Thermo-chemotherapy Changes Gene Expression In Gastric Cancer Cells Via Down-regulate EIF5A2 To Inhibit Invasion And Metastasis Of Gastric Cancer

Part I Correlation between EIF5A2 factor and proliferation,invasion and metastasis of gastric cancer cells Gastric cancer is one of the most common malignant tumors in our country.Most of the patients have undergone peritoneal metastasis when diagnosed,and the traditional treatment is very poor.Eukaryotic Initiation Factor 5A2,EIF5A2 is a highly conserved proteinin in eukaryotes,which is highly expressed in colorectal cancer,ovarian cancer and other malignant tumors.It could up-regulate expression of proliferation related genes Cyclin D1 and Cyclin D3 to promote tumor cell proliferation,and up-regulate invasion and metastasis genes MTA1 and c-myc,up-regulate epithelial mesenchymal transition genes Vimentin,down-regulate E-cadherin to induce epithelial mesenchymal transition and promote invasion and metastasis through of tumor cells.Objective:To explore the expression of EIF5A2 in human gastric cancer cells and its correlation with proliferation,invasion and metastasis of gastric cancer cells.Method:MKN28 cells with high expression of EIF5A2 factor was respectively transfected with EIF5A2 siRNA and control si RNA,and MKN45 cells with low expression of EIF5A2 factor was transfected with EIF5A2 high expression plasmid and blank plasmid.CCK-8 was applied to determinate the proliferation activity of gastric cancer cells;Transwell method,cell scratch test to test gastric cancer cells' invasion and migration ability;Real-time quantitative PCR(qRT-PCR)and Western Blot to detect the expression of EIF5A2,Cyclin D1,Cyclin D3,Vimentin,E-cadherin,MTA1 and c-myc.Result:Compared with MKN28-CTR group,the expression of EIF5A2 factor was down-regulated,as well as proliferation-related genes Cyclin D1 and Cyclin D3,and metastasis associated genes MTAl and c-myc,epithelial mesenchymal transition gene Vimentin,while the expression of E-cadherin was up-regulated in MKN28-siEIF5A2 group.And the proliferation,invasion and migration ability of MKN28 gastric cancer cells were restrained.However,compared with MKN45-CTR group,the expression of EIF5A2 factor was significantly up-regulated,as well as proliferation-related genes Cyclin D1 and Cyclin D3,and metastasis associated genes MTAl and c-myc,epithelial mesenchymal transition gene Vimentin,while the expression of E-cadherin was down-regulated in MKN45-EIF5A2 group.And the proliferation,invasion and migration ability of MKN45 gastric cancer cells were enhanced.Conclusion:The expression level of EIF5A2 factor in gastric cancer cells were positively correlated with the proliferation,invasion and metastasis ability of gastric cancer cells.EIF5A2 factor promote proliferation and invasion and metastasis of gastric cancer cells by up-regulating expression of proliferation,invasion and metastasis related genes.Part II The effect of thermo-chemotherapy on EIF5A2 factor and proliferation,invasion and metastasis gene expression and the correlation with invasion and metastasis of gastric cancerObjective: To research the effect of thermo-chemotherapy on the expression of EIF5A2 gene and invasion and metastasis related genes of gastric cancer cells,and the correlation with invasion and metastasis of gastric cancer.Methods: Cell experiment: MKN28 and MKN45 gastric cancer cell lines were respectively divided into three groups:(1)control group,Mock group(2)hyperthermia therapy group,HT group(43?),(3)chemotherapy group,CT group(L-OHP)(4)hyperthermia and chemotherapy group,HT+CT group(43?+L-OHP).CCK-8 was applied to detect the proliferation activity of gastric cancer cells;Transwell method,cell scratch test to detect the invasion and migration ability;q RT-PCR and western blot to detect expression of EIF5A2,Cyclin D1,Cyclin D3 and Vimentin,E-cadherin,MTA1,c-myc genes.Animal model experiment: MKN28,MKN45 gastric cancer cells were subcutaneous injected to the left groin to build xenograft tumor model.The mice were random Ly divided into 3 groups:(1)control group,Mock group(2)hyperthermia therapy group,HT group(43?),(3)chemotherapy group,CT group(L-OHP)(4)hyperthermia and chemotherapy group,HT+CT group(43?+L-OHP).Mice were treated with cervical dislocation after treatments.Tumor was resected completely,then observe necrosis of tumor tissues;q RT-PCR,western blot and immunohistochemical method to detect the expression of EIF5A2 factor,Cyclin D1,Cyclin D3 and Vimentin,E-cadherin,MTA1,c-myc genes.Result: Cell experiment: In MKN28 and MKN45 gastric cancer cells,compared with Mock group,the expression of EIF5A2 factor,proliferation relatied genes Cyclin D1 Cyclin D3 were down-regulated,and the proliferation ability of gastric cancer cells were significantly inhibited(p<0.05).The invasion and metastasis related genes MTA l,c-myc were significantly down-regulated,epithelial mesenchymal transition genes Vimentin down-regulated,E-cadherin up-regulated in MKN28 cells in HT,CT and HT+CT group,the difference were statistically significant(p<0.05).And the inhibite effect of proliferation,invasion and migration ability of gastric cancer cells were most significant in HT+CT group(p<0.05).Animal model experiment: The growth rate of tumor of mice in HT,CT and HT+CT group slowed down and showed different degree of necrosis,and the number of tumor necrosis in HT+CT group were more significant(P<0.05).The results of q RT-PCR,western blot and immunohistochemical method revealed that the expression of EIF5A2 factor,Cyclin D1,Cyclin D3 and invasion and metastasis related genes MTA l,c-myc were down-regulated,and the expression of Vimentin was down-regulated,E-cadherin up-regulated in HT,CT and HT+CT group,compared with Mock group,the difference were statistically significant(p<0.05),and the change of genes expression levels in HT+CT group were significantly higher than other groups(P<0.05).Conclusion: Thermo-chemotherapy can significantly inhibit the proliferation,invasion and migration ability of MKN28 and MKN45 gastric cancer cells.Thermo-chemotherapy inhibits the proliferation ability of gastric cancer cells by down-regulating the expression of EIF5A2 factor and Cyclin D1,Cyclin D3,and inhibits invasion and metastasis by down-regulating the expression of invasion and metastasis related genes MTA l,c-myc and Vimentin,up-regulating the expression of E-cadherin.