肝癌组织中三个MAGE家族基因的克隆

目的:从肝癌组织中克隆肿瘤相关抗原基因MAGE-1的全长cDNA,为该基因及其编码抗原应用于肝癌的免疫治疗奠定基础.方法:根据MAGE-1编码区上、下游序列设计一对引物,用RT-PCR方法从肝癌组织中扩增该基因的全长cDNA,酶切产生粘性末端后连接入PUC19质粒.经初步酶切鉴定后,通过DNA序列分析获取所克隆基因片断的核苷酸序列.结果:成功地克隆了MAGE-1基因的全长cDNA,同时还获得一约750bp的MAGE-3基因片段及一与MAGE-6和MAGE-12高度同源的基因的克隆.此与MAGE-6,-12高度同源的基因可能为一未知MAGE家族成员.结论:肝癌组织中表达多种MAGE家族基因,甚至可能存在未知MAGE家族基因的表达,这将为寻找肝癌免疫治疗的攻击靶点开辟新的途径.     

MAGE的研究现状及展望

恶性肿瘤是当代威胁人类健康的最严重的疾病,具有病期越晚治疗越难、预后越差的特点,因此寻找肿瘤特异性抗原(tumor-specific antigen,TSA)和肿瘤相关性抗原(tumor-associated antigen,TAA)对于肿瘤的早期发现、早期诊断和早期治疗极为重要。     

Cystatin家族的研究进展

近几年来，随着对肿瘤发病机理研究的不断深入，发现Cystatin家族作为半胱氨酸蛋白酶的内源性抑制剂，与肿瘤的发生、发展、浸润和转移关系非常密切，有可能成为临床诊断和预后检测的分子指标。 1 Cystatin家族的发现 　　1983年AnastasiA等[1]采用亲和层析的方法首次在鸡蛋清中分离得到了两种不同pI值(6.5和5.6)的蛋白质，接下来的研究发现，它们对半胱氨酸蛋白酶(cysteineprotease)有抑制作用，因而把它命名为Cystatin；1984年DaviesME等把CystatinA、CystatinB分别定位在人体肝脏中性粒细胞和淋巴细胞中；同年BarrettAJ等在脑垂体瘤病人肿瘤细胞的细胞质中发现CystatinC。自此以后，Cystatin家族成员陆陆续续在不同的物种中被分离得到，现在已经形成了一个非常庞大的超家族。     

Caspase家族与细胞凋亡的研究进展

近年来Caspase蛋白酶家族是细胞凋亡研究的热点,Caspase被认为是细胞凋亡的中心环节和执行者.现就Caspase蛋白酶家族生物学特性、分布、作用底物、抑制剂,以及激活机制、凋亡调控基因之间的作用等从分子机制对凋亡调控的研究现状及与肿瘤和疾病防治可能的应用前景进行综述.     

MAGE3 siRNA表达载体的构建及其沉寂作用

目的构建针对人MAGE3基因的小干扰RNA(siRNA)及其表达载体,转染细胞后观察其对MAGE3基因的干扰作用。方法设计MAGE3靶向的发夹状siRNA,依据设计合成两条互补的寡核苷酸链,退火后连接入pSUPER.neo载体,转化扩增后进行序列测定。用脂质体包裹转染胃癌细胞BGC823,采用RT-PCR检测MAGE3基因mRNA表达水平的变化。结果把针对MAGE3基因的siRNA的双链寡核苷酸片段克隆到pSUPER.neo载体,经过酶切鉴定与测序,结果正确;稳定转染人胃癌细胞BGC823后,RT-PCR检测显示,MAGE3基因的表达水平明显降低。结论成功构建了针对MAGE3基因的siRNA载体,转染细胞后可抑制MAGE3基因表达。     

MAGE3-HLA-A2肽疫苗诱导胃癌患者自体DC及激活CTL杀伤性的研究

本实验研究MAGE3-HLA-A2肽疫苗诱导胃癌特异性CTL的杀伤作用,及免疫治疗的可行性.选择HLA血清学分型试剂盒购自(中日友好医院),RT-PCR试剂盒购自日本TAKARA公司,MAGE3引物序列由日本TAKARA公司合成,41例胃癌患者来自本院腹部外科,血细胞分离仪采用CS3000PLUS.MTT试剂盒(德国宝灵曼公司),靶细胞系Mel-526(表达MAGE3-HLA-A2)由Mr.Dr TaKesako惠赠,用胃癌患者外周血5ml采用血清学方法检测HLA-A分型,筛选HLA-A2阳性患者留取新鲜胃癌手术标本1cm~3,RT-PCR方法检测MACF3表达,MACE3/HLA-A2阳性患者经CS-3000PLUS采集单个核细胞,GM-CSF IL4诱导DC7d,流式细胞仪检测     

