口腔鳞癌细胞中mirna-mirna in oral squamous carcinoma cells.docxVIP

目录中文摘要··············································································1英文摘要··············································································4英文缩写··············································································7研究论文口腔鳞癌细胞中miRNA-222与CDKN1B(p27kip1)mRNA及靶蛋白表达关系的研究前言··············································································8材料与方法·····································································11结果··············································································20附图··············································································21附表··············································································24讨论··············································································25结论··············································································29参考文献········································································30综述MicroRNA与口腔鳞状细胞癌的研究进展····························34致谢····················································································47个人简历··············································································48口腔鳞癌细胞中miRNA-222与CDKN1B(p27kip1)mRNA及靶蛋白表达关系的研究摘要目的：口腔鳞癌（oralsquamouscellcarcinomas，OSCC）是口腔颌面部常见的恶性肿瘤之一，约占颌面部恶性肿瘤的80%以上，在全身范围的恶性肿瘤中居第八位[1]。近年来，口腔鳞癌的发病率呈逐年上升的趋势。目前临床上对口腔鳞癌的治疗方法以手术扩大切除为主，辅以放疗、化疗、生物治疗、靶向治疗、免疫治疗等。虽然对于口腔鳞癌的治疗手段实现多元化及精准化，但因其易发生局部复发及淋巴转移和远端转移而影响口腔鳞癌患者的预后。因此深入了解口腔鳞癌的发病机制、早期诊断及预后分析显得十分迫切。研究证实多种原癌基因及抑癌基因的异常表达可能与口腔鳞癌的发生发展密切相关。miRNA是一类内源性非编码小RNA，长约18～23nt。miRNA因其非编码特性可在翻译水平或者转录后水平调控一个或多个靶基因mRNA及靶蛋白的表达水平［2］。CDKN1B（p27kip1）是近来新发现的一种肿瘤抑制基因，其是细胞周期素依赖性激酶抑制蛋白CDKI家族的最新成员，其与细胞周期关系密切，可以阻断细胞从G1期到S期。本研究通过对人舌鳞癌细胞株TCA-83瞬时转染miRNA-222模拟物及抑制物，同时设立阴性对照，分析其靶基因CDKN1B（p27kip1）mRNA差异性表达及其靶蛋白在口腔鳞癌细胞中的表达差异，探讨miRNA-222对口腔鳞癌细胞中CDKN1B（p27kip1）的靶向调控作用，以期对口腔鳞癌的早期发现、准确诊断、靶向治疗等提供一定的理论基础。方法：1通过LipofectamineTM2000转染试剂将人工合成的miRNA-222mimics（miRNA-222上调组）和inhibitor（miRNA-222下调组）转染人舌鳞癌细胞株TCA-83，同时设立阴性对照组及空白对照组。应用反转录-聚合酶链式扩增技术（RT-PCR）分别检测miRNA-22