miR-216b-5p在肾癌中的表达及其功能研究

发布时间：2018-02-06 07:24

本文关键词： 肾癌 miR-216b-5p 表达功能　出处：《安徽医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:肾细胞癌(renal cell carcinoma,RCC)简称肾癌,是泌尿生殖系统中最常见的恶性肿瘤之一,也是肾脏实质中最常见的恶性肿瘤,其恶性程度较高。由于肾癌患者早期症状和体征往往不明显,缺乏特异性的临床表现,因此早期诊断较为困难,以致约三分之一的肾癌患者在明确诊断时肿瘤已经发生了远处脏器转移。由于肾癌对放疗和普通化疗不敏感,手术切除病灶成为有效的治疗方法,但是仍有20-30%的肾癌患者在接受手术治疗后出现了肿瘤复发。因此,研究肾癌发生发展的作用机制并寻找肾癌早期诊断、疗效评价、预后评估的分子标志物就显得尤为重要。微小RNA(micro RNA或者miRNA)是一类具有大约21-23个核苷酸长度的短链非编码RNA分子,它主要通过与靶基因的m RNA 3’端非编码区(3'-UTR区)一些特异性序列结合,从而降解靶基因m RNA或者抑制其翻译,在转录后水平起到调控基因表达的作用。目前的研究表明,miRNA在包括肾癌在内的多种肿瘤组织中异常表达,并参与细胞增殖、侵袭、迁移和凋亡等生命过程之中。在此我们以miR-216b-5p和肾癌作为研究对象,研究miR-216b-5p在肾癌发生发展过程中的功能及相关致癌机制,为后续开展miRNA在肾癌诊断和治疗中的研究提供一些理论基础。方法:应用qRT-PCR方法检测41例肾癌组织和配对癌旁正常组织中miR-216b-5p的表达水平,并使用统计学软件分析miR-216b-5p在肾癌组织中的表达水平与患者临床病理特征之间的相关性。然后分别通过CCK-8增殖实验、细胞划痕实验和流式细胞术分别检测miR-216b-5p对肾癌细胞增殖、迁移和凋亡的影响。最后通过生物信息学方法预测miR-216b-5p的潜在靶基因。结果:与配对癌旁正常组织相比,miR-216b-5p在肾癌组织中的表达水平明显降低(P0.05),而且其在肾癌组织中的表达水平与患者性别、年龄、病理类型、TNM分期、临床AJCC分期之间无明显相关性(P0.05)。通过转染miR-216b-5pmimics上调肾癌细胞中miR-216b-5p的表达水平后,与对照组相比,细胞增殖和迁移能力受到抑制,同时促进了细胞凋亡(P0.05)。结论:miR-216b-5p在肾癌组织中表达下调,并通过抑制肾癌细胞增殖和迁移,促进肾癌细胞凋亡,从而发挥抑癌基因作用。KRAS可能是miR-216b-5p的其中一个靶基因。
[Abstract]:Objective: renal cell carcinoma (RCC) is one of the most common malignant tumors in the genitourinary system. It is also the most common malignant tumor in renal parenchyma, its malignant degree is high. Because the early symptoms and signs of renal carcinoma patients are often not obvious, lack of specific clinical manifestations, so the early diagnosis is more difficult. As a result, about 1/3 of RCC patients had distant organ metastasis at the time of definite diagnosis. Because RCC is insensitive to radiotherapy and chemotherapy, surgical resection of tumor has become an effective treatment method. However, there are still 20-30% of patients with renal cell carcinoma have recurrence after surgery. Therefore, to study the mechanism of the development of renal cell carcinoma and to search for early diagnosis of renal cell carcinoma, evaluation of curative effect. Molecular markers for prognostic evaluation are particularly important. Small RNA(micro RNA or miRNAs. It is a class of short chain noncoding RNA molecules with about 21-23 nucleotide lengths. It degrades the target gene m RNA or inhibits its translation by binding to some specific sequences of the target gene in the 3'-terminal non-coding region of the target gene. At the post-transcriptional level, it plays a role in regulating gene expression. Current studies have shown that miRNA is abnormal expressed in various tumor tissues, including renal cell carcinoma, and participates in cell proliferation and invasion. In the process of migration and apoptosis, we focus on miR-216b-5p and renal cell carcinoma (RCC). To study the function of miR-216b-5p in the development of renal cell carcinoma and its carcinogenic mechanism. To provide a theoretical basis for the further study of miRNA in the diagnosis and treatment of renal cell carcinoma. Methods:. QRT-PCR method was used to detect the expression of miR-216b-5p in 41 cases of renal cell carcinoma and adjacent normal tissues. Statistical software was used to analyze the correlation between the expression level of miR-216b-5p and the clinicopathological characteristics of RCC. Then CCK-8 proliferation test was used. Cell scratch assay and flow cytometry were used to detect the proliferation of RCC cells by miR-216b-5p. The effects of migration and apoptosis. Finally, the potential target genes of miR-216b-5p were predicted by bioinformatics. Results: compared with matched adjacent normal tissues. The expression level of miR-216b-5p in RCC was significantly lower than that in RCC, and its expression level was correlated with sex, age and pathological type of RCC. There was no significant correlation between clinical AJCC staging and P0.05.After transfection of miR-216b-5pmimics, the expression of miR-216b-5p in RCC cells was up-regulated. Compared with the control group, cell proliferation and migration were inhibited and apoptosis was promoted by P0.05.Conclusion the expression of micromiR-216b-5p is down-regulated in renal cell carcinoma. KRAs may be one of the target genes of miR-216b-5p by inhibiting the proliferation and migration of RCC cells and promoting the apoptosis of RCC cells.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.11

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