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.
ABSTRACT
Objective To explore the relationship and clinical significance of Gli1 expression in peripheral blood and cancer tissues of gastric.Method 95 gastric cancer patients were recruited as surgery group,and 40 recurrent gastric cancer patients were recruited as recurrent group.50 normal subjects were recruited as control group.Paraffin specimens of gastric cancer tissues and gastric mucosa adjacent to cancer were collected to test expression of Gli1 protein.ELISA was applied to test Gli1 in peripheral blood.Results Levels of Gli1 in peripheral blood of preoperative surgery group were higher than in control group [(9.21 ± 1.37) ng/ml vs.(0.79 ±0.18) ng/ml,t =43.193,P ＜0.001].Post-up Gli1 decreased significantly to the level of control group (t =0.722,P =0.472).Levels of Gli1 in peripheral blood of recurrent group were higher than that in preoperativesurgery group (t =14.575,P ＜ 0.001).Gli1 protein in cancer tissues was higher than that in adjacent gastric mucosa (t =33.471,P ＜0.001),and Gli1 protein in cancer tissues of surgery group was higher than that in cancer tissues of recurrent group (t =8.808,P ＜0.001).Conclusion In postoperative gastric cancer patients increased Gli1 level in peripheral blood and cancer tissues predicts recurrence.
ABSTRACT
Background and purpose:miRNA plays important roles in tumorigenesis. It has been reported that many kinds of serum miRNA serve as markers for tumor diagnosis and screening. This study aimed to detect the expression of serum miRNA-31 (miR-31) in colorectal cancer patients and to explore the effect of miR-31 on cell proliferation, apoptosis and cell cycle distribution. Methods: The expressions of miR-31 in 40 cases of colorectal cancer serum and 35 cases of the healthy control were examined by real-time lfuorescent quantitative polymerase chain reaction (RTFQ-PCR). The correlation between miR-31 expression and clinicopathological features of colorectal cancer (including age, gender, depth of inifltration, lymph node metastasis, clinical stage) were further analyzed. The miR-31 mimics, inhibitor and miR-control (negative control) were transfected into HCT116 cells. The effect of miR-31 on cell proliferation was evaluated by CCK-8 method. Flow cytometry was used to examine the change of cell apoptosis and cell cycle. Results:Relative expression of serum miR-31 was signiifcantly increased in cancer patients compared with healthy controls (P<0.01). Expression of serum miR-31 was higher in poorly differentiated carcinoma than that in well or moderately differentiated carcinoma (P<0.05). No correlation was found between serum miR-31 expression and other clinicopathological variables. CCK-8 assay showed that after transfection with miR-31 mimics, the cell proliferation was increased, compared with miR-31 inhibitor and negative control group. Meantime, the apoptotic cell number was signiifcantly decreased, particularly in late apoptosis. The cell number of G1 stage was remarkably increased in miR-31 inhibitor group, compared with miR-31mimics and negative control group. Conclusion:The expression of serum miR-31 is higher in colorectal cancer. miR-31 can promote cell proliferation and inhibit the apoptosis of HCT116 cells. It might be a potential biomarker for colorectal cancer.