Effects And Mechanism Of Dio On Invasion And Metastasis Of Gastric Cancer

Part Ⅰ The effect of dio on the proliferation and migration of gastric cancer cellsObjective:To investigate the effect of Dio on proliferation and migration of gastric cancer cells.Methods:SGC-7901 and AGS gastric cancer cell lines were selected,(1)it was divided into Ctrl group and Dio group(0.5,1,2,4,8 μM),and the effect of Dio on the proliferation of gastric cancer cells was determined by MTT.(2)It was divided into Ctrl group and Dio group(0.5,1,2μM),scratch test and Transwell were used to detect the influence of Dio on the gastric cancer cells migration.(3)It was divided into Ctrl group and Dio group(0.5,1,2 μM),Western-Blot was used to detect the expression of metastasis-related proteins such as MMP-2 and MMP-9 to explore the effect of Dio on the migration of gastric cancer cells.Results:(1)The result of MTT showed that Dio could inhibit the proliferation of gastric cancer cells in a time-and concentration-dependent manner(P<0.05);(2)The result of scratch test showed that the higher the concentration of Dio,the lower the scratch healing rate(P<0.05);(3)The results of Transwell showed that the higher the concentration of Dio,the less the migration of the cells.(3)The result of Western-Blot showed that Dio could inhibit the expression of metastasis-related proteins such as MMP-2 and MMP-9.Conclusion:Dio can inhibit the proliferation and migration of gastric cancer cells.Part Ⅱ.Investigate the effect of Dio on epithelial-mesenchymal transition of gastric cancer cellsObjective:To investigate the effect of Dio on epithelial-mesenchymal transition of gastric cancer cells.Methods:SGC-7901 and AGS gastric cancer cell lines were selected and it was divided into Ctrl group and Dio group(0.5,1,2 μM).The expression of epithelial-mesenchymal transition-related proteins such as E-cadherin,N-cadherin,Vimentin and Snail-1 were detected by Western-Blot to explore the effect of Dio on epithelial-mesenchymal transformation of gastric cancer cells.Results:The result of Western-Blot showed that Dio could up-regulate the expression of E-cadherin,while down-regulate the expression of N-cadherin and Vimentin and Snail-1(all P<0.05).Conclusion:Dio can inhibit the epithelial-mesenchymal transition of gastric cancer cells.Part Ⅲ.Investigate the expression of Cx43 in gastric cancer and its correlation with clinicopathological features and prognosisObjective:To analyze the correlation between the expression of connexin 43 and clinicopathological features and prognosis in gastric cancer..Methods:From March 2015 to February 2017,a total of 68 gastric cancer patients who have underwent radical gastrectomy in Luoyang Central Hospital were collected.The clinicopathological data were prospectively recorded for retrospective analysis and followed up to August 2020.The expression of Cx43 in tissues was detected by immunohistochemistry,and the correlation between the expression of Cx43 and clinicopathological features was analyzed.Kaplan-Meier was used to construct the total survival curve and disease-free survival curve of patients with gastric cancer.Besides,the independent influencing factors of prognosis of gastric cancer patients were evaluated by Cox regression analysis.Results:The expression of Cx43 in gastric cancer(51.47%)was lower than that in paracancerous normal tissues(95.58%P<0.001).The expression of Cx43 was significantly correlated with the maximum diameter of tumor(P=0.004)and the depth of tumor invasion(P=0.012).In addition,Cox regression analysis showed that the expression of Cx43,the maximum diameter of tumor,the degree of tumor differentiation and the depth of invasion were independent factors affecting the prognosis of patients with gastric cancer.Conclusion:The expression of Cx43 is low in gastric cancer,and the low expression of Cx4 is negatively correlated with the prognosis of gastric cancer patients.The expression of Cx43 can be used as a potential prognostic index for patients with gastric cancer.Part Ⅳ Effect of Dio on Cx43 expression and possible mechanism of inhibiting gastric cancer invasion and metastasisObjective:To explore the effect of Dio on Cx43 and the possible mechanism of Dio up-regulating Cx43 and inhibiting epithelial-mesenchymal transformation of gastric cancer cells.Methods:AGS gastric cancer cell lines was selected.(1)It was divided into Ctrl group,Dio group(0.5,1,2 μM)and positive control group.The expression of Cx43 was detected by Western-Blot to explore the effect of Dio on the expression of Cx43 in gastric cancer cells.(2)It was divided into Ctrl group,Dio group(0.5,1,2 μM)and positive control group.The expression of P-PI3K,P-Akt,P-mTOR and PI3K,Akt,mTOR was detected by Western-Blot to explore the effect of Dio on PI3K/Akt signalling pathway of gastric cancer cells.Results:(1)The higher the concentration of Dio,the higher the expression of Cx43 in gastric cancer cells.(2)The higher the concentration of Dio,the lower the expression of P-PI3K,P-AKT and P-mTor,and the higher the concentration of Dio,the lower the ratio of P-AKT/Akt.Conclusion:Dio may inhibit epithelial-mesenchymal transition and the invasion and metastasis of gastric cancer by up-regulating the expression of Cx43 and inhibiting the activation of PI3K/Akt signalling pathway.