Role Of C1GALT1 In Gastric Carcinogenesis And Its Molecular Mechanism

Objective:To detect the expression of C1GALT1 in gastric cancer tissue and to observe its effect on malignant biological function by silencing C1GALT1 in MGC-803 gastric cancer cells.This study aimed to determine the role of C1GALT1 expression in the evolution of gastric cancer.Methods:1.Confirmation of the specificity of C1GALT1 reaction by fluorescence quantitative PCR.2.The expression of C1GALT1 mRNA and C1GALT1 protein in 46 cases of gastric cancer tissues and corresponding paracancerous tissues was detected by RT-PCR and Western blot respectively.3.The expression of C1GALT1 mRNA and C1GALT1 protein in gastric cancer cell lines SGC-7901,MGC-803,BGC-823 and normal gastric mucosal cell line GES-1 were detected by RT-PCR and Western blot.4.C1GALT1-siRNA sequence was transfected into MGC803 cells,C1GALT1-siRNA transfection group,empty plasmid vector group and blank control group were established.Western blot was used to detect the expression of C1GALT1 protein in each group.MTT assay,in vitro cell migration assay,In vitro cell invasion assay clear MGC803 cells proliferation,migration and invasion ability changes.Results: 1.Fluorescent Quantitative PCR The C1GALT1 dissolution profile produced after the reaction showed a single peak near 90 degrees indicating a single,non-specific amplification product of the PCR product as well as primer-dimers.2.The results of RT-PCR showed that:(1)36 cases of C1GALT1 mRNA expressed more in gastric cancer tissues than in matched normal tissues(foldchange = 3.91 ± 3.61);10 cases had no difference(foldchange = 0.95 ± 0.34).Chi(2)-square analysis showed χ2 = 2.504,P = 0.114,the results showed that C1GALT1 mRNA expression in metastatic and non-metastatic gastric cancer was no significant difference.(3)Spearman bivariate correlation analysis of C1GALT1 mRNA expression and clinical stage of gastric cancer,the results showed that r = 0.761,P<0.001.The results showed that the expression of C1GALT1 mRNA positively correlated with the development of gastric cancer.(4)The expression of C1GALT1 mRNA in SGC-7901,BGC-823 and MGC-803 gastric cancer cell lines was significantly higher than that in normal gastric mucosa cell line GES-1(foldchange> 3.2,P<0.001).3.The results of Western blot showed that:(1)The expression of C1GALT1 protein in 37 gastric cancer tissues was higher than that in the adjacent tissues(foldchange = 3.78 ± 1.28,P<0.001),and no change in 9 cases(foldchange = 0.85 ± 0.15).(2)The use of chi-square analysis to compare the expression of C1GALT1 protein in metastatic and non-metastatic gastric cancer tissues,the results showed χ2 = 6.08,P = 0.014,the results have significant differences.(3)Spearman bivariate correlation analysis of C1GALT1 protein expression and clinical staging of gastric cancer,the results showed that,r = 0.329,P<0.026.The results showed that the expression of C1GALT1 protein positively correlated with the development of gastric cancer.(4)C1GALT1 protein expression in SGC-7901,BGC-823,MGC-803 gastric cancer cell lines than the normal gastric mucosal cell line GES-1 specific increase(foldchange> 1.73,P<0.05).4.MGC803 cells transfected with specific C1GALT1-siRNA sequence,FAM fluorescence prompted the transfection rate of more than 90%.5.C1GALT1-siRNA transfection group,plasmid empty vector group and blank control group,C1GALT1 protein expression in the transfection group by Western blot showed that the grayscale was 0.432 +0.0413,significantly lower than Ctrl-siRNA group(0.812 +0.0221)And untransfected group(0.853 + 0.0132)(P <0.05).6.MTT experiment,in vitro cell migration experiment and in vitro experiment invasion assay: Compared with the control group and empty vector group,MGC803 cells transfected with C1GALT1-siRNA significantly reduced the ability of proliferation,migration and invasion(P <0.05).Conclusion:1.The expression of C1GALT1 mRNA in gastric cancer tissues is significantly higher than that in adjacent tissues,and may be positively correlate with the clinical stage of gastric cancer.2.The expression of C1GALT1 protein in gastric cancer tissues is significantly higher than that in adjacent tissues,and is positively correlate with the clinical stage of gastric cancer.3.C1GALT1 may promote the biological behaviors such as proliferation,migration and invasion of gastric cancer cells.