食管鳞癌中微小rnalet-7的作用及其对高迁移率蛋白a2的影响-effect of small rna let - 7 on high mobility protein a2 in esophageal squamous cell carcinoma.docxVIP

新疆 新疆医科大学硕士学位论文 - - PAGE 1 - 食管鳞癌中微小 RNA let-7 的作用及其对高迁移率蛋白 A2 的影响 研究生：刘清 导师：卢晓梅 教授 摘 要 目的：探讨食管鳞癌中微小RNA(miRNA)let-7的作用,及其对高迁移率蛋白A2(High mobility group A2 protein，HMGA2)的调控影响。 方法：在细胞水平，应用RNA干扰 (RNAi)技术，将let-7及其抑制剂分别转染人食管鳞癌细胞系Eca109，利用实时荧光定 量聚合酶链式反应(Quantitative real-time PCR，qRT-PCR)方法分别检测let-7及 HMGA2 mRNA的表达情况，蛋白印迹(Western blot)检测HMGA2蛋白的表达变化； 在组织水平，应用qRT-PCR方法检测45例食管鳞癌及其对应的癌旁食管组织中let-7 和HMGA2 mRNA的表达情况，同时基于免疫组化技术检测150对食管鳞癌组织中HMGA2 蛋白的表达，分析let-7表达水平与食管鳞癌临床病理参数的关系。 结果：转染let-7 的Eca109细胞中，let-7表达水平升高约33%，并且抑制细胞的增殖，同时HMGA2蛋白的 表达水平下降约30％；转染let-7抑制剂的Eca109细胞中，let-7表达水平下降到50%， 并且促进细胞的增殖，HMGA2蛋白的表达水平增加约64％；然而，各组中HMGA2 mRNA 水平并未受影响。Let-7在食管鳞癌组织中的表达显著低于对应的癌旁正常食管鳞状 上皮组织（P＜0.05），其表达水平与淋巴结转移相关；HMGA2 mRNA水平虽然在癌组织 与癌旁组织间无明显差异（P＞0.05），但其蛋白水平在癌组织中的表达显著高于对应 的癌旁正常食管鳞状上皮组织（P＜0.05）。 结论：1. Let-7、HMGA2参与了食管鳞癌 的发生；2. Let-7能够抑制食管鳞癌细胞的增殖，let-7与HMGA2蛋白的表达呈反相关； 3. Let-7在食管鳞癌组织中呈低表达，且其低表达与淋巴结转移相关；而HMGA2在癌 组织中呈高表达，let-7可能在转录后水平调控HMGA2蛋白的表达。 关键词：食管鳞癌；微小 RNA let-7；HMGA2 Role of microRNA let-7 and effect to HMGA2 in esophageal squamous cell carcinoma Postgraduate:Qing Liu Supervisor: Professor Xiaomei Lu Abstract Objective To investigate the role of microRNA (miRNA) let-7 and its regulation on high mobility group A2 (HMGA2) protein expression in esophageal squamous cell carcinoma (ESCC). Methods Chemical synthesis of let-7 or its inhibitor were transfected into a human ESCC cell line Eca109. The expression of let-7 and HMGA2 mRNA in the transfected cells were detected by qRT-PCR method, and the protein expression of HMGA2 were examined by western blot technique. Specimens of ESCC and normal tissues were collected from ESCC patients, the expression of let-7 and HMGA2 mRNA were detected by qRT-PCR method, and the protein expression of HMGA2 was examined by immunohistochemistry (IHC). Results In the Eca109 cells transfected with let-7, the increased accumulation of let-7 repressed cells proliferation, and resulted in decrease of HMGA2 protein levels with approximately 30％. However, in cells transfec