喉鳞状细胞癌特征性microRNA表达谱的研究

发布时间：2018-10-05 11:58

【摘要】： 占人类基因1%的microRNA (miRNA)几乎调节着至少30%的mRNA,它们通过调控相关靶基因mRNA的表达水平影响肿瘤细胞的表型。喉癌是最常见的头颈部恶性肿瘤,也是仅次于肺癌的呼吸道第二高发癌。目前,尚少见有喉癌的miRNA特征性表达谱或与其生物学行为相关的差异表达miRNA的研究报道。 本项研究采用基于基因芯片技术的miRNA表达谱的分析方法,分别以福尔马林固定石蜡包埋(formalin-fixed and paraffin-embedded, FFPE)组织和新鲜冻存组织为研究材料,以获得喉鳞状细胞癌特征性的miRNA表达谱型；进而筛选出与喉鳞癌生物学特征密切相关的差异表达miRNA。同时比较从FFPE和新鲜冻存两种组织材料中所获得的喉鳞癌miRNA表达谱的差异,评价FFPE组织miRNA表达谱分析的有效性和可行性。这项研究的远期目标为,从上述“分子变化谱型”中筛选出具有鉴别意义的miRNA,对喉癌发生与发展、淋巴结转移或复发风险进行评估预测,以辅助诊断和指导治疗。 首先,应用Agilent公司的人类miRNA芯片,针对24例喉鳞癌FFPE组织样品以及8例新鲜冻存的喉鳞癌患者配对癌组织和癌旁组织样品的总RNA进行了miRNA表达谱检测。继而,采用基因芯片分析软件GeneSpring和统计学软件R project分析了上述miRNA芯片的实验数据。结果显示,从24例喉鳞癌FFPE织标本中鉴定到319个miRNA,其中96个miRNA在全部24例中均有表达；从8例喉鳞癌新鲜冻存癌组织标本中鉴定到342个miRNA,其中161个miRNA在全部8例中均有表达；从而初步获得喉鳞癌的miRNA表达谱。在FFPE组织中检测到的319个miRNA信号中有233个也能在新鲜冻存喉癌组织中检测到；在FFPE组织中检测到的79个与淋巴结转移相关(p0.05)的miRNA中有16个也在新鲜冻存喉癌组织中发现的与淋巴结转移相关的52个miRNA中出现,这16个miRNA包括miR-17、miR-130b、miR-183、miR-30b、miR-20a、miR-96、miR-188-5p等；从而确认了上述FFPE组织miRNA表达谱分析的有效性。进而采用检错率FDR0.05进行校正,以筛选FFPE组织样品中显著差异表达的miRNA,获得5个与喉鳞癌淋巴转移密切相关的差异表达miRNA:miR-23a*、miR-28-5p、miR-15a、miR-16和miR-425。 本项研究表明,具有不同生物学行为的喉鳞癌之间存在miRNA的差异表达,miR-23a*、miR-28-5p、miR-15a、miR-16和miR-425与喉鳞癌淋巴转移密切相关,有望通过进一步扩大样本的深入研究建立起能够起到鉴别或评估预测作用的分子模型。而医院病理科存档的FFPE组织可以提供符合基因芯片分析质量要求的miRNA,是研究miRNA的重要样品资源。
[Abstract]:MicroRNA (miRNA), which accounts for 1% of human genes, almost regulates at least 30% of mRNA,. It affects the phenotype of tumor cells by regulating the expression level of target gene mRNA. Laryngeal cancer is the most common malignant tumor of the head and neck and the second most common cancer of the respiratory tract after lung cancer. At present, there are few studies on miRNA characteristic expression profile or differential expression of miRNA related to biological behavior of laryngeal carcinoma. In this study, the miRNA expression profile of laryngeal squamous cell carcinoma was obtained by using formalin fixed paraffin embedded (formalin-fixed and paraffin-embedded, FFPE) tissue and fresh frozen tissue as materials. Furthermore, the differential expression of miRNA. closely related to the biological characteristics of laryngeal squamous cell carcinoma was screened out. At the same time, the differences of miRNA expression profiles of laryngeal squamous cell carcinoma (LSCC) obtained from FFPE and fresh frozen tissue materials were compared to evaluate the validity and feasibility of miRNA expression profile analysis in FFPE tissues. The long-term objective of this study is to screen out the differential miRNA, from the above "molecular change patterns" to evaluate and predict the risk of laryngeal cancer occurrence and development, lymph node metastasis or recurrence, in order to assist in diagnosis and guidance of treatment. Firstly, the total RNA expression profiles of 24 laryngeal squamous cell carcinoma (LSCC) FFPE tissues and 8 fresh frozen laryngeal squamous cell carcinoma (LSCC) samples were detected by Agilent human miRNA microarray in matched tissues and adjacent tissues of laryngeal squamous cell carcinoma (LSCC). Then, GeneSpring and R project were used to analyze the experimental data of these miRNA chips. The results showed that 319 miRNA, samples were identified from 24 cases of laryngeal squamous cell carcinoma (LSCC), 96 miRNA were detected in all 24 cases, 342 miRNA, were identified from 8 samples of fresh frozen tissue of laryngeal squamous cell carcinoma (LSCC), and 161 miRNA were detected in all 8 cases. Thus the miRNA expression profile of laryngeal squamous cell carcinoma was preliminarily obtained. 233 of the 319 miRNA signals detected in FFPE tissues could also be detected in fresh frozen laryngeal cancer tissues. Sixteen of 79 miRNA associated with lymph node metastasis (p0.05) detected in FFPE tissues were also found in 52 miRNA related to lymph node metastasis in fresh frozen laryngeal carcinoma tissues, including miR-17,miR-130b,miR-183,miR-30b,miR-20a,miR-96,miR-188-5p. Therefore, the validity of miRNA expression profile analysis of FFPE tissue was confirmed. Then the error detection rate (FDR0.05) was used to correct for the screening of significantly differentially expressed miRNA, in FFPE tissue samples. Five differentially expressed miRNA:miR-23a*,miR-28-5p,miR-15a,miR-16 and miR-425. were obtained which were closely related to lymphatic metastasis of laryngeal squamous cell carcinoma (LSCC). This study shows that the differential expression of miRNA in laryngeal squamous cell carcinoma with different biological behaviors is closely related to lymphatic metastasis of laryngeal squamous cell carcinoma. It is expected that a molecular model which can identify or evaluate the prediction can be established by further expanding the study of the sample. The FFPE tissue archived in hospital pathology department can provide miRNA, which meets the quality requirement of gene chip analysis. It is an important sample resource to study miRNA.
【学位授予单位】：中国协和医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R739.6

【引证文献】

相关博士学位论文 前1条

1 陈爱荣;1、叶酸代谢酶相关亚甲基四氢叶酸还原酶基因多态性与2型糖尿病及代谢综合征易感性的关联研究 2、膀胱癌石蜡包埋组织microRNA检测及临床意义的初步探讨[D];兰州大学;2010年

，

本文编号：2253292