摘要
目的探讨miR-223作用于STAT3对胃癌细胞增殖与迁移的影响,并揭示其分子作用机制。方法采用实时荧光定量PCR(q-PCR)法检测miR-223在胃癌组织与癌旁组织及正常胃黏膜细胞(GSE-1)与胃癌细胞系MGC-803、SGC-7901和BCG-823中的表达情况;分别将miR-223模拟物(miR-223 mimics)、anti-miR-223 mimics和NC mimics转染入SGC-7901细胞,分析miR-223对SGC-7901增殖的调控作用,采用Targetscan和miRdata等生物信息学预测软件对miR-223的作用靶点进行预测,采用双荧光素酶报告基因系统验证miR-223与STAT3的作用关系,采用q-PCR法及Western Blot法分别检测miR-223对胃癌细胞SGC-7901中STAT3 mRNA及蛋白表达的影响。结果与癌旁组织比较,胃癌组织中miR-223的表达水平明显降低,胃癌细胞系MGC-803、SGC-7901和BCG-823中miR-223的表达水平较GSE-1细胞明显降低,相比于NC mimics,转染miR-223 mimics之后SGC-7901细胞的增殖显著抑制,双荧光素酶报告基因系统检测结果显示,miR-223可直接调控STAT3的转录活性,且miR-223可显著抑制SGC-7901细胞中STAT3蛋白的表达。结论miR-223可靶向作用于STAT3从而调控胃癌细胞的增殖与迁移。
Objective To investigate the effect of miR-223 on the proliferation and migration of gastric cancer cell line,and to reveal its possible molecular mechanism.Methods The expression of miR-223 in gastric cancer tissues,normal gastric mucosa cells(GSE-1)and gastric cancer cell lines MGC-803,SGC-7901 and BCG-823 were detected by real time fluorescent quantitative PCR(q-PCR).The miR-223 mimics,anti-miR-223 mimics and NC mimics were transfected into SGC-7901 cells,respectively,and the effects of miR-223 on the proliferation of SGC-7901 were analyzed.Bioinformatics prediction software such as Targetscan and microarray data were used.The target of miR-223 was predicted,and the relationship between miR-223 and STAT3 was verified by double luciferase reporter gene system.The effects of miR-223 on the expression of STAT3 in gastric cancer cell SGC-7901 were detected by q-PCR and Western Blot,respectively.Results Compared with the adjacent tissues,the expression level of miR-223 in gastric cancer tissues was significantly lower.The expression level of miR-223 in gastric cancer cell lines MGC-803,SGC-7901 and BCG-823 was significantly lower than that in GSE-1 cells.Compared with NC mimics,the proliferation of SGC-7901 cells was significantly inhibited after miR-223 MICs transfection.The results of double luciferase reporter gene system showed that miR-223 could directly regulate STAT3.The transcriptional activity,and the expression of STAT3 in SGC-7901 cells were significantly inhibited by miR-223.Conclusion miR-223 could target STAT3 to regulate the proliferation of gastric cancer cells.
作者
解小霞
孙晓静
苏文炀
杨柳
李力
王薇
李婷
田永强
XIE Xiaoxia;SUN Xiaojing;SU Wenyang;YANG Liu;LI Li;WANG Wei;LI Ting;TIAN Yongqiang(Wuhan Hospital of Traditional Chinese Medicine,Wuhan 430014,China;Hubei Hospital of Traditional Chinese Medicine,Wuhan 430061,China)
出处
《西北药学杂志》
CAS
2020年第4期526-530,共5页
Northwest Pharmaceutical Journal
基金
国家自然科学基金项目(编号:81803676)
湘潭市科技局项目(编号:ZJ20171001)。