The Mechanism Research Of CDK5RAP3 In The Carcinogenesis And Metastasis Of Gastric Cancer

Object:Gastric cancer is the second cause of death in the disease,in china,it is also one of the major malignant tumor,and in china gastric cancer is often with a high incidence,high transfer rate and so on.As a kind of serious damage to human health disease,invasion and metastasis is main lethal factor of gastric cancer.But so far,the transfer mechanism of gastric cancer has not been fully elucidated,the mechanism research in the metastasis of gastric cancer still required deep-going research,and it is particularly important.Our previous study found CDK5RAP3 plays the role of tumor suppressor in gastric cancer,and CDK5RAP3 expression is closely related to the prognosis of patients with gastric cancer.In this study we want to investigate the relationship between the CDK5RAP3 and the metastasis of gastric cancer,also to reveal the underlying mechanisms of invasion and metastasis in gastric cancer and provide new targets for gastric cancer treatment and intervention methods.Methods:1.Using the packaged lentivirus to build the gastric cancer cell lines HGC-27 and NUGC-3 with stable overexpressed CDK5RAP3 and stable silenced CDK5RAP3;2.Through Transwell little room and scratch test,investigating the correlation between CDK5RAP3 gene expression and the migration of gastric cancer cells;3.We investigated the effects of CDK5RAP3 on metastasis in the stable HGC-27 cells through inoculating HGC-27 and its control cells into the caudal vein of nude mice to build the gastric cancer models;4.Using CDK5RAP3 plasmid intervention NUGC-3 cells,then using Western blot experiments to discuss the influence of the downstream mechanism due to CDK5RAP3 expression;5.In the stable overexpression and stable knockout CDK5RAP3 gastric cancer cell lines HGC-27 and NUGC-3,using q PCR and Western bloting technology to observe the changes of EMT Signaling pathway due to the expression changes of CDK5RAP3.Results:1 Using the packaged lentivirus has been successfully constructed the gastric cancer cell lines HGC-27 and NUGC-3 with stable overexpressed CDK5RAP3 and stable silenced CDK5RAP3,and observed CDK5RAP3 protein expression will affect the morphology of the gastric cancer cell lines;2.Using transwell chambers experiment observed CDK5RAP3 protein overexpression inhibited gastric cancer cell migration ability,and decreased CDK5RAP3 protein expression will promote the gastric cancer cell migration ability;3.In vivo experiments,compared with negative CDK5RAP3 control group,downregulation of CDK5RAP3 dramatically increased the invasive capability in HGC-27 cells(p <0.01);4.CDK5RAP3 protein expression changes will affect the EMT signaling pathway mainly protein E cadherin,N-cadherin,ZO-1 and Vimentin transcription level.5.CDK5RAP3 protein expression changes will affect the EMT signaling pathway mainly protein E cadherin,N-cadherin,ZO-1 and Vimentin protein expression.Conclusions:1.CDK5RAP3 gene can change the gastric cancer cell morphology;2.CDK5RAP3 gene inhibits gastric cancer cell migration ability;3.CDK5RAP3 genes can influence the EMT signaling pathway main protein E cadherin,N-cadherin,ZO-1 and Vimentin transcription level and protein expression level.