Study On The Effect Of SiRNA Silencing FGFR3 Gene Expression On Bone Metastasis Of Nude Mice Induced By Lung Cancer SPC-A-1BM Cells

Objective:To investigate the effect of the silencing of Fibroblast Growth Factor Receptor-3（FGFR3）gene on the SPC-A-1BM cells of human lung adenocarcinoma with high bone metastasis and its effect on bone metastasis in nude mice.Methods:The experiment was divided into three groups:1 groups were experimental group（FGFR3 specific siRNA interference group）,the 2 group was negative control group（FGFR3 non-specific negative control siRNA interference group）,and the 3 group was blank control group（no siRNA interference group）.The transfection of high bone metastatic human lung adenocarcinoma cell line（SPC-A-1BM cell）was mediated by Lipofectamine^TM2000（Lipo2000）mediated by nucleic acid transfection reagent,liposome Lipofectamine^TM2000（Lipo2000）;Real time Real-time quantitative polymerase chain reaction（Real-time PCR）was used to detect mRNA expression level of FGFR3gene after transfection;Protein immunoblotting（Western-Blot）was used to detect the expression level of FGFR3 gene before and after transfection;The model was established by tail vein injection in nude mice,and the lung cancer SPC-A-1BM cells were transfected into nude mice respectively;The hematoxylin eosin staining（HE staining）was used to make some tissue pathological sections of nude mice and to make clear the pathological diagnosis of bone metastasis of lung cancer SPC-A-1BM cells in nude mice,and to differentiate and compare the cases of bone metastases in each group.Results:The transfection efficiency of siRNA using Lipo2000 as carrier was more than70%for lung adenocarcinoma SPC-A-1BM cells;The Real-time PCR experiment showed that the expression of FGFR3 gene in the 1 groups of the experimental group was obviously inhibited,the inhibition rate was about 71%,and the 2 groups of the negative control group were compared with the FGFR3 gene expression in the blank control group3 groups,and the obvious downregulation was seen,and the single factor variance analysis was used（P<0.01）;Western-Blot experiments showed that for FGFR3 protein,the expression of protein in the 1 groups of the experimental group was significantly lower than that of the negative control group 2 and the blank control group 3,and the difference between the band gray values was statistically significant（P<0.01）;After modeling nude mice in each group through tail vein injection,2 groups of negative control groups and 3 groups of blank control group could be seen that about 50%-60%of nude mice could see different degrees of bone metastases;Histopathological pathology,wax block making,slice and HE staining experiment showed that the 1 groups of the experimental group compared the negative control group 2 groups and the blank control group 3 groups of nude mice,the number of nude mice with lung cancer bone metastasis and the bone metastases in nude mice decreased significantly,the difference was statistically significant（P<0.01）.Connclusion:The transfection of FGFR3 gene transfected with SPC-A-1BM cells by Lipo2000 as a carrier mediated FGFR3 gene can obviously inhibit the invasion ability of lung cancer cells and reduce the occurrence and development of bone metastasis;This experiment provides a theoretical basis for the treatment of FGFR3 gene silencing of bone metastases,and provides new therapeutic targets for the treatment of bone metastasis of lung cancer.