| Objective: Gastric cancer is one of the most common malignant tumor in China,ranking third in the cancer cause of death.In China,most patients have been in advanced stage when they see a doctor,so a lot of treatments fail to obtain satisfactory curative effect,and until now,the mechanism of gastric cancer development is still unclear.CDK5RAP3 is the binding protein of the Cdk5 activator protein p35 and P39.Although CDK5RAP3 has been implicated in cancer progression,its expression and function have not been investigated in gastric cancer.At first this study investigated the correlation between CDK5RAP3 expression and development of gastric cancer,and analysised the relationship between CDK5RAP3 expression and clinicopathological parameters or prognosis.Secondly,through molecular biological experiments in vivo and in vitro,we investigated the relationship between CDK5RAP3 expression and gastric cancer cell proliferation,invasion and migration.Finally,this paper discussed the downstream gene of CDK5RAP3 gene mediating the development of gastric cancer and the potential molecular mechanisms of which.This study aims to investigate the role of CDK5RAP3 and the underlying molecular mechanisms in gastric cancer.Methods: 1.Using the immunohistochemical?qPCR and Western blotting method to detect CDK5RAP3 gene expression level in human gastric cancer samples,and to explore the correlation between CDK5RAP3 gene and clinical pathological characteristics or prognosis in gastric cancer patients.2.Using the packaged lentivirus to build the gastric cancer cell lines HGC-27,NUGC-3 and MGC-803 with stable overexpressed CDK5RAP3 and stable silenced CDK5RAP3.3.Through CCK8,cell counting method and plate clone formation experiment,discussing the relationship between CDK5RAP3 expression and gastric cancer cell proliferation.4.Through Transwell little room and scratch test,investigating the correlation between CDK5RAP3 gene expression and the invasion and migration of gastric cancer cells.5.By building the nude mice subcutaneously into tumor model,validating the influence of CDK5RAP3 in HGC turn-27 stabilizing gastric cancer cell lines forming subcutaneous tumor in nude mice;6.Using nuclear mass separation,confocal,immunofluorescence techniques to discuss the potential downstream target genes of CDK5RAP3 gene as well as the possible molecular mechanisms.Results: 1.CDK5RAP3 protein and mRNA expression levels in gastric cancer tissues were significantly lower than that in normal tissues.2.CDK5RAP3 genes were independent factors affecting the prognosis of patients with gastric cancer,and the 5-year survival rate of patients with low expression of CDK5RAP3(42.1%)was significantly lower than that of patients with high expression of CDK5RAP3(63.0%)(P<0.05).3.CDK5RAP3 gene expression mediated the gastric cancer cell proliferation,invasion and migration.The proliferation,invasion and migration ability of gastric cancer cell line with stable overexpression of CDK5RAP3 were decreased,while the gastric cancer cell line after stablely silencing CDK5RAP3 had the opposite result.4.Overexpression of CDK5RAP3 gene could reduce beta catenin protein expression,and CDK5RAP3 genes affected the change in function of gastric cancer cells depended on the ?-catenin protein expression.5.CDK5RAP3 influenced the expression of ?-catenin protein through inhibiting phosphorylation of GSK-3? Ser9 site level.6.The prognostic impact of CDK5RAP3 on the survival of patients with gastric cancer was dependent on the ?-catenin protein expression positioning.Conclusion: 1.CDK5RAP3 closely associated with gastric cancer development,which is a promising indicator to predict the prognosis of patients with gastric cancer;2.CDK5RAP3 can not only affect the gastric cancer cell proliferation,but also mediate the change of the gastric cancer cell invasion and migration ability.3.CDK5RAP3 can inhibit GSK-3? phosphorylation level while ?-catenin protein expression,and may be served as a potential target for the treatment of gastric cancer clinically. |
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