【作者】 马全；

【导师】 武正炎；

【作者基本信息】 南京医科大学 ， 普外科， 2005， 博士

【摘要】 背景 自1991年发现第一个黑色素瘤相关抗原基因(Melanoma-associated Antigen Gene,Mage)及其抗原肽(MZ2-E)以来,多年来,人们已在不同肿瘤组织中,发现多个Mage基因,均定位于X染色体,有着一定的同源性,组成了一个Mage基因家族,分为A、B、C、D、E、F六个亚家族。进一步的研究发现,其中A、B、C三个亚家族存在一些独特的特征,只在肿瘤、睾丸和胎盘组织中表达,在其它正常组织迄今为止还没有发现其表达。而且其表达产物是一种肿瘤排斥性抗原,细胞毒性T淋巴细胞(cytotoxic T lymphocyte,CTL)可以识别该抗原。这提示Mage基因有可能作为肿瘤相关的检测标志,有助于肿瘤的诊断与治疗,而其抗原有可能做为肿瘤免疫治疗的靶抗原进行免疫治疗。 树突状细胞(dendritic cells,DCs)是一类形态独特的MHC携带细胞,有强有力的免疫刺激力和抗原递呈能力,也是唯一能激活初始型T细胞、诱发初次免疫应答的抗原递呈细胞。越来越多的证据表明,DC诱发的细胞免疫尤其是CTL介导的免疫应答,在消除恶性肿瘤中起着重要作用。利用树突状细胞负载抗原进行细胞过继免疫治疗(adoptive cellular immunotherapy,ACI)受到广泛关注。 基于Mage基因及其抗原肽的特征,我们对Mage基因在乳腺癌中的表达,在隐匿肿瘤细胞(occult tumor cells,OTC)检测中的价值,以更多还原

【Abstract】 Background:Since van der Bruggen found the first melanoma-associated antigen gene (Mage) and its peptide MZ2-E in 1991, the researchers had found dozens of Mage in many different types of tumors. They were all mapped to chromosome X and shared certain homologous regions. They were all the members of Mage family, including six subgroups A、 B、 C、 D、 E、 F. And the subgroups A、 B. C shared certain homology. They has been found in many different types of malignant tumors, but not in normal adult tissues except the testis and placenta. Though scientists didin’t know the physiological function of Mage genes encoding proteins, researchers had found that they were tumor antigens which could be recognized by specific T cells in malignant tumor. It suggeated us Mage maybe a novel marker to detect tumor and its peptide may be the most promising candidate for tumor-specific immunotherapy of cancer.Dendritic cells (DCs) were the most potent and the only antigen presenting cells (APC), which were capable of activating naive T cells and initiating primary immune response. Recently, more and more evidence showed that the cellular immunity induced by dendritic cells, especial theCTL response, played an important role in the immune response against the malignant tumor. The adoptive cellular immunotherapy based on the dendritic cells loaded with antigen is becoming the highlight in the biology therpy of malignant tumor.The expression of Mage-A1 、 A2、 A3 in breast cancer tissues and four breast cancer cell lines was researched. Their mRNA was used as the marker to detecte the occult tumor cells in peripheral blood of patients with breast cancer. The experimental immunotherapy based on dendritic cells loaded with Mage-A3 peptide were studied on the mice model of breast cancer. 1. Expression of Mage-A1、 A2、 A3 in breast cancer tissue and breast cancer cell linesObjectives To study the expression of Mage-A1、 A2、 A3 in breast cancer tissues and four breast cancer cell lines.Methods The expression of Mage-A1、 A2、 A3 in breast cancer tissues and four breast cancer cell lines, MCF-7、 Sk-Br-3、 MDA-MB-435s and TM40D was detected by reverse transcription polymerase chain reaction (RT-PCR) . Results The positive expression rate of Mage-A in breast cancer tissues was 13/33 (39%) , of Mage-Al was 4/33 (12%) , of Mage-A2 was 8/33 ( 24%), and of Mage-A3 was 7/33( 21%), respectively. Both Mage-A1 and Mage-A3 were positive in breast cancer cell lines MCF-7 and Sk-Br-3. MDA-MB-435s expressed Mage-A2 and Mage-A3. Mage-A3 was positive in TM40D.Conclusions Mage genes were often expressed in breast cancer, butexpression of Mages varied in the breast cancer cell lines. Mage geneencoding proteins are eligible for Mage-peptide-based activeimmunotherapy.2. Expression of Mage-A1、 A2、 A3 in the occult tumor cells in theperipheral blood of patients with breast cancerObjective To detect tumor cells in the peripheral blood of patients withbreast cancer by using the mRNA of the Mage-A1 and Mage-A3 genes asspecific tumor markers.Methods Peripheral blood was obtained from 40 breast cancer patients and20 patients with benign diseases. The mRNA of the Mage-A1 and Mage-A3genes in the peripheral blood mononuclear cells (PBMCs) was detected bynested RT-PCR. The Mage-A1 and Mage-A3 transcripts in the tumor tissuesof these breast cancer patients were also detected by RT-PCR.Results Of the 40 breast cancer patients, Mage-Al and Mage-A3 mRNAwere positive in 12.5% ( 5/40) and 17.5% ( 7/40 ) of PBMCs respectively, andin 15 % ( 6/40 ) and 22.5 % ( 9/ 40 ) of breast cancer tissues respectively. Inthe PBMCs of the 40 breast cancer patients, 10( 25 % )samples were detectedto express at least one type of Mage mRNA. Mage mRNA were not found inthe PBMCs from the patients whose tumors did not express the Mage genes,nor in the PBMCs from the 20 patients with benign diseases. The positiverate of Mage mRNA in the PBMCs was closely correlated with the TNMstages.Conclusion Mage-A1 and Mage-A3 mRNA could be specificallydetected in the PBMCs of breast cancer pati更多还原