食品工业科技2024，Vol.45Issue(14)：335-343,9.DOI:10.13386/j.issn1002-0306.2023080155
灰树花多糖通过LncRNA HULC/miR-186-5p轴调控胆囊癌细胞增殖、迁移和侵袭
Grifola frondose Polysaccharide Regulates Proliferation,Migration and Invasion of Gallbladder Cancer Cells Through LncRNA HULC/miR-186-5p Axis
摘要
Abstract
Objective:To analyze the effects of Grifola frondose polysaccharide(GFP)on proliferation,migration and invasion of gallbladder carcinoma cells and its mechanism.Methods:Logarithmic phase GBC-SD cells randomized:Control group,GFP group(0.5,1,2 μg/mL GFP),shNC group,shHULC group,miR-NC group,miR-186-5p mimics group,GFP+pcDNA group,GFP+pcDNA-HULC group.Cell inhibition rate was detected by CCK-8 method.The migration and invasion of GBC-SD cells were detected by Transwell cell assay.The protein levels of Cyclin D1,P21,MMP-2 and MM-9 were detected by Western Blot.LncRNA HULC and miR-186-5p levels were detected by qRT-PCR.Starbase online software predicted the binding sites of HULC and miR-186-5p,dual luciferase reporting assay was used to detect the targeting relationship between LncRNA HULC and miR-186-5p.The effect of GFP on the endogenetic growth of GBC-SD cells was detected by tumor transplantation in nude mice.Results:0.25～8 μg/mL GFP had definite inhibitory effect on GBC-SD cells of gallbladder carcinoma.Compared with the control group,0.5,1 and 2 μg/mL of GFP(extremely)significant(P＜0.05,P＜0.01)inhibited the migration and invasion of GBC-SD cells,and(extremely)significant(P＜0.05,P＜0.01)decreased the levels of Cyclin D1,P21,MMP-2 and MM-9,extremely significant increased the levels of P21(P＜0.05,P＜0.01).The expression of LncRNA HULC was extremely significant down-regulated and miR-186-5p was extremely significant up-regulated(P＜0.05,P＜0.0l).Starbase online software predicted that there was a binding site between HULC and miR-186-5p,and compared with miR-NC group,the luciferase activity of HULC-WT in miR-186-5p mimics group was extremely significant(P＜0.01)decreased.There was no significant difference in the luciferase activity of HULC-MUT(P＞0.05).Compared with shNC group,the relative expression level of HULC in shHULC group was extremely significant(P＜0.01)decreased,the relative expression level of miR-186-5p was extremely significant(P＜0.01)increased,the cell survival rate,migration number and invasion number were significantly(P＜0.05)decreased,respectively,the levels of MMP-2,MMP-2 and MM-9 were extremely significant decreased(P＜0.01).Compared with GFP+pcDNA group,the inhibition rate of GFP+pcDN-HULC group significantly decreased(P＜0.05),and the number of migration and invasion was significantly increased(P＜0.01),the CylinDl,MMP-2 and MM-9 levels were significantly increased(P＜0.05,P＜0.01).Compared with GFP+pcDNA administration group,tumor volume and mass in GFP+pcDN-HULC administration group were significantly(P＜0.05)increased,and miR-186-5p levels were extremely significantly decreased(P＜0.01).Conclusion:GFP could inhibit proliferation,invasion and migration of gallbladder cancer cells,and its mechanism was related to the regulation of LncRNA HULC/miR-186-5p.关键词
灰树花多糖/LncRNA HULC/miR-186-5p/胆囊癌/增殖/迁移/侵袭Key words
Grifola frondose polysaccharide/LncRNA HULC/miR-186-5p/gallbladder cancer/proliferation/migration/invasion分类
医药卫生引用本文复制引用
于静,段子朋,徐松涛..灰树花多糖通过LncRNA HULC/miR-186-5p轴调控胆囊癌细胞增殖、迁移和侵袭[J].食品工业科技,2024,45(14):335-343,9.基金项目
河南省二0二二年科技发展计划(222102310443) （222102310443）
漯河医学高等专科学校2023年度科技创新项目(2023ZD04). （2023ZD04）
