癌-睾丸抗原MAGE-A1、A3、A4在视网膜母细胞瘤中的表达

目的了解视网膜母细胞瘤（RB）组织癌—睾丸抗原（CTA）基因MAGE-A1、A3、A4的表达,探讨这些基因成为RB特异性免疫治疗靶抗原的可能性。方法人RB组织15例,对照组为12例非肿瘤病变视网膜组织及22例正常眼部组织。采用逆转录－聚合酶链反应（RT-PCR）技术检测上述组织MAGE-A1、A3、A4的mRNA表达情况,并根据肿瘤病理分级、肿瘤大小、临床分期等临床指标进行统计学分析。结果15例RB组织中,MAGE-A1、A3、A4的表达率分别为33.33%（5/15）、46.67%（7/15）和33.33% （5/15）,有73.33%（11/15）的RB组织至少表达一种MAGE基因,33.33%（5/15）的RB组织至少表达两种MAGE基因。12例非肿瘤病变视网膜组织及22例正常眼部组织三种MAGE基因均不表达。结论MAGE-A1、A3、A4基因在RB组织中有一定频率的表达,MAGE-A1、A3、A4的表达率与病人的性别、年龄和临床分期,肿瘤大小及病理分级均无相关性（P＞0.05）。MAGE-A1、A3、A4有希望成为RB特异性免疫治疗的靶抗原。     

肿瘤-睾丸抗原SSX1和SSX4的mRNA在肝细胞肝癌中的表达

目的检测肿瘤-睾丸抗原(Cancer-Testis antigens,CT)SSX1和 SSX4基因 mRNA 在肝细胞肝癌(HCC)中的表达,探讨 CT 抗原 SSX1和 SSX4 mRNA 在 HCC 中表达有无特异性。方法应用逆转录-聚合酶链反应(RT-PCR)方法对35例 HCC 患者癌组织和相应的癌旁组织 SSX1和 SSX4基因表达进行检测,对其中6例 RT-PCR 扩增产物的目的片段进行 DNA 序列测定。结果 35例 HCC 患者中,SSX1和 SSX4的表达率分别为81%(27/35)和73%(23/35);癌旁组织、12例肝硬化和15例正常组织未检测到 SSX1和 SSX4的表达。6例 DNA 序列测定结果显示 RT-PCR 产物为 SSX1和 SSX4基因的 cDNA。SSX1和 SSX4的表达与患者年龄、性别、肿瘤大小、分化程度、血清甲胎蛋白(AFP)水平、乙型肝炎病毒(HBV)和丙型肝炎病毒(HCV)感染无显著相关性(P>0.05)。结论 SSX1和 SSX4在 HCC 中呈高特异性表达,是制备 HCC 疫苗理想的靶向分子,多种 CT 抗原联合表达为多效价疫苗治疗 HCC提供了理论基础。     

肿瘤-睾丸抗原SSX1和SSX4的mRNA在肝细胞肝癌中的表达

目的 检测肿瘤-睾丸抗原(Cancer-Testis antigens，CT)SSX1和SSX4基因mRNA在肝细胞肝癌(HCC)中的表达，探讨CT抗原SSX1 和SSX4 mRNA在HCC中表达有无特异性。 方法 应用逆转录-聚合酶链反应(RT-PCR)方法对35例HCC患者癌组织和相应的癌旁组织SSX1和SSX4基因表达进行检测，对 其中6例RT-PCR扩增产物的目的片段进行DNA序列测定。 结果 35例HCC患者中，SSX1和SSX4的表达率分别为81％(27／35)和73％(23／35)；癌旁组织、12例肝硬化和15例正常组织未 检测到SSX1和SSX4的表达。6例DNA序列测定结果显示RT-PCR产物为SSX1和SSX4基因的cDNA。SSX1和SSX4的表达与患 者年龄、性别、肿瘤大小、分化程度、血清甲胎蛋白(AFP)水平、乙型肝炎病毒(HBV)和丙型肝炎病毒(HCV)感染无显著相关性(P >0．05)。 结论 SSX1和SSX4在HCC中呈高特异性表达，是制备HCC疫苗理想的靶向分子，多种CT抗原联合表达为多效价疫苗治疗HCC 提供了理论基础。     