MAGE3 siRNA表达载体的构建及其沉寂作用

目的构建针对人MAGE3基因的小干扰RNA（siRNA）及其表达载体，转染细胞后观察其对MAGE3基因的干扰作用。方法设计MAGE3靶向的发夹状siRNA，依据设计合成两条互补的寡核苷酸链，退火后连接入pSUPER．neo载体，转化扩增后进行序列测定。用脂质体包裹转染胃癌细胞BGC823，采用RT-PCR检测MAGE3基因mRNA表达水平的变化。结果把针对MAGE3基因的siRNA的双链寡核苷酸片段克隆到pSUPER．neo载体，经过酶切鉴定与测序，结果正确；稳定转染人胃癌细胞BGC823后，RT-PCR检测显示，MAGE3基因的表达水平明显降低。结论成功构建了针对MAGE3基因的siRNA载体，转染细胞后可抑制MAGE3基因表达。     

Caspase家族与细胞凋亡的研究进展

近年来Caspase蛋白酶家族是细胞凋亡研究的热点，Caspase被认为是细胞凋亡的中心环节和执行者。现就Caspase蛋白酶家族生物学特性、分布、作用底物、抑制剂，以及激活机制、凋亡调控基因之间的作用等从分子机制对凋亡调控的研究现状及与肿瘤和疾病防治可能的应用前景进行综述。     

姜黄素治疗甲状腺癌的相关机制研究进展

近年来中国的甲状腺癌(TC)发病率呈上升趋势.姜黄素是从植物姜黄中提取出来的一种天然的多酚,可以抑制多种肿瘤细胞的生长,具有应用活性良好且安全无毒的天然活性成分,是目前治疗肿瘤研究的热点.近来的体外和体内研究相继证实姜黄素可以诱导恶性肿瘤细胞分化及肿瘤细胞凋亡、抑制肿瘤细胞周期、抑制肿瘤细胞的黏附与浸润、血管的生成和淋...     

宫颈癌相关细胞凋亡通路研究进展

目前宫颈癌治疗仍以手术切除、放疗和化疗等传统手段为主。晚期宫颈癌易复发、难以根治。细胞凋亡信号通路受阻是包括宫颈癌在内的恶性肿瘤最大特征之一。也是宫颈癌产生抗药性、复发和转移而难以治愈的主要原因。恢复癌变细胞凋亡信号通路,诱导癌细胞走向凋亡将从根本上解决宫颈癌难以控制的困扰。本文对近10年来关键细胞凋亡信号通路在宫颈癌中的应用研究进展进行综述,进一步明确通过诱导细胞凋亡在宫颈癌治疗中的作用。     

Effects of a novel antitumor depsipeptide, FR901228, on human breast cancer cells

Human breast cancer MCF7 and MDA-MB231 cells were used to investigate the biological and molecular activities of a novel naturally occurring agent, FR901228 (FR), that possesses a potent antitumor activity against human and murine tumor cells. Investigation of the cytotoxicity of FR and induction of internucleosomal DNA degradation in FR-treated cultures revealed that FR induced apoptotic-like cell death of MCF7 and MDA-MB231 cells. In FR-treated apoptotic cultures, flow cytometry revealed that there was a significant decrease of cells in S phase of the cell cycle. In FR-treated cells there was an increased expression of p21Cip1 and phosphorylation of Bcl-2 as determined by Western immunoblotting, and a novel cytoplasmic kinase of 33 kDa, p33 kinase, as determined by the in-gel kinase assay using myelin basic protein (MBP) as a substrate. Increased expression of p21Cip1, phosphorylation of Bcl-2, and activation of p33 MBP kinase may play part of the key mechanism for FR-induced apoptosis.     

Pathogenicity of the MAGE family

The melanoma antigen gene (MAGE) protein family is a group of highly conserved proteins that share a common homology domain. Under normal circumstances, numerous MAGE proteins are only expressed in reproduction-related tissues; however, abnormal expression levels are observed in a variety of tumor tissues. The MAGE family consists of type I and II proteins, several of which are cancer-testis antigens that are highly expressed in cancer and serve a critical role in tumorigenesis. Therefore, this review will use the relationship between MAGEs and tumors as a starting point, focusing on the latest developments regarding the function of MAGEs as oncogenes, and preliminarily reveal their possible mechanisms.     

