The MAGE-A1 gene expression is not determined solely by methylation status of the promoter region in hematological malignancies

Tumor antigens such as MAGE-A1 are aberrantly expressed in many human tumors and could be recognized by CTL. Thus, they could be targets for cancer immunotherapy. It is presently considered that the expression of the MAGE-A1 gene is regulated by methylation of its promoter region. To estimate the possibility of activating the MAGE-A1 gene with demethylating agents with a view toward clinical use, we assessed the methylation status of its CpG-rich promoter by sodium bisulfite mapping both of samples that express the gene and those that do not. Cell lines and samples from patients with hematological malignancies were examined. Surprisingly, the methylation status of the MAGE-A1 gene did not clearly correlate with the expression of the gene. Our results indicate that the MAGE-A1 gene expression is not determined solely by the methylation status of the promoter region in hematological malignancies.     

肺腺癌癌旁组织及胚胎肺的基因表达谱研究

目的 利用基因芯片技术研究肺腺癌组织、癌旁组织及胚胎肺组织中的基因表达差异。方法 分别抽取肺腺癌组织、癌旁组织和胚胎肺组织的总RNA,并纯化mRNA。采用逆转录的方法,制成cDNA链,并以两种荧光Cy5和Cy3标记后做探针,与含有1152条人类全长基因芯片进行杂交。以ScanArray 4000荧光扫描仪扫描芯片上两种荧光信号,并用计算机处理和分析。结果 4例肺腺癌组织和癌旁组织标本中,共同表达差异基因25个,其中上调基因3个,下调基因22个;胚胎肺组织和癌旁组织标本中,表达差异基因316个,其中上调基因192个,下调基因124个;胚胎肺组织和癌旁组织中表达差异基因与肺腺癌组织和癌旁组织比较,共同表达差异基因16个,其中上调基因12个,下调基因4个。结论 肺腺癌与癌旁组织共同表达差异的25个基因可能与肺癌的发生、发展有关;胚胎肺组织与癌旁组织表达差异的316个基因与生长发育环境有关;胚胎肺组织和癌旁组织与肺腺癌组织和癌旁组织共表达差异的16个基因可能与早期肺腺癌启动有关。     

Genes associated with early development, apoptosis and cell cycle regulation define a gene expression profile of adenoid cystic carcinoma

Adenoid cystic carcinoma (ACC) is an uncommon salivary gland malignancy characterized by indolent yet relentless growth that exhibits inherent resistance to systemic chemotherapy, surgical salvage and conventional radiotherapy. We used microarray analysis to characterize gene expression changes associated with ACC. Eight ACC patient specimens were compared with normal parotid gland tissue and the ACC3 cell line. Differentially expressed genes were identified (512 total) using supervised analysis methods and functional categories assigned using OntoExpress. Genes associated with morphogenesis, neurogenesis, proliferation and apoptosis characterized ACC tumors. Genes associated with saliva production and immune response characterized normal parotid tissues while the ACC3 cell line expressed genes primarily associated with proliferation, chromosome maintenance and the cell cycle. These results demonstrate that ACC tumors express genes associated with early developmental processes including morphogenesis and neurogenesis implicating oncogenic events that result in dedifferentiation of normal salivary glands.     

Prognostication and prediction using gene expression profiling in oesophageal cancer

Aim

To evaluate current literature on gene expression profiling in oesophageal cancer.

Methods

We performed a review of the literature (2000–2010) on prognostication and prediction using gene expression analysis in oesophageal cancer.

Results

Seventeen papers comprising 638 patients were included. Gene expression profiles studied in relation to survival, lymph node metastasis and response to neoadjuvant therapy. Most studies included a limited number of patients. Several prognostic and predictive gene signatures were identified with different accuracies. In only one study, the gene signature was validated in a large, independent patient cohort.