肿瘤睾丸抗原(CTA)在肺癌中的表达

目的：检测多种肿瘤睾丸抗原（CTA）基因在肺癌中的表达。方法：用逆转录－聚合酶链反应（RT－PCR）法对35例肺癌患者癌组织和癌旁正常肺组织进行MAGE－1、－3，SSX－1、－2、－4、－5和NY－ESO－mRNA表达检测。随机抽取每种CTA基因的3例RT－PCR扩增产物的目的片段进行DNA序列测定。结果：35例肺癌标本中MAGE－1、－3，SSX－1、－2、－4、－5和XY－ESO－1的表达率分别为34．3％（12／35）、57．1％（20／35）、17．1％（6／35）、20．0％（7／35）、26．7％（9／35）和37．1％（13／35）。正常肺组织中7种CTA基因均无表达。肿瘤组织中7种基因至少有1种表达的几率是26／35（74．3％），两种或两种以上同时表达几率是23／35（65．7％）。测序结果表明RT－PCR产物确为CTA基因。结论：CTA在肺癌主动免疫治疗中是一种合适的、有前景的攻击靶点。同时，多种CTA的联合表达为多效价CTA疫苗在肺癌免疫治疗中的应用提供了理论基础。     

SSX基因家族与癌-睾丸抗原

SSX(synovial sarcoma X breakpoint,SSX)基因位于X染色体上,是一个由9个成员组成的基因家族。其中一些成员所编码的蛋白质具有癌-睾丸抗原(cancer-testis antigen,CT抗原)的特点,即能在多种肿瘤组织中表达,而在正常组织(除睾丸)几乎不表达。SSX蛋白质具有一定的免疫原性,能在肿瘤患者体内引起体液免疫和细胞免疫。目前的研究结果提示SSX有可能成为用于肿瘤免疫治疗的理想靶抗原。     

MAGE基因mRNA在肺癌细胞中的表达

目的　检测MAGE 1、 2、 3、 4基因在肺癌细胞及癌旁正常肺组织中mRNA的表达水平。方法　用逆转录 -聚合酶链反应 (RT PCR)方法对 3 5例肺癌患者肿瘤组织和癌旁正常肺组织进行MAGE 1、 2、 3、 4mRNA表达检测。结果　 3 5例肺癌肿瘤标本中MAGE 1、 2、 3、 4的阳性表达率分别为 2 2 .9% ( 8/3 5 )、62 .9% ( 2 2 /3 5 )、3 7.1% ( 13 /3 5 )、77.1% ( 2 7/3 5 ) ;四种基因至少有一种表达的几率是 85 .7% ( 3 0 /3 5 ) ;两种或两种以上同时表达几率是 71.4% ( 2 5 /3 5 )。癌旁正常肺组织四种基因均不表达。MAGE基因的表达在肺腺癌和鳞癌 ,在不同分期的非小细胞肺癌间无差异 ,并且与淋巴结转移与否无关 (P >0 .0 5 )。结论　MAGE 1、 2、 3、 4在肺癌组织中有较高比例的表达 ,癌旁正常肺组织中无表达。这提示MAGE抗原用于肺癌的主动免疫治疗具有良好的前景     

