中图分类号R739.65 学校代码10634
连续缠绕玻璃钢夹砂管UDC610 密级
授予单位川北医学院
H19/miR-675-5p与食管鳞状细胞癌上皮间质转化的相关性研究The Study on the Expression and Function of H19/miR-675-5p in Epithelial-mesenchymal Transition for Esophageal Squamous Cell Carcinoma
作者张倩包装箱制作
导师文世民教授
培养院系临床医学系
授位类别学术学位
授位级别硕士
学科门类医学
小麦玉米联合收割机
研究方向食管癌
在线测厚
年级2016级
学号22016044
二○一九年五月
H19/miR-675-5p与食管鳞状细胞癌上皮间质转化的相
关性研究
摘 要
目的：长链非编码RNA（long-nocoding RNA,lncRNA）H19一直以来被认为与肿瘤的发生发展过程密切相关，与其产物miR-675-5p在多种实体肿瘤中异常表达，有学者发现H19/miR-675-5p共表达升高是结肠癌细胞发生上皮间质转化（Epithelial-mesenchymal transition,EMT）的重要机制。但是由于肿瘤的异质性，同一种基因在不同肿瘤中可能发挥截然不同的作用，因此H19/miR-675-5p在食管鳞癌（ esophageal squamous cell carcinoma,ESCC）中的作用及是否与ESCC细胞EMT过程相关尚不明确。本研究旨在探讨H19/miR-675-5p共表达水平与ESCC患者临床分期的关系，利用siRNA基因干扰技术明确H19/miR-675-5p是否与ESCC细胞株EMT过程相关。 方法：
1 通过RT-PCR技术检测H19/miR-675-5p在不同ESCC患者肿瘤组织及配对正常食管组织、ESCC细胞株及正常食管上皮细胞株表达水平，分析H19/miR-675-5p共表达与食管鳞癌患者TNM分期的相关性；
2 利用质粒瞬转ESCC细胞株ECA109，siRNA基因干扰技术，下调H19/miR-675-5p表达，分析下调表达后EMT相关蛋白表达及细胞株迁移及侵袭能力的变化。
结果：
1 H19/miR-675-5p在ESCC肿瘤组织及细胞株中表达水平均明显上调，且二者高表达水平与患者T、N分期晚具有相关性，H19/miR-675-5p
表达水平升高在淋巴结转移阳性的患者中尤其明显；
2 下调H19/miR-675-5p表达后，ECA109细胞株迁移及侵袭能力显著下降，且上皮特性蛋白表达上升，间质特性蛋白表达下降。
结论：H19/miR-675-5p共表达升高能促进食管鳞癌细胞发生EMT，从而增强肿瘤细胞侵袭及转移能力，因此H19/miR-675-5p共表达水平或许可作为ESCC患者预测预后的分子标志物，指导其共表达显著上升的患者选择更加积极的方案。
关键词：H19/miR-675-5p；上皮间质转化；食管鳞癌；预后
The study on the expression and fuction of
H19/miR-675-5p in Epithelial-mesenchymal transition for esophageal squamous cell carcinoma
AbstractDAPD001
Objective: Long-noncoding RNA (lncRNA) H19 has long been thought to be closely related to the development and progression of tumors. Its product miR-675-5p is abnormally expressed in various solid tumors. Some scholars have found that elevated expression of H19/miR-675-5p is an important mechanism for epithelial-mesenchymal transition (EMT) in colon cancer cells. Due to the heterogeneity of tumor cells, the same gene may play a distinct role in different tumors. Therefore, the role of H19/miR-675-5p in esophageal squamous cell carcinoma (ESCC) and whether it is related to the relevance of cellular EMT processes is not known. This study was to investigate the rel
ationship between the co-expression level of H19/miR-675-5p and the clinical stage of ESCC patients. The siRNA gene interference technology was used to determine whether H19/miR-675-5p is related to the EMT process in ESCC cell line.
Methods: 1.RT-PCR was used to compare the expression of H19/miR-675-5p in different ESCC patients and the normal esophageal tissue, ESCC cell line and normal esophageal epithelial cell line. And then to analysethe correlationof expression of H19/miR-675-5p with TNM staging in esophageal squamous cell carcinomapatients;
2. Used siRNA gene interference technology to knock downregulating expressionof H19/miR-675-5p. Then analysed down-regulated expression of EMT-related gene and detected the插板闸门
