Expression of Gli1 in gastric carcinoma tissue and its effect on biological characteristics of gastric carcinoma cells / 吉林大学学报(医学版)

ABSTRACT

Objective:To study the expression of glioma-associated oncogene 1 (Gli1 in gastric carcinoma tissue,and to explore the effects of Gli1 on the proliferation and migration abilities of gastric carcinoma cells.Methods:A total of 95 cases of human gastric carcinoma tissue and paracancerous tissue were collected.Immunohistochemistry was used to detect the expression levels of Gli1 protein in gastric carcinoma tissue and paracancerous tissue;Real-time PCR method was used to detect the expression levels of Gli1 mRNA in gastric carcinoma tissue and paracancerous tissue.The human gastric cancer cell lines MKN28,BGC823,SGC7901 andimmortalized gastric epithelial cells GES-1 were cultured,GES-1 as a reference,and the expression levels of Gli1mRNA in cell lines were detected by RT-qPCR.The Gli1-siRNA and con-siRNA were transfected into the gastric carcinoma cell line BGC823,and control group,con-siRNA and Gil1-siRNA group were set up.The expression levels of Gli1 mRNA in the cells in various groups were detected by RT-qPCR;CCK8 was used to detect the proliferation of cells;Transwell migration assay was used to detect the cell migration ability.Western blotting was used to detect the expression levels of the P27,matrix metalloproteinase-2 (MMP-2),and MMP-9 proteins in the cells in various groups.Results:The positive expression rates of Gli1 mRNA and protein in gastric carcinoma tissue were significantly higher than those in paracancerous tissue (t=27.606,P＜0.01;x2=54.782,P＜0.01).There were significantly differences in the expression levels of Gli1 mRNA between GES-1,MKN28,SGC7901,and BGC823 cell lines (F =86.341,P ＜0.01).The expression level of Gli1 mRNA in Gli1-siRNA group was significantly lower than those in control and con-siRNA groups (F=48.322,P＜0.01).The proliferation rate of gastric carcinoma cells in Gli1-siRNA group was significantly lower than those in control and con-siRNA groups (F=54.428,P＜0.01).The results of Transwell migration assay showed that the number of migration cells in Gli1-siRNA group was significantly lower than those in control and con-siRNA groups (F=257.788,P＜0.01).The Western blotting results showed that the expression levels of P27,MMP-2,and MMP-9 proteins in Gli1-siRNA group had no significant differences compared with control group (P＞0.05).Compared with con-siRNA group,the expression level of P27 in the cells in Gli1-siRNA group was significantly increased (t=-3.776,P=0.020),and the expression levels of MMP-2 and MMP-9 proteins were significantly dereased (P =3.497,P =0.025;t=5.487,P=0.005).Conclusion:The expression level of Gli1 in gastric carcinoma tissue is higher than that in paracancerous tissue,and the inhibition of the expression of Gli1 gene could inhibit the cell proliferation and migration.