SYT-SSX融合基因及其亚型在滑膜肉瘤发生发展中的作用及预后意义

目的研究SYT—SSX融合基因及其亚型（SYT—SSX1和SYT—SSX2）在滑膜肉瘤中的表达情况，探讨它们在滑膜肉瘤发生发展中的作用，并评价其预后意义。方法筛选152例滑膜肉瘤组织标本（来自129例患者），提取石蜡包埋组织中的RNA，采用逆转录-聚合酶链反应（RT—PCR）检测SYT—SSX及其亚型的表达，并比较SYT—SSX1和SYT—SSX2的表达与临床病理参数及预后之间的关系。结果145例（95．4％）组织标本表达SYT—SSX融合基因，其中50例（34．5％）呈SYT—SSX1阳性，91例（62．8％）呈SYT—SSX2阳性，4例（2．8％）SYT—SSX1和SYT—SSX2均为阴性。在SYT—SSX1阳性型滑膜肉瘤中，双相型、细胞角蛋白（CK）阳性和上皮膜抗原（EMA）阳性的比例显著高于SYT—SSX2阳性型（P均〈0．05）。多因素生存分析显示，肿瘤直径≥5cm[相对危险度（RR）=2．325，P=0．027]、有低分化区域（RR=2．122，P=0．048）、出现转移（RR=2．205，P=0．032）和SYT—SSX1阳性（RR=2．775，P=0．008）是提示患者预后不良的独立因素。结论大多数中国滑膜肉瘤患者可能为SYT—SSX2阳性；SYT—SSX亚型可以作为评估滑膜肉瘤患者预后的一项重要因素，SYT—SSX1阳性提示预后不良；SYT—SSX1和SYT—SSX2可能通过对不同的靶基因或对同一些靶基因不同程度的影响，呈现出调控滑膜肉瘤上皮分化能力的差异。     

肝细胞癌中OY-TES-1基因mRNA的表达

 目的：检测OY-TES-1基因mRNA在肝细胞癌(HCC)中的表达水平。方法:建立实时定量PCR方法,检测56例HCC及37例配对癌旁组织中目的基因OY-TES-1及看家基因HPRT的表达,OY-TES -1mRNA的表达量以OY-TES-1/HPRT表示,应用SPSS13.0统计分析软件进行数据分析。结果:（1）经实时定量PCR检测,HCC中OY-TES-1mRNA阳性率为73.21%（41/56）,癌旁组织阳性率为 64.86%（24/37）,有17对HCC及配对癌旁组织均表达阳性,OY-TES-1mRNA在HCC及癌旁组织中 的阳性表达率差异无统计学意义（P＞0.05）;（2）HCC中OY-TES-1mRNA水平明显高于对应的 癌旁组织（P＜0.05）,但与HCC患者的年龄、性别等临床资料无关。结论:OY-TES-1 mRNA在HCC中有较高的表达频率,其表达水平高于对应的癌旁组织,它有望作为用于HCC辅助诊断及免疫治疗的肿瘤抗原。OY-TES-1蛋白在HCC及癌旁组织中的表达及相关意义尚值得做进一步的研究。     

The SSX gene family: characterization of 9 complete genes

Human SSX genes comprise a gene family with 6 known members. SSX1, 2 and 4 have been found to be involved in the t(X;18) translocation characteristically found in all synovial sarcomas. Four (SSX1, 2, 4 and 5) are known to be expressed in a subset of tumors and testis, and anti-SSX antibodies have been found in sera from cancer patients. SSX antigens are thus typical cancer-testis (CT) antigens. To identify additional SSX family members, we isolated and characterized human genomic clones homologous to a prototype SSX cDNA. We also searched public databases for sequences similar to SSX. This identified 3 additional SSX genes, SSX7, 8, 9, and also completed the sequence of the formerly partially defined SSX6 gene. In addition to these novel SSX genes, several SSX pseudogenes were identified. With the exception of 1 pseudogene, all SSX genomic SSX sequences map to chromosome X. Among normal tissues, SSX7 mRNA was present only in testis, whereas SSX6, 8 and 9 were not detected in any normal tissue. SSX6 and 7 were expressed in 1 of 9 melanoma cell lines tested, whereas SSX8 and 9 expression was not detected in any tumor tissue or cell lines tested. SSX1, 2, 4 and 5 mRNA expression can be induced in cell lines by 5-aza-2-deoxycytidine or Trichostatin A. These agents also induce SSX6, but not SSX3, 7, 8 or 9 in the tumor cell lines tested, indicating that mechanisms other than methylation or histone acetylation may be responsible for the repressed state of some SSX genes.     