Conclusion

Gene expression profiling has potential clinical applications in oesophageal cancer. Especially a signature which is predictive for response to neoadjuvant treatment could be of great clinical value. To date, most published studies suffer from an underpowered training cohort or lack adequate validation. Clinicians should put effort in the collection of high quality tissue samples and should participate in biobank initiatives, considering the increasing availability and possibilities of sequencing technology.     

人睾丸肿瘤抗原MAGE-E1基因的克隆测序及在原核系统中的表达

目的：扩增及克隆人的MAGE-E1基因并利用大肠杆菌进行表达。方法：从人胶质瘤细胞系BT-325中提取总RNA，用RT-PCR从中扩增出MAGE-E1基因片段。将MAGE-E1基因片段插入载体pGEM-T easy中，经全自动序列分析仪测序正确后，再克隆至表达载体pGEX-4T-2中，构建重组表达载体pGEX-4T-2-MAGE-E1，并转化大肠杆菌进行表达。结果：1mmol／L．异丙基-β-D-硫代半乳糖苷(IPTG)诱导5h，MAGE-E1蛋白表达即达高峰，SDS-PAGE及凝胶密度扫描分析表明，表达出Mr约41000大小的蛋白，占菌体总蛋白的35％。结论：成功扩增、克隆MAGE-E1基因，并在大肠杆菌中得到稳定高效表达。     

一种新型肿瘤组织起源分子标志物的建立与评价

背景与目的：原发灶不明恶性肿瘤是一类转移性肿瘤的统称,在诊断时无法找到原发位点,约占所有恶性肿瘤的5%~10%。明确肿瘤的组织起源对于患者的诊断和治疗具有重要意义。方法：整合ArrayEx-press和Gene Expression Omnibus数据库中肿瘤类型明确的样本数据,构建涵盖22种常见肿瘤类型、5800例样本的基因表达谱数据库;通过支持向量机递归特征消除算法筛选组织特异性基因,建立肿瘤分类模型;采用实时定量聚合酶链反应(real-time quantitative polymerase chain reaction,RTQ-PCR)检测石蜡包埋肿瘤组织中基因的表达水平,并将基因分型结果与病理诊断结果进行比较。结果：基于肿瘤基因表达谱大数据,筛选出96个组织特异性基因,其中包含常见的肿瘤相关基因,如钙黏蛋白1(cadherin 1,CDH1)、激肽释放酶相关酶3(kallikrein-related peptidase 3,KLK3)和表皮生长因子受体(epidermal growth factor receptor,EGFR)等。在206例石蜡包埋组织样本中,182例的基因分型结果与病理诊断结果一致,准确率达到88.4%(95%CI：83.2%~92.4%)。结论：96基因RTQ-PCR检测对22种常见肿瘤类型具有较好的分类性能,可作为临床和病理诊断的辅助工具。     

新基因pp3774的亚细胞定位以及在肝癌细胞系和组织中的表达谱研究

目的探讨新基因pp3774在肝癌细胞系、人肝癌及癌旁肝、肝硬化以及正常肝组织中的表达差异及其生物学功能.方法脂质体转染-荧光观察pp3774-GFP融合蛋白的亚细胞定位;合成新基因pp3774特异性多肽,制备兔源性多克隆抗体;用RT-PCR、蛋白印迹、免疫细胞化学及免疫组化方法检测pp3774基因在7个肝癌细胞系及214例肝细胞癌、18例肝硬化及10例正常肝组织中的表达差异.流式细胞术检测转染pp3774基因的SMMC-7721细胞的凋亡率.结果 pp3774蛋白主要定位于细胞质(内质网为主);RT-PCR检测结果表明pp3774 mRNA在肝癌细胞系BEL-7402、Huh-7、MHCC-97L及HepG2中高表达,SMMC-7721、MHCC-LM3及人永生化肝细胞系L-02中度表达,而在Hep3B中低表达;肝癌及癌旁组织中pp3774 mRNA的表达表现为癌旁高于癌(P<0.05);免疫组化结果显示pp3774蛋白表达程度依次为正常肝≥肝硬化≥癌旁肝组织>肝癌.转染pp3774基因的SMMC-7721细胞的凋亡发生率高于对照组约10%.结论 pp3774基因是一个具有抑制肝癌细胞生长功能的新基因.     

