miR-1184对幽门螺杆菌感染胃癌细胞生物学行为的影响OA
Effect of miR-1184 on biological behavior of gastric cancer cells infected by Helicobacter pylori
目的:探讨微小RNA-1184(miR-1184)对幽门螺杆菌(Helicobacter pylori,Hp)感染MGC-803 细胞增殖、迁移和炎症的影响.方法:应用实时荧光定量PCR检测miR-1184 在Hp感染MGC-803 细胞中的表达情况,将MGC-803 细胞分为模拟物对照组(miR-NC组)、miR-1184 模拟物组(miR-1184 组)、Hp感染组(miR-NC+Hp组)、miR-1184 模拟物联合Hp感染组(miR-1184+Hp组)、抑制剂对照组(In-NC组)和 miR-1184 抑制剂组(anti-miR-1184组);采用CCK-8实验、平板克隆实验、流式细胞术、Transwell实验、划痕愈合实验和蛋白质印迹法检测miR-1184 和Hp感染对胃癌细胞增殖、细胞周期、迁移和相关蛋白表达水平的影响.结果:与未感染组相比,Hp感染MGC-803细胞后引起细胞病变效应,miR-1184 表达水平降低(P<0.01).与miR-NC组相比,miR-1184 组MGC-803 细胞活性降低,平板克隆形成能力降低,G1 期细胞增多、S期和G2 期减少,划痕愈合能力减弱,可上调E-钙黏蛋白表达水平、降低N-钙黏蛋白和波形蛋白表达水平,抑制上皮间充质转化(EMT)进程(P<0.05 或P<0.01).与miR-NC+Hp组相比,miR-1184+Hp组抑制MGC-803 细胞增殖、细胞周期、迁移能力和EMT进程(P<0.05 或P<0.01).与miR-NC+Hp组相比,miR-1184+Hp组降低MGC-803 细胞中TNF-α、IL-1β、IL-6、IL-8 mRNA表达水平,抑制NF-κB p-p65/NF-κB p65 蛋白表达比例(P<0.05).抑制miR-1184 则促进MGC-803 细胞的增殖、周期、迁移和EMT进程(P<0.05或P<0.01).结论:miR-1184 在Hp感染胃癌细胞中表达下降,高表达miR-1184 可逆转Hp感染对胃癌细胞的增殖、迁移能力和炎症反应的影响.
Objective:To investigate the expression of microRNA-1184(miR-1184)in MGC-803 cells infected by Helicobacter pylori(Hp),and its effects on the proliferation,metastasis and inflammation of MGC-803 cells infected by Hp.Methods:Fluorescence quantitative PCR was used to detect the expression of miR-1184 in Hp-infected MGC-803 cells.MGC-803 cells were divided into control group(miR-NC group),miR-1184 mimics group(miR-1184 group),Hp infection group(miR-NC+Hp group),miR-1184 mimics combined with Hp infection group(miR-1184+Hp group),inhibitor control group(In-NC group)and miR-1184 inhibitor group(anti-miR-1184 group).The effects of miR-1184 and Hp infection on the proliferation,cell cycle,metastasis and expression levels of related proteins of gastric cancer cells were detected by CCK-8 assay,colony formation assay,cell cycle assay,Transwell assay,wound healing assay and Western blotting assay.Results:Compared to the uninfected group,Hp infected MGC-803 cells produced cytopathic effect,and decreased the expression of miR-1184(P<0.01).After transfection of miR-1184 mimics in MGC-803 cells,miR-1184 expression increased,cell activity decreased,colony formation ability decreased,G1 phase cells increased,S phase and G2 phase decreased,and wound healing ability decreased.In addition,miR-1184 mimics up-regulated the expression level of E-cadherin,decreased the expression level of N-cadherin and Vimentin,inhibited the process of epithelial-mesenchymal transformation(EMT)(P<0.05 or P<0.01).Compared with miR-NC+Hp group,miR-1184+Hp group inhibited MGC-803 cell proliferation,cell cycle,migration ability and EMT effects(P<0.05 or P<0.01).Compared with miR-NC+Hp group,miR-1184+Hp group decreased the mRNA expression of TNF-α,IL-1β,IL-6 and IL-8 in MGC-803 cells,and inhibited the proportion of NF-κB p-p65/NF-κB p65 protein expression(P<0.05).Furthermore,miR-1184 inhibitors promoted the proliferation,cell cycle,cell transfer and EMT process of MGC-803 cells(P<0.05 or P<0.01).Conclusion:The expression of miR-1184 is decreased in Hp-infected gastric cancer cells,and the high expression of miR-1184 reversed the effects of Hp infection on the proliferation,migration and inflammatory response of gastric cancer cells.
蒋颜材;姚镇东;王秀平;刘云;邵世和;纪筠
江苏大学附属宜兴医院普外科,江苏宜兴 214206江苏大学附属宜兴医院消化内科,江苏宜兴 214206江苏大学附属昆山医院检验科,江苏 昆山 215301江苏大学附属宜兴医院消化内科,江苏宜兴 214206江苏大学附属宜兴医院消化内科,江苏宜兴 214206江苏大学附属宜兴医院内镜中心,江苏 宜兴 214206
临床医学
miR-1184幽门螺杆菌胃癌细胞周期迁移NF-κB
miR-1184Helicobacter pylorigastric cancercell cyclemigrationNF-κB
《江苏大学学报（医学版）》 2025 (2)
125-132,8
国家资助博士后研究人员计划(GZC20230992)江苏省卓越博士后计划(2023ZB180)无锡市卫生健康委员会青年基金资助项目(Q202062)宜兴市卫生健康委员会重大项目(YXKY202209)江苏大学医教协同重点项目(JDY2023013)
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