【作者】 郭明明；

【导师】 余之刚；

【作者基本信息】 山东大学 ， 外科学， 2011， 硕士

【摘要】 研究背景目前乳腺癌已经在世界范围内成为严重威胁女性健康的重大恶性疾病之一。上世纪80年代以来,在全球范围内乳腺癌分发病率都呈现出非常明显的上升趋势,每年以2%-3%的速度递增,而这一现象在发展中国家尤为突出。有数据显示,2000-2005年,中国乳腺癌发病人数增长38.5%,死亡人数增长37.1%。目前乳腺癌发生发展过程尚不完全明了,从而在一定程度上对乳腺癌的早期诊断、相关治疗带来很大的困惑。任何肿瘤的发生发展都包含了十分复杂的生物学过程。对个体而言,所有机体细胞携带相同的遗传模板,那么各种细胞型,包括癌细胞在内,是如何获得不同的表型的呢?科学家最终发现,各种类型细胞对基因组的选择性表达决定了细胞的不同表型,同时提示我们基因表达的异常改变是正常细胞向恶性肿瘤细胞转变的基础。任何肿瘤都起源于正常组织,肿瘤细胞是由失去了聚集能力和构成正常组织能力的细胞构成的。RNA,是连接DNA与蛋白的桥梁。通过分析RNA水平的差异,能够为我们下一步在分子水平研究相关基因／蛋白在疾病发生发展过程中的作用提供理论依据,也是寻找疾病相关基因／蛋白的一种理想方法。因此通过基因表达谱芯片比较乳腺癌及正常乳腺组织的差异表达基因,并据此进行相关生物学过称与乳腺癌发生发展之间的关系,将对明确乳腺癌的生物学过程提供一定的研究基础。研究目的本次研究旨在探索乳腺癌组织与自身正常乳腺组织、乳腺良性疾病之间的差异表达基因,为今后进一步研究相关基因与乳腺癌的关系,相关基因或其表达产物在乳腺癌发生发展过程中的作用打下基础。研究方法本研究采用全人类基因组芯片,检测7对乳腺癌及其正常乳腺组织、7例乳腺良性疾病的差异表达基因,通过real-time PCR对芯片试验结果进行验证,并根据GeneOntology(GO)分类体系初步对得出的差异表达基因进行功能分类。结果1.差异表达基因：乳腺癌组织同癌旁正常乳腺组织相比,共得出344个差异表达基因。在这一组基因中,上表达的有273个,下表达的有71个(与细胞增殖有关的基因有27个,其中上表达的有21个,下表达的有6个；与细胞粘附有关的基因有15个,其中上表达的有5个,下表达的有10个；与细胞凋亡有关的基因有23个,其中上表达的有20个,下表达的有3个)。乳腺癌组织与乳腺良性疾病的差异表达基因有235个,上表达的有140个,下表达的有95个(与细胞增殖有关的基因有10个,其中上表达的有8个,下表达的有2个；与细胞粘附有关的基因有12个,其中上表达的有7个,下表达的有5个；与细胞凋亡有关的基因有11个,其中上表达的有10个,下表达的有1个)。2.乳腺癌和正常乳腺组织的聚类分析：采用cluster 3.0软件对所得实验数据进行聚类分析,可以发现三种乳腺组织不能完全分开。单纯分析7对乳腺癌和癌旁正常乳腺组织,聚类分析图显示二者也不能够完整的分开。结论乳腺癌组织与正常乳腺组织间在基因的表达层面存在明显差异。筛选得到的基因涉及细胞增殖、细胞粘附、细胞凋亡等肿瘤发生发展的重要生物学过程,可能与乳腺癌的发生发展密切相关,但仍需要针对兴趣基因进行更深入的分子水平的研究。对于浩如烟海的基因信息而言,为感兴趣的生物学过程设定这一个限制,在有限的范围内比较相关基因的功能、细胞定位及其表达水平的高低,将有利于发现其中功能上有相互联系的基因,为选择出兴趣基因进行基因或蛋白水平的研究提供一定的线索。更多还原

【Abstract】 BackgroundBreast cancer has became one of the major malignant diseases which threaten women’health seriously in the worldwild. Since 1980s, the incidence of breast cancer has showed an upward trend with annual increasing rate of 2%～3% in the world. This phenomenon is worse especially in the developing countries than the developed countries. Data has shown that the number of new cases of breast cancer in China has increased by 38.5% while the death number of breast cancer has increased by 37.1% from 2000 to 2005. Currently, the oncogenesis of breast cancer is not very clear, so there are still a lot of confusion in the early detection and treatment of breast cancer. Any oncogenesis is a very complicated biological process. For an individual, the cells of almost the whole body have the same genetic material DNA, why they (including the tumor cells) have different phenotypes? Data show that the selective expression of genome decides the phenotype of different cells. This indicates that the abnormal change of genetic expression is the foundation by which process the normal cell change into tumor cell. All tumors originate from normal tissues. Tumor cells are the cells which lose the ability of aggregation and constituting the normal tissues. RNA is the connection of DNA and protein. By analyzing the difference of RNA levels between different tissues, we can find some theoretic clue for the further study of gene/protein involved in the ontogenesis of some diseases. This is also a good way to find some related genes or protein in some diseases. So comparing breast cancer and normal breast tissue by microarray to find diffenently expressed genes and to do further study will do some help in clearing and definiting the oncogenesis of breast cancer. This study is aimed to explore the differently expressed genes between breast cancer and normal breast tissue, can provide the evidence of the related genes/proteins to breast cancer’development.ObjectiveExplore the differently expressed genes between breast cancer and normal breast tissue, explore the differently expressed genes between breast cancer and benign breast disease tissue, to provide some guideline for the study of oncogenesis of breast cancer development.MethodsIn this study,21 breast tissues (7 are breast cancer tissues,7 are their corresponding normal breast tissues,7 are benign breast disease tissues) were detected by microarray. Then we used real-time PCR to verify the results of microarray. We used GeneOntology (GO) system to classify the differently expressed genes.Results1. Differently expressed genesThere are 344 differently expressed genes between breast cancer tissue and corresponding normal breast tissues, including 273up-regulated and 71 down-regulated genes. There are 27 genes related to cell proliferation, while 21 up-regulated and 6 down-regulated. Fifteen genes are related to cell adhesion, while 5 up-regulated and 10 down-regulated. Twnety three genes are related to cell apoptosis, while 20 up-regulated and 3 down-regulated.There are 235 differently expressed genes between breast cancer tissue and corresponding normal breast tissues, including 140 up-regulated and 95 down-regulated genes. There are 10 genes related to cell proliferation, while 8 up-regulated and 2 down-regulated. Twelve genes are related to cell adhesion, while 7 up-regulated and 5 down-regulated. Elevne genes are related to cell apoptosis, while 10 up-regulated and 1 down-regulated.2. Cluster analysis of the two kinds of samplesUsing cluster 3.0 to analyze the data of microarray experiment, the 3 kinds of samples can not be divided to 3 groups directly. To analyze the data of microarray experiment of breast cancer and correspongding normal breast tissue, the result is the same.ConclusionThere are obviously difference in the level of mRNA between breast cancer and normal breast tissue. The differented expressed genes in this study refered to some important biological process in the development of cancer, such as cell proliferation, cell adhesion and cell apoptosis. These genes may have some relationship with breast cancer. We still need more study to verify the function of these genes in ontogenesis of breast cancer. By this study, we can limit the related genes for breast cancer from mass of gene information. We can study the related function between these genes, find some candidate genes to study their change in gene or protein in the development of breast cancer.更多还原