利用组织微阵列研究RASSF1A和Survivin基因在非小细胞肺癌中的表达

目的探讨RASSF1A和Survivin基因的蛋白表达与非小细胞肺癌（NSCLC）临床病理特征的关系及其临床意义。方法免疫组织化学法检测RASSF1A和Survivin在NSCLC组织微阵列中的蛋白表达。结果RASSF1A蛋白在NSCLC中的阳性率（46．8％〉显著低于正常肺组织（92．9％）（P〈0．001），但Survivin阳性率（75．8％）显著高于正常肺组织（0）（P〈0．001）；RASSF1A蛋白在临床Ⅰ期和Ⅱ期NSCLC中分别显著高于临床Ⅲ期（P〈0．001，P〈0．001），Survivin在临床I期和临床Ⅱ期NSCLC中的阳性率显著低于临床Ⅲ期者（P=0．003，P=0．001），淋巴结转移性NSCLC的RASSF1A阳性率显著低于无淋巴结转移者（P〈0．05）；RASSF1sA和Survivin蛋白在NSCLC中的表达呈负相关（r=-0．780，P〈0．001）。结论RASSFlA蛋白表达下调、Survivin蛋白高表达及其两者的表达失平衡在NSCLC的发生、发展中可能具有重要作用，RASSF1和Survivin有望成为评估肺癌淋巴结转移和预后预测的重要分子标志。     

肺癌中ING1基因变化及其表达水平的研究

目的：检测肺癌中抑癌基因ING1微卫星杂合性缺失（LOH）及其主要蛋白产物p33^ING1b的表达情况，以探讨ING1基因改变-9肺癌发生发展的关系。方法：采用银染PCR—SSCP法检测肺癌组织ING1基因微卫星LOH发生的频率；应用组织芯片技术和免疫组化方法，检测肺癌p33^ING1b蛋白表达水平。结果：70例肺癌组织ING1基因4个微卫星位点的总杂合性缺失率为55．7％（39／70），并且越靠近ING1基因位点，发生率越高，但与临床病理参数无关。217例肺癌p33^ING1b蛋白的LOH发生率为47．0％（102／217），鳞状细胞癌高于腺癌、腺鳞癌和细支气管肺泡癌（P〈0．05），其余类型间差异无显著性（P〉0．05）；p33^ING1b的表达与其他临床病理参数不相关（P〉0．05），但与LOH发生频率相关（P〈0．05）。结论：p33^ING1b蛋白在肺癌组织中存在高频率的低表达或失表达，并且ING1基因的微卫星LOH发生频率也很高。推测LOH是该基因异常表达的重要原因，其结果可能导致该基因的下调和（或）蛋白质功能的失活，从而丧失其对细胞的生长抑制作用，促进了肿瘤的发生。     

RNA 结合因子 AUF1调控基因表达及其在肿瘤发生中的双重调控作用的研究进展

信使RNA（ Messenger ribonucleic acid ,mRNA）不断更新在基因表达中具有重要作用,而且编码一些重要蛋白。本文详细阐述了RNA结合因子1（RNA-binding factor 1,AUF1）使mRNA稳定或去稳定,在转录后水平调控基因表达的可能的分子机制,归纳了AUF1各个亚型蛋白在基因表达中的独特作用的相关研究进展,并探讨了AUF1表达水平的改变在肿瘤发生中的双重调控作用。认识到AUF1发挥生物学效应的分子机制以及作用的具体信号通路,能够帮助我们揭开更多的生命谜题。     

Constitutive expression and activation of stress response genes in cancer stem-like cells/tumour initiating cells: Potent targets for cancer stem cell therapy