Gankyrin在炎症相关性癌症中的作用机制研究进展

慢性非可控性炎症是癌症发生发展的基本微环境。约1/4癌症病例的病因和发病机制与感染和慢性炎症相关。Gankyrin是一个重要的癌蛋白,它在多种恶性肿瘤中高表达,并在癌症发生发展中起着关键的作用。近期研究发现Gankyrin在结肠炎相关性结肠癌（colitis-associated cancer,CAC）和慢性肝炎向肝细胞肝癌（hepatocellular carcinoma,HCC）发展的过程中扮演着重要的角色。同时,Gankyrin与多种细胞因子以及炎症信号通路密切相关。本文对Gankyrin在炎症相关性癌症中的作用机制研究进展作一简要综述。     

Expression of the MAGE gene family in human hepatocellular carcinoma

BACKGROUND: The 12 members of the MAGE gene family encode tumor specific antigens that are recognized by autologous cytotoxic T lymphocytes (CTL). The MAGE genes are expressed not only in melanoma but in the other malignant tumors as well. There is, however, little information on their expression in hepatocellular carcinoma (HCC). The authors thus studied the expression of the MAGE gene family in human HCC and discuss the possibility of specific immunotherapy using MAGE peptides. METHODS: Tumor tissue samples of HCC and paired nontumor tissues of the liver were obtained from 22 HCC patients. Total RNA was extracted and cDNA was synthesized. Polymerase chain reaction amplification using each MAGE gene specific primer was then performed to detect the expression of each MAGE gene. Immunoblotting and immunohistochemical analysis were performed to confirm the expression of MAGE-3 gene product in HCC. RESULTS: The expression rate of each MAGE gene was as follows: MAGE-1 and -3 were expressed in approximately 68% of the tumors; MAGE-8 was expressed in 46%; and MAGE-2, -6, -10, -11, and -12 were expressed in approximately 30%. Nineteen (86%) of 22 tumors expressed at least 1 MAGE gene. On the other hand, no expression was detected in the noncarcinomatous liver tissue specimens. Actual expression of the gene product of MAGE-3 was detected in 50% of tumors. Clinicopathologic data on the MAGE positive and negative cases were compared. Significant differences were observed between MAGE expression status and a few clinicopathologic factors; however, further investigation is required to elucidate these correlations completely. CONCLUSIONS: These findings demonstrated that MAGE gene expression is frequent in HCC, thus suggesting that HCC patients may be good candidates for specific immunotherapy using MAGE peptides.     

胰腺癌癌前病变基因研究进展

现已证实,基因研究有助于阐明能够促进胰腺癌（pancreatic cancer, PC）发生、发展的一些关键基因和分子通路。早期研究发现,在胰腺上皮内瘤样变（PanINs）进展过程中,随着组织异型程度的增加,相应的发生KRAS基因激活,CDKN2A基因、TP53基因及SMAD4基因失活,并可导致多个重要的细胞过程和信号转导通路发生改变。而在导管内乳头状黏液性瘤（IPMNs）和黏液性囊性瘤（MCNs）进展过程中,也发生了相应的基因突变。最近,全基因组外显子测序技术揭示了由癌前病变进展而来的胰腺癌基因图谱,这些研究对发展形成新的个体化治疗方法有所帮助。本文从胰腺三种癌前病变形态学变化、各自的基因突变事件以及胰腺癌前病变中部分信号通路和调控过程与基因突变的关系等方面作一综述。     

Differential expression profile of MAGE family in non-small-cell lung cancer

The expression of the melanoma-associated antigen (MAGE) genes consists of variables in all tumor types, such as lung cancer, which are relevant to be silent in all normal tissues except germ cells. They are considered as tumor-specific antigens, and are ideal targets for cancer immunotherapy. A complete MAGE genes differential expression profile analysis of lung cancer can provide this study not only various target genes for immunotherapy, but also valuable markers for further diagnosis and prognosis. This research has constructed a membrane array, which was consisted 32 MAGE genes, to detect whether the differential expression profile occurred in 52 pairs of non-small-cell lung cancer (NSCLC) samples. Nearly 32 MAGE genes have been differential expressed in NSCLC except MAGE-B1 and -E2. MAGE-B, -C, -D, and subgroup -B6, -D4 have showed prominences in lung adenocarcinoma. High-frequent expression of MAGE-D, and subgroup -A2, -D2 has also been discovered in non-metastasis group (p<0.05). However, there is no significant difference of MAGE genes differential expression shown among different primary tumor (T), nodal involvement (N) and overall stages. Several MAGE subgroup genes, such as MAGE-A5, -A7, -A8, -A9, -A11, -B3, -B4, -B10, -D2, -D3, -F1, -G1, -H1, and -L2, have been first discovered to show differential expression in NSCLC. Although the small size of the sample may limit the diagnostic and prognostic value of MAGE genes, the function of the membrane array can provide this study a high-throughput method to detect the whole MAGE genes differential expression profile.