SSX: A multigene family with several members transcribed in normal testis and human cancer

Analysis of t(X;18) translocation in synovial sarcoma had previously led to the definition of the SSX2 gene, the fusion partner on chromosome X. Subsequent screening of testicular cDNA libraries identified 2 highly homologous genes, SSX1 and SSX3. Among these 3 genes, SSX2 has been found to be identical to HOM-MEL-40, which codes for an immunogenic tumor antigen expressed in various human cancers. SSX2 thus belongs to the family of cancer/testis (CT) antigens, i.e., immunogenic protein antigens with characteristic mRNA expression in normal testis and in cancer. To define additional CT antigens, we have immuno-screened a testicular cDNA expression library with an allogeneic serum from a melanoma patient, and both SSX2 and SSX3 were isolated. Further studies using testicular cDNA and SSX probes defined 2 new members of this gene family, SSX4 and SSX5, while a shorter cDNA variant of SSX4 was also identified. All 5 members of the SSX family shared strong sequence homology, with nucleotide homology ranging from 88 to 95% and amino acid homology ranging from 77 to 91%. Genomic cloning of a prototype SSX gene (SSX2) showed that its coding region is encoded by 6 exons, and the shortened form of SSX4 cDNA represents an alternatively spliced product lacking the 5th exon. Analysis of SSX mRNA expression by gene-specific RT-PCR confirmed that all 5 SSX genes are expressed in testis. In addition, analysis of a panel of 12 melanoma cell lines showed strong mRNA expression of either SSX1 (3/12), SSX2 (3/12), SSX4 (1/12), or SSX5 (1/12), indicating variable activation of the genes in malignant cells. Int. J. Cancer 72:965–971, 1997. © 1997 Wiley-Liss, Inc.     

Heterogeneous expression of GAGE, NY-ESO-1, MAGE-A and SSX proteins in esophageal cancer: Implications for immunotherapy

Cancer/testis antigens (CTAs) elicit immune response in cancer patients and are therefore targets of immunotherapy. Current information on CTA expression is primarily based on mRNA assays and little is known about their expression at the protein level. The objectives of our study are to analyze GAGE, NY-ESO-1, MAGE-A and SSX protein expression in esophageal cancer and to correlate their expression patterns with clinicopathologic parameters and survival. We examined CTA protein expression in 213 patients with esophageal cancer by immunohistochemistry. Antigen-positive tumors were evaluated once and antigen-negative tumors were evaluated 3 times by examining different parts of the cancer specimen. GAGE, NY-ESO-1 and MAGE-A were heterogeneously expressed in 42 (20%), 44 (21%) and 111 (52%) tumors, respectively, whereas SSX expression was not detected. Of the 126 (59%) patients expressing CTAs, 70 (33%) expressed 1, 41 (19%) expressed 2 and 15 (7%) expressed 3 antigens. The expression of MAGE-A was correlated with those of GAGE (p = 0.001) and NY-ESO-1 (p = 0.002), and the expression of GAGE was correlated with that of NY-ESO-1 (p = 0.002). One hundred fifty-six (79%) sections were positively stained in the first evaluation, whereas 37 (19%) and 4 (2%) positive sections were identified in the second and third evaluations, respectively. Particularly, MAGE and GAGE expression showed overlaps. GAGE, NY-ESO-1 and MAGE-A protein expression was not correlated with the disease progression, TNM factors or survival. The detection of immunonegative cells in every specimen suggests addition of other drugs such as 5-aza-2'-deoxycytidine to increase the therapeutic effect of CTA-specific cancer vaccines.