| Part Ⅰ:Analysis of the relationship between miR-532-3p expression and clinicopathological characteristics and β-catenin/Cyclin D1 in gastric cancerObjective:To investigate the expression and clinicopathological characteristics of miR-532-3p in gastric cancer β-catenin/Cyclin D1 relationship.Methods:All samples were collected from the pathological tissues and paracancerous tissues of 80 patients who underwent gastric cancer surgery in our hospital from July 2018 to July 2020.60 pathological tissues of chronic gastritis were collected at the same time,and miR-532-3p β-Expression of catenin and cyclin D1 and their relationship with clinicopathological features were analyzed.Pearson analyzed the relationship between miR-532-3p and β-catenin and cyclin D1 correlation,ROC curve was used to verify miR-532-3p β-The diagnostic efficacy of catenin and cyclin D1 in gastric cancer.Results:(1)Compared with gastric cancer tissues,miR-532-3p expression in gastritis tissues was increased,β-The expression of catenin and cyclin D1 was significantly decreased(P β-There was no significant difference in the relative expression of catenin and cyclin D1(P>0.05).(2)The expression of miR-532-3p in gastric cancer tissue was 0.89 ± 0.15,and the mean value was used as the baseline for classification.The patients with higher than the mean value were treated as the high expression group of miR-532-3p,and the patients with lower than the mean value were treated as the low expression group of miR-532-3p.The results of clinical characteristics of miR-532-3p and gastric cancer patients showed that there were significant differences between the lack of miR-532-3p expression and the tumor size>5cm.TNM staging increase,lymph node metastasis(P<0.05).and gender,age There was no significant difference in the degree of differentiation and infiltration(P>0.05).(3)Will β-catenin is grouped according to expression level,with high expression β-catenin was low expressed in 38 cases β-catenin was 42 cases,and cyclin D1 was grouped according to the expression level.35 cases had high expression of cyclin D1,and 45 cases had low expression of cyclin D1.β-The expression of catenin and cyclin D1 was significantly higher than TNM stage,lymph node metastasis and differentiation(P.(4)Comparison of miR-532-3p and β-The correlation analysis of catenin and Cyclin D1 shows that miR-532-3p and β-The levels of catenin and cyclin D1 showed negative correlation(P<0.05).(5)miR-532-3p、β-catenin and Cyclin D1 have similar diagnostic efficacy for gastric cancer alone.The combined detection of AUC area is 0.925,specificity is 93.30%.and sensitivity is 88.96%.indicating that the combined detection has high diagnostic efficacy for gastric cancer.Conclusion:(1)Compared with gastritis and adjacent tissues,miR-532-3p is lower expressed in gastric cancer.β-catenin and cyclin D1 were highly expressed in gastric cancer.(2)The loss of miR-532-3p expression was positively correlated with tumor size>5cm.TNM staging and lymph node metastasis of gastric cancer.β-The overexpression of catenin and cyclin D1 was positively correlated with the increase of TNM stage,lymph node metastasis and poor differentiation in gastric cancer patients.(3)miR-532-3p and β-The expression of catenin and cyclin D1 was negatively correlated.(4)miR-532-3p、β-The combined diagnosis of catenin and cyclin D1 has high diagnostic efficiency for gastric cancer.Part Ⅱ:Study on the mechanism of miR-532-3p regulatingβ-catenin/Cyclin D1 on migration and invasion of gastric cancer cellsObjective:To investigate the mechanism of miR-532-3p regulatingβ-catenin/Cyclin D1 on the migration and invasion of gastric cancer cells.Methods:Gastric cancer group(gastric cancer cell line without transfection),miR-532-3P-mimics NC group(gastric cancer cell line transfected with miR-532-3Pmimics NC),and miR-532-3p mimics group(gastric cancer cell line transfected with miR-532-3p mimics),miR-532-3p-inhibitor group(gastric cancer cell line transfected with miR-532-3p-inhibitor)and miR-532-3p-inhibitor NC group(gastric cancer cell line transfected with miR-532-3p-inhibitor NC).QPCR was used to