miR-718和EGR3参与调控肝癌细胞系的恶性表型的研（英文）

发布时间：2018-01-30 01:24

本文关键词： miR- 小非编码RNA 早期生长反应蛋白肝癌表型　出处：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》2017年01期 　论文类型：期刊论文

【摘要】：目的:研究mi R-718和早期生长反应蛋白3(EGR3)在肝癌细胞系Hep G2及SMMC-7721中对细胞恶性行为的作用。创新点:首次在肝癌细胞系Hep G2和SMMC-7721中发现miR-718作为抑癌基因及EGR3作为促癌基因起到调控肿瘤恶性行为的作用。方法:采用聚合酶链反应(PCR)方法构建mi R-718和EGR3的过表达及敲降质粒,并采用实时定量PCR(q RT-PCR)方法验证质粒有效性;采用MTT法和克隆形成实验检测肝癌细胞系HepG 2和SMMC-7721的生长增殖能力;采用Transwell迁移侵袭实验检测肝癌细胞系Hep G2和SMMC-7721的迁移和侵袭能力;采用增强型绿色荧光蛋白(EGFP)荧光报告载体实验验证miR-718的靶基因;采用qR T-PCR和蛋白质印迹(Western blot)检测相关基因表达水平的变化。结论:mi R-718在肝癌细胞系HepG 2和SMMC-7721中起到抑癌基因的作用;EGR3在肝癌细胞系Hep G2和SMMC-7721中起到促癌基因的作用;miR-718是通过靶定EGR3 m RNA 3'UTR下调EGR3的表达起到抑癌基因的作用。
[Abstract]:Objective: to study the expression of mi R-718 and early growth response protein 3 (EGR3). Effect of hepatoma cell line Hep G2 and SMMC-7721 on malignant behavior of hepatoma cell line. Innovation: for the first time in hepatocellular carcinoma cell line Hep. MiR-718 as tumor suppressor gene and EGR3 as promoter gene were found in G2 and SMMC-7721. Methods: polymerase chain reaction (PCR) was used to regulate the malignant behavior of tumor. The overexpression and knockdown plasmid of mi R-718 and EGR3 were constructed by PCR. The validity of plasmid was verified by real-time quantitative PCR(q RT-PCR method. The growth and proliferation of hepatoma cell line HepG 2 and SMMC-7721 were detected by MTT assay and clone forming assay. The migration and invasion ability of hepatoma cell line Hep G2 and SMMC-7721 were detected by Transwell migration and invasion assay. The enhanced green fluorescent protein (EGFP) fluorescent report vector was used to verify the target gene of miR-718. QR T-PCR and Western blot were used. Conclusion: the cell line HepG 2 and SMMC-7721 play an important role in inhibiting oncogene expression. EGR3 plays the role of oncogene promoter in Hep G2 and SMMC-7721 cell lines. MiR-718 plays a role in tumor suppressor gene by targeting EGR3 m RNA 3 UTR to down-regulate the expression of EGR3.
【作者单位】： Clinical
【基金】：Project supported by the Science and Technology Project of Higher Education of Shandong Province(No.J12LK07),China
【分类号】：R735.7
【正文快照】： 1 IntroductionHepatocellular carcinoma(HCC)is one of themost prevalent and aggressive human malignancies worldwide,and it has a very poor prognosis(Walleret al.,2015).Men are more susceptible to HCC thanwomen and it is a leading cause of cancer-relatedd

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