Abstract

Cancer stem-like cells (CSCs)/tumour-initiating cells (TICs) are defined as the small population of cancer cells that have stem cell-like phenotypes and high capacity for tumour initiation. These cells may have a huge impact in the field of cancer therapy since they are extremely resistant to standard chemoradiotherapy and thus are likely to be responsible for disease recurrence after therapy. Therefore, extensive efforts are being made to elucidate the pathological and molecular properties of CSCs/TICs and, with this information, to establish efficient anti-CSC/TIC targeting therapies. This review considers recent findings on stress response genes that are preferentially expressed in CSCs/TICs and their roles in tumour-promoting properties. Implications for a novel therapeutic strategy targeting CSCs/TICs are also discussed.     

乳腺癌组织中NOEY2基因的mRNA表达及其与临床病理的关系

目的：观察乳腺癌组织中NOEY2基因的mRNA表达情况,并探讨其与临床病理及其他分子指标的关系。方法：以RT－PCR法和反义RNA探针原位杂交法,检测乳腺良恶性病变组织中NOEY2基因的mRNA表达,以免疫组化法检测乳腺癌石蜡标本中的雌激素受体（ER)状态,Ki67标记指数以及p27和p21WAF1的表达。结果：RT－PCR法检测显示,24例冻存乳腺组织中,6例良性病变NOEY2均为阳性;18例乳腺癌中,13例（72．2％）NOEY2为阳性,原位杂交法检测显示,10例乳腺良性病变NOEY2均为阳性;而60例乳腺侵袭性癌中,仅有31例NOEY2为阳性,占51．7％,良恶性病变NOEY2阳性率差异有显著性（P＜0．025）,NOEY2的阳性率有随组织学分级升高而递减的趋势。淋巴结阴性者阳性率为76．7％,而阳性者仅为26．7％,两者差异有显著性（P＜0．001）,NOEY2基因mRNA表达与其他临床病理指标以及免疫组化指标均无显著相关性。结论：NOEY2基因可能在一定程度上参与乳腺癌的发生和发展。该基因的失表达可能与乳腺癌的转移机制有关。     

EXPRESSION OF mdr-1 GENE IN CANCER TISSUE AND ITS ASSOCIATION WITH MORPHOLOGICAL INDEXES OF ESOPHAGEAL CARCINOMA IN ANYANG

Multipledrugresistance(MDR)ofcancercellsisoneofthemostinterestingareasinthecurrentcancerresearches.IthasbeenshownthatmanytumorshaveMDR.OneofthemolecularbaseofMDRistheamplificationofmdr-lgeneandoverexpressionofitsproduct,pl70,whichwerethoughtasthedirectcauseofchemothcrapyfailurebymanyinvestigators.'--'Moreover,othersicbelievethatmdr-1geneexpressionincancertissuewasamalignantbiologicalindicatorforneoplasms.UPtodate,fewresearchreportswerefoundintheliteratureonmdr-lgeneexpressioninesophaguscan…     

ERCC5/XPG, ERCC1, and BRCA1 gene status and clinical benefit of trabectedin in patients with soft tissue sarcoma

BACKGROUND:

The objective of this study was to determine whether specific single nucleotide polymorphisms (SNPs) from nucleotide excision repair (NER) and homologous recombination (HR) DNA repair pathways are associated with sensitivity to trabectedin in patients with soft tissue sarcoma (STS).

METHODS:

The authors analyzed excision repair cross‐complementation group 5/xeroderma pigmentosum group G (ERCC5/XPG) (NER), excision repair cross‐complementation group 1 (ERCC1) (NER), and breast cancer 1 (BRCA1) (HR) SNPs and messenger RNA expression levels in tumor specimens from 113 patients with advanced STS who were enrolled in previously published phase 2 trials or in a compassionate‐use program. The 6‐month progression‐free rate (PFR), progression‐free survival (PFS), and overall survival (OS) were analyzed according to ERCC5, ERCC1, and BRCA1 status using log‐rank tests.

