nAChR?Egr-1,SP-1 and TTF-1 mRNA and protern in human lung cancer cell line A549 were detected by RT-PCR and Western blot.Results:1.Sinomenine can inhibit the proliferation of Lewis cells in vivo,and in a dose-dependent manner.Sinomenine can inhibit model of mice bearing Lewis lung cancer cell,reduce TGF-p and VEGF in serum;Cisplatin can inhibit model of mice bearing Lewis lung cancer cell,reduce TGF-? and VEGF in serum;NNK can promote model of mice bearing Lewis lung cancer cell,increase the content of VEGF,TGF-? in serum.Sinomenine can reduced levels of a 7nAChR protein in model of mice bearing Lewis lung cancer cell.Cisplatin had no obvious effect on the level of a 7nAChR protein in model of mice bearing Lewis lung cancer cell.NNK can increase the level of a 7nAChR protein in model of mice bearing Lewis lung cancer cell.2.Sinomenine can inhibit the proliferation of A549 cells in vivo.the effect Sinomenine can be antagonized by mecamylamine;Ber can inhibit the proliferation of A549,the effect cannot be antagonized by mecamylamine or atropine;NNK can promote the proliferation of A549,the effect Sinomenine can be antagonized by mecamylamine;ACh can promote the proliferation of A549,the effects can be antagonized by atropine.Sinomenine can reduced the ratio of P-ERK1/2 in A549.3.Sinomenine can reduce the level of a 7nAChR mRNA and protein in A549.Ber had no significant effect on the level of a 7nAChR mRNA and protein in A549.NNK can increase the level of a 7nAChR mRNA and protein.Sinomenine can increase Egr-1 mRNA and protein levels in human lung cancer cells A549,decrease SP-1,TTF-1 mRNA and protein,ber had no obvious effect on Egr-1,SP-1,TTF-1mRNA and protein levels,NNK can reduce Egr-1 mRNA and protein,increase SP-1,TTF-1 mRNA and protein.Conclusion:Sinomenine can inhibit lung cancer and inhibit the expression of a 7nAChR in lung cancer.Sinomenine can inhibit the lung cancer cell proliferation by nAChR pathway ang downstream pathway.Sinomenine can down regulate the expression of a 7nAChR in lung cancer cells,which may be associated with up regulation of Egr-1 and down-regulation of SP-1 and TTF-1.