摘要
目的:探讨胶质瘤相关癌基因1(Gli1)在胃癌组织中的表达,阐明Gli1基因对胃癌细胞增殖和迁移能力的影响。方法:收集95例胃癌患者胃癌组织和癌旁组织,RT-qPCR法检测胃癌组织和癌旁组织中Gli1mRNA表达水平,免疫组织化学法检测胃癌组织和癌旁组织中Gli1蛋白表达水平。以人胃癌细胞株MKN28、BGC823和SGC7901为研究对象,胃永生化上皮细胞株GES-1作为对照,RT-qPCR法检测各细胞株中Gli1mRNA表达水平。将Gli1-siRNA转染BGC823细胞后,实验分为对照组、con-siRNA组和Gli1-siRNA组;RT-qPCR法检测各组细胞中Gli1mRNA表达水平,CCK-8检测细胞增殖情况,Transwell小室检测细胞迁移能力,Western blotting法检测各组细胞P27、基质金属蛋白酶2(MMP-2)和基质金属蛋白酶9(MMP-9)蛋白的表达水平。结果:胃癌组织中Gli1mRNA表达水平和蛋白阳性表达率明显高于癌旁组织(t=27.606,P<0.01;χ2=54.782,P<0.01)。在GES-1、MKN28、SGC7901和BGC823细胞中Gli1mRNA表达水平比较差异有统计学意义(F=86.341,P<0.01)。Gli1-siRNA组Gli1mRNA表达水平明显低于对照组和con-siRNA组(F=48.322,P<0.01)。Gli1-siRNA组胃癌细胞增殖率明显低于对照组和con-siRNA组(F=54.428,P<0.01)。Gli1-siRNA组胃癌细胞的迁移数低于对照组和con-siRNA组(F=257.788,P<0.01)。与对照组比较,con-siRNA组细胞中P27、MMP-2和MMP-9蛋白表达水平差异均无统计学意义(P>0.05);与con-siRNA组比较,Gli1-siRNA组细胞中P27蛋白表达水平明显升高(t=-3.776,P=0.020),MMP-2和MMP-9蛋白表达水平明显降低(t=3.479,P=0.025;t=5.487,P=0.005)。结论:Gli1在胃癌组织表达水平高于癌旁组织,抑制Gli1基因的表达能抑制胃癌细胞增殖和迁移能力。
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 and immortalized gastric epithelial cells GES-1 were cultured,GES-1 as a reference,and the expression levels of Gli1 mRNA 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;χ2=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.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2018年第1期63-67,共5页
Journal of Jilin University：Medicine Edition
基金
河北省科技厅科技支撑计划项目资助课题(152777130)
关键词
胶质瘤相关癌基因1
胃肿瘤
细胞增殖
细胞迁移
glioma-associated oncogene 1
stomach neoplasms
cell proliferation
cell migration