RESULTS:

High expression of the common allele (aspartic acid at codon 1104) of ERCC5, high expression of ERCC1, and BRCA1 haplotype were associated significantly with improved PFR, PFS, and OS. The ERCC1 thymine‐to‐cytosine (T→C) SNP at codon 19007 and BRCA1 expression were not associated with outcome. On univariate analysis, tumor histology, favorable NER status (high expression of common ERCC5 and/or high ERCC1 expression status), and favorable BRCA1 haplotype (at least 1 triple‐adenine plus guanine [AAAG] allele) were the sole variables associated significantly with PFS and OS.

CONCLUSIONS:

In the current study, ERCC5, ERCC1, and BRCA1 status represented a potential DNA repair signature that could be used for the prediction of clinical response to trabectedin in patients with STS. Cancer 2011. © 2011 American Cancer Society.     

食管癌中MMP-9、uPA和uPAR蛋白表达及其临床意义

目的 探讨食管癌中MMP-9、uPA和uPAR蛋白表达与临床病理因素间的关系及其对单纯放疗预后的影响.方法 应用免疫组化SP法检测59例食管鳞癌和41例癌旁组织中MMP-9、uPA和uPAR蛋白表达情况,分析蛋白表达与临床病理因素间的关系及其对食管癌放疗预后的影响.结果 59例食管癌组织中MMP-9、uPA、uPAR蛋白阳性表达率分别为85%、76%、78%,41例食管癌旁组织中阳性表达率分别为39%、49%、44%,癌组织和癌旁组织中3种蛋白的阳性表达率差异均有统计学意义(χ2=22.54、8.04、12.18,P值均＜0.01).胸部CT扫描显示食管病变外侵深度＞2 am者MMP-9蛋白阳性表达率为100%,明显高于外浸深度≤2 cm的79%(P=0.048).胸部CT扫描显示食管病变层面椎前三角形间隙消失者uPA蛋白强阳性表达率为70%,明显高于三角形间隙存在者的44%(χ2=4.21,P=0.040).发生远处转移者14例,其uPA蛋白阳性表达率较无远处转移者也显著升高,分别为100%和69%(χ2=4.12,P=0.042).进一步分析发现MMP-9、uPA、uPAR蛋白表达对食管癌放疗后的近期生存均未见明显影响,而MMP-9与uPA、MMP-9与uPAR、uPA与uPAR蛋白表达间均呈显著正相关.结论 食管癌中MMP-9、uPA蛋白高表达与肿瘤局部外侵及远处转移有关,可能促进了肿瘤局部浸润及远处转移的发生.3种蛋白表达对食管癌放疗预后的影响有待长期随访和大宗病例的研究.     

切除修复交叉互补基因1表达与非小细胞肺癌患者吉西他滨联合顺铂化疗疗效的相关性

目的探讨非小细胞肺癌(NSCLC)患者肿瘤组织内核苷酸切除修复交叉互补基因1(ERCC1)表达与吉西他滨联合顺铂(GP方案)化疗疗效的相关性。方法采用免疫组化S-P法检测51例Ⅱb～Ⅳ期NSCLC患者肿瘤组织内ERCC1的表达。所有患者接受至少2个周期以上的GP方案化疗,对患者化疗后的中位生存期(MST)及化疗后反应率(RR)进行评估。结果 51例患者ERCC1的阳性表达率为43.14%(22/51)。ERCC1表达阳性组化疗后的RR和MST分别为50.0%、17个月,与阴性组RR(31.04%)、MST(12个月)比较,差异无统计学意义(χ2=1.8877,P=0.1695;χ2=1.6767,P=0.1954)。结论 ERCC1表达阳性的NSCLC患者能从吉西他滨联合顺铂的化疗中受益,调控ERCC1表达逆转耐药的策略,将为肿瘤治疗带来新的方法和途径。