寡核苷酸基因芯片检测肺癌中点突变的研究;二氧化硒诱导肺癌细胞凋亡过程中c-fos蛋白表达的改变-内科呼吸专业论文.docxVIP

中毽耱器l鲑辩大学薅圭争瓶逶交第一部分寡核苷酸基因芯片检测肺癌中点突变的研究 中毽耱器l鲑辩大学薅圭争瓶逶交 第一部分寡核苷酸基因芯片检测肺癌中点突变的研究 第一章Rb2／p130寡核营酸基因蕊背靛制备及赫癌手术标本酶检测 中文攘要 目的：用寡核蛩蔽基因芯片的方法，在中豳繁次检测肺癌标本中Rb2／p130綦因的热点 突变，为S乖癌韵诊断撵索快速、准确鹪基因捡测途径。 方法：1．枝巢耱瘿患者麓手术标本，并援取藻嚣疆DNA。 2．在Gene bank中检索Rb2／p130基因的DNA序列后，设计并合成探针和引物序 裂。 3．蒋搽锋按一定瓣蔟浮点弹子醛蕊纯瓣玻冀上，臻l蛰或蘩因芯冀。 4，霜荧竞撩记鹩引物对黼癌标本DNA遗芎亍Exon 21、Exon 22的PCR扩增，然 后用PCR扩增产物与基因芯片进行杂交。 5．逶过黠杂交髂号懿羧测，蘩辩楚否发囊燕点突交，势蕊灞黪鹃方法对突变甓意 进行骏避。 结果：本实验月寡核萤酸綦因芯片豹方法检测了30侧月帝癌标本的Rb2／p130基躐靛突 变。缝累裳霹，PCR产耪浓囊秘杂交滋度是澎嫡结巢的2个关键医豢。在3移镶 肺癌标本中，Rb2／p130基因突变的梭出率是16。67％，其中，肺腺癌的突褒率是 20％(4／20)，§漆鳞癌的突交率16、67％(t翰，Exon 2t突嶷4铡次，Exon 22突变t 蜘次。对突变往点的颡l序证实了鏊麓芯片静缭巢。 结论：寡核脊黢基因芯片法检测出中国人肺癌组织桥本存在Rb2／p130基因突变，但突 变事≤16+爵7翰甄予谣方人(78，s％)。未鼹糗溺奎帮突变馥熹、猿零塞枣器尺黪麓舅 可能趋低突变率的原鳓。 美键词：肺瘸，基因芯片，Rb2／p130基因，点突变 中霉协和医辩大学博士学位论文Abstract 中霉协和医辩大学博士学位论文 Abstract objective：Part of the hot-spot mutations of Rb2／p130 gene was screened in resected lung cancers using oligonucleotide gene chips in order to detect rapid and accurate method in gene diagnosis of lung cancer． Methods：Samples of resected lung cancers were collected and genomic DNAs were extracted．After DNA sequence of RHb2／p130 was analysed，probes and primer were synthesized．Gene chips were made by doting the probes to modified slide。Using primer marked by fluorescence，Exon 21 and Exon 22 PCR amplification of Rb2／D130 were performed，then hybridization of the products of PCR amplification and gene chips were used．Hybridization signal was detected in o砖er to assess hot-spot mutation，Finally sequencing was performed to confirm the hot-spot mutation． Results：Using oligonucleotide gene chips，we screened the Rb2／p130 gene mutation in 38 sample of kng cancer．We conclude that the concentration of PCR production and hybridization temperature are the key factors affecting the results．Of the 30 patients，16．67％with Rb2／p130 matations．4(4／20)mutations happened in adenocarcinoma，1(I／6)mutations happened in squamous carcinoma。 Sequencing map confirmed the mutation ofl№2，p130 in lung cancer sample． Conclusion：Rb2／p130