| Objective:To further study the mechanism of mi R-711 inhibiting the occurrence and development of gastric cancer,we will observe the effect of mi R-711 on Nude mice and the expression of Sp1,E-cadherin and Vimentin in SGC-7901 Human gastric cancer cell based on previous studies.Methods :1)Conclusion lentiviral vector carrying miR-711(experimental group)and non-transfection group(negative control group),infecting SGC-7901 cells,Fluorescence microscopy was used to observe the transfection efficiency.2)SGC-7901 cells transfected with mi R-711 or control vector were injected subcutaneously into nude mice.Tumor growth was montored by calipers.The volume of the tumor was calculated by the formula:tumor volume =(width)2 × length/2.Portraying the transplantation tumor' growth chart diagram.The mice were sacrificed after 4-6weeks,tumors were stripped and weighed up:(1)The organization form of tumor was observed after HE staining in each group.(2)Real-time PCR was used to detect the expression of miR-711 in tumor tissues.(3)The expression of EMT related proteins: E-cadherin ?Vimentin which come from xenograft tumor proteins were assayed by immunohistochemical staining.(4)The expression of Sp1,Vimentin and E-canherin protein in nude mice was detected by Western blotting.Results:1)The transfection efficiency of SGC-7901 cells observed under fluorescence microscope was more than 70%.2)We successfully established the nude mice transplanted tumor model with overexpression of miR-711 gene and control vector in human gastric cancer SGC-7901 cells.The results showed that Compared with the negative control group,the expression of tumor volume and tumor weight in the overexpression group were significantly lower than those in the negative control group.microscope lens showed : nude mice xenograft tumor of the experimental group had less atypia and inflammatory than the negative control group.(P <0.05).3)Real-time PCR results showed that the expression level of miR-711 in overexpression group was significantly higher than that in negative control group.4)Immunohistochemical analysis showed that the epithelial marker E – cadherin had high expression in experimental group compared with the control group,while the expression of Vimentin was less than control group.the difference was significant(P < 0.05).5)Compared with the negative control group,the expression of E-cadherin protein in the miR-711 overexpression group was increased and the expression of Vimentin and Sp1 protein was decreased.There was significant difference(P <0.05).Conclusion:1?MiR-711 could inhibit the tumorigenic ability of SGC-7901 human gastric cancer cells in nude mice and the expression of related proteins of EMT. |
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