In Vivo Experimental Study On ShRNA-mediated Silencing Of FAK Gene Inhibits Growth And Metastasis Of Gastric Cancer

Object:To investigate the tumor growth and metastasis in stomach cancer cells transfectedwith silence FAK genes in vivo of nude mouse.Methods:Cultured the gastric cancer cell line SGC-7901,the cells were classified as threegroups: transfected group was handled with FAK-shRNA which recombinated intoLentivirus vector; negative control group was handled with blank shRNA,recombinated into Lentivirus vector; the blank control group was kept originalstate.screening for stability transfected cell line under the condition of G418concentration in600ug/ml; The amount of FAK protein which had been expressedfrom three different groups of cells was detected by means of Western blot. The nudemice were divided into six groups randomly, and three groups of cells were passagedfor a long time can obtain a great amount of each type. Transplanted those threegroups cells to nude mice in order to creating the model of human gastric cancer innude mouse stomach and subcutaneous cancer. Then their primary tumor growth andmetastasis were observed. Using RT-qPCR and immunohistochemical stainingmethods detected the expression of mRNA and protein from gene of FAK.Results:1. Comparing with negative control group and blank control group, focal adhesionkinase detected by the Western blot test in transfected group was obviously reduced.2. The weight of transplant subcutaneous tumor in nude mouse: there weresignificant differences between transfected group (1.474±0.9840g) and negativecontrol group(3.134±0.3299g),P <0.05; the same conclusion can be obtained from the contrast between transfected group and blank control group(2.687±0.3827g),P<0.05.Negative control group and blank group compared to eachother had not found statistics meaning. Metastasis in the original tumor model of nudemouse: cases of gastric carcinoma with hepatic metastasis were found1in transfectedgroup，5in negative control group and6in blank control group; there were4nudemice with abdominal cavity metastasis in negative control group and blank controlgroup respectively while no metastasis was found in transfected group. According todata statistics，significant differences could be found among three groups.3. Using RT-qPCR detection and immunohistochemical staining methodswith thesubcutaneous tumor, the mRNA expressed from gene of FAK and FAK protein in thetransfected group is obviously lower than the negative control group and the blankcontrol group..Conclusions:1. FAK-shRNA can significantly suppress FAK expression in gastric cancer SGC-7901cell line.2. FAK-shRNA can reduce the transcription of mRNA from FAK gene and the contentof FAK protein in nude mice carcinoma obviously.3. The silencing of FAK resulted in the inhibition of gastric tumor metastasis in nudemice.