miR-125a-5p通过调控IGSF3表达抑制食管鳞状细胞癌迁移侵袭能力

发布时间：2018-12-30 15:45

【摘要】：背景越来越多的证据表明,miR-125a-5p可以作为多种癌症的肿瘤抑制基因;然而,其作用于食管鳞癌的机制知之甚少。目的(1)筛选食管鳞癌患者中异常表达的miRNA及功能验证;(2)探讨miRNA潜在靶基因。方法(1)应用qRT-PCR检测miRNA在ESCC中的表达水平;(2)应用体内体外功能实验探究miRNA对ESCC生物学行为的影响;(3)应用双荧光素酶实验对miRNA在ESCC中作用机制进行初步研究。结果我们利用芯片检测技术发现miR-125a-5p在食管鳞癌组织中的相对表达程度低于相应的癌旁正常组织,并通过血清样本对这一成果进行了验证。另外发现,miR-125a-5p与T分期及临床分期相关。进一步的体内体外功能实验证实,构建过表达miR-125a-5p的TE-1和ECA109人食管鳞癌细胞系可抑制肿瘤细胞的增殖、G1/S期过渡、迁移和侵袭,同时也可促进肿瘤细胞的凋亡。更重要的是,IGSF3是免疫球蛋白超家族(IGSF)的一员,研究认为它可以通过多种途径参与肿瘤迁移和侵袭能力的调控。利用双荧光素酶实验证明IGSF3是miR-125a-5p的直接作用靶基因;通过将无3?-UTR区域的shRNA-IGSF3和miR-125a-5p mimics共转染TE-1和ECA109细胞系进行功能实验,发现IGSF3低表达能够增强miR-125a-5p mimics对食管鳞癌细胞系迁移及侵袭能力抑制作用。结论我们首次发现了miR-125a-5p和IGSF3在食管鳞癌中的相互作用关系,为miR-125a-5p作为食管鳞癌诊断的肿瘤志物及通过IGSF3这一靶基因指导食管鳞癌的治疗提供一个新思路。
[Abstract]:Background there is growing evidence that miR-125a-5p can be used as a tumor suppressor gene in many cancers; however, little is known about the mechanism of its role in esophageal squamous cell carcinoma. Objective (1) to screen the abnormal expression of miRNA and function in esophageal squamous cell carcinoma patients and (2) to explore the potential target gene of miRNA. Methods (1) the expression of miRNA in ESCC was detected by qRT-PCR; (2) the effect of miRNA on the biological behavior of ESCC was investigated by in vivo and in vitro functional experiments; (3) the mechanism of miRNA in ESCC was studied by using double luciferase assay. Results We found that the relative expression of miR-125a-5p in esophageal squamous cell carcinoma tissues was lower than that in adjacent normal tissues by using microarray technique, and the results were verified by serum samples. In addition, miR-125a-5p was associated with T stage and clinical stage. Further in vitro and in vivo functional experiments confirmed that the construction of TE-1 and ECA109 human esophageal squamous cell carcinoma cell lines expressing miR-125a-5p could inhibit the proliferation of tumor cells, the G 1 / S phase transition, migration and invasion, but also promote the apoptosis of tumor cells. More importantly, IGSF3 is a member of immunoglobulin superfamily (IGSF). Double luciferase assay showed that IGSF3 was a direct target gene of miR-125a-5p. By co-transfection of shRNA-IGSF3 and miR-125a-5p mimics without 3?-UTR region into TE-1 and ECA109 cell lines, it was found that the low expression of IGSF3 could enhance the inhibitory effect of miR-125a-5p mimics on the migration and invasion of esophageal squamous cell carcinoma cell lines. Conclusion the interaction between miR-125a-5p and IGSF3 in esophageal squamous cell carcinoma was first discovered, which provides a new idea for miR-125a-5p as a tumor marker for diagnosis of esophageal squamous cell carcinoma and for guiding the treatment of esophageal squamous cell carcinoma by IGSF3.
【学位授予单位】：济南大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.1

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