detect the expression of miR-532-3p in cells of each group,and CCK-8 was used to detect the cell activity of gastric cancer cells at 24h,48h,72h and 96h.The apoptosis rate of each group was detected by flow cytometry.The migration distance of cells in each group was detected by scratch test,and the number of cell invasion was detected by Transwell chamber.Western blotting was used to detect the protein expressions of E-cadherin,N-cadherin,Vimentin,β-catenin and Cyclin D1 in each group.QPCR was used to detect the protein levels of β-catenin and Cyclin D1 in each group.Luciferase was used to verify the regulatory effect of miR-532-3p on β-catenin and Cyclin D1.Results:There was no significant difference in miR-532-3p,gastric cancer cell activity,migration,invasion,apoptosis and protein expression among gastric cancer group,miR-532-3p mics NC group and miR-532-3p inhibitor NC group(P>0.05);Compared with gastric cancer group and miR-532-3p mimics NC group,miR-532-3p expression,apoptosis rate and E-cadherin protein expression in gastric cancer cells of miR-532-3p mimics group increased,and gastric cancer cells’ 24h,48h,72h and 96h activity,invasion number,migration distance,N-cadherin,Vimentin and β-The expression of catenin and Cyclin D1 protein decreased(P<0.05).Compared with gastric cancer group and miR-532-3p inhibitor NC group,the expression of miR-532-3p,apoptosis,migration distance,N-cadherin,Vimentin and β-The expression of catenin and cyclin D1 increased(P<0.05);β-Catenin and Cyclin D1 were positively correlated with the proliferation,invasion and migration of gastric cancer cells(P;Overexpression of miR-532-3p can inhibit β-Catenin,cyclin D1 3 ’ UTR WT activity(P<0.05),butβ-The levels of catenin and cyclin D1 3’ UTR MUT had no effect(P>0.05).Compared with miR-532-3p mimics NC group,miR-532-3p mimics group and miR-532-3p mimics NC group+ β-Activity,migration distance,invasion number and βThe expression levels of catenin and cyclin D1 decreased,the apoptosis rate of gastric cancer cells increased,and miR-532-3p mimics NC+β-Catenin,cyclin D1 agonist group,gastric cancer cell activity,migration distance,invasion number and β The expression levels of catenin and cyclin D1 were increased,and the apoptosis rate of gastric cancer cells was decreased,with significant difference between groups(P<0.05):miR-532-3p mimics group and miR-532-3p mimics NC+β-Activity,migration distance.invasion number,apoptosis rate and β-There was no significant difference in the expression levels of catenin and cyclin D1(P>0.05);MiR-532-3p mimics NC group and miR-532-3p mimics+β-The levels of gastric cancer cells in cathin and cyclin D1 agonists groups were not statistically significant(P>0.05):miR-532-3p mimics+βActivity,migration distance,invasion number and β-The expression levels of catenin and cyclin D1 were lower than those in miR-532-3p mimics group and miR-532-3p mimics NC+β-Apoptosis rate of gastric cancer cells in cathin and cyclin D1 inhibitor group was higher than that in miR-532-3p mimics group and miR-532-3p mimics NC+β-Catenin and cyclin D1 inhibitor groups had statistically significant difference(P<0.05).Conclusion:(1)Transfection of miR-532-3p inhibitor can improve the activity,invasion and migration ability of gastric cancer cells,inhibit cell apoptosis,reduce the transformation of E-cadherin protein,and increase the expression levels of N-cadherin and Vimentin proteins.(2)miR-532-3p can inhibit the expression level of β-catenin/CyclinDl signaling pathway.(3)Transfection of miR-532-3p mimics can increase the level of E-cadherin protein transformation,reduce the expression levels of N-cadherin and Vimentin proteins,reduce the proliferation,invasion and migration of gastric cancer cells,and accelerate apoptosis..Cyclin D1 was positively correlated with gastric cancer cell proliferation,invasion and migration,and negatively correlated with gastric cancer apoptosis,indicating that the effect of miR-532-3p overexpression on gastric cancer cell activity may be related to the targeted inhibition of β-catenin/CyclinD1 expression. |
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