基因芯片技术在食管鳞癌转移相关基因表达谱筛选中的应用

目的 研究人食管鳞癌转移相关基因表达谱,探讨食管鳞癌的转移机制。方法 选取392个与肿瘤转移相关的基因克隆,制备成肿瘤转移基因芯片。提取食管鳞癌组织以及正常食管组织RNA,反转录后标记为cDNA探针,与cDNA芯片杂交,经扫描及Quantarray 3．0软件分析后比较两种组织中的差异表达基因。结果 共筛查出差异表达基因58条,其中表达上调基因36条、下调基因22条,包括癌基因、抑癌基因、黏附分子、基质金属蛋白酶、信号转导因子、细胞代谢和免疫相关基因等。结论 基因芯片筛查食管鳞癌转移相关基因表达谱可为明确食管鳞癌转移机制提供重要参考。     

Claudin-1和VEGF-c在食管鳞癌中的表达

目的检测Claudin-1和VEGF—c蛋白存食管鳞癌组织和癌旁组织中的表达,探讨Claudin-1和VEGF—c蛋白存食管鳞癌发生发展中的作用。方法用组织芯片技术及免疫组化方法检测食管鳞癌及相应癌旁组织中Claudin-1和VEGF-c蛋白的表达,分析其表达与临床病理因素的关系。结果Claudin-1和VEGF—c蛋白在食管鳞癌癌组织中的表达均显著高于癌旁组织。Claudin-1和VEGF—c蛋白在食管鳞癌组织的表达与患者年龄、性刖、肿瘤的分化程度、分级分期和淋巴结转移情况无关。结论食管鳞癌的发生可能与Claudin-1和VEGF-c蛋白的表达上调有关。     

Neuropilin-1在食管鳞癌中的表达及其意义

目的探讨Neuropilin-1(NRP-1)在食管鳞癌中的表达及其生物学和临床意义.方法取42例新鲜原发食管鳞癌和邻近正常食管粘膜标本,所有的标本均经病理诊断确诊.用RT-PCR方法检测每例患者癌组织和相应癌旁正常组织中NRP-1mRNA的表达.癌与癌旁正常组织表达值之间的比较采用配对t检验,癌组织的表达值与临床病理因素之间的关系采用成组t检验,检验结果以P＜0.05为有意义.结果在42例患者中,37例NRP-1mRNA表达强于相应癌旁正常组织,占89.00%,肿瘤直径≥3 cm组、外膜浸润组和淋巴结转移组的NRP-1mRNA表达明显增高.癌组织中NRP-1mRNA的表达与肿瘤分化程度和性别无关.结论在食管鳞癌中NRP-1mRNA的表达明显高于癌旁正常组织,并且与食管鳞癌的发展、浸润和转移有关.     

APRIL mRNA在食管鳞癌组织中的表达及意义

应用SYBR GreenⅠ实时荧光定量PCR法检测63份食管鳞癌、30份正常食管上皮、36份单纯增生、29份轻度不典型增生、10份重度不典型增生、4份原位癌组织中APRIL mRNA的表达,并分析其与临床病理特征的关系.结果 食管鳞癌组织APRIL mRNA表达显著高于正常黏膜组织(P<0.05),且在轻度不典型增生、原位癌及浸润癌中的表达均高于正常黏膜组织(P<0.01或P<0.05).APRIL mRNA的表达水平与淋巴结转移相关(P<0.05).提示APRIL mRNA可作为食管癌早期诊断及预后判断的参考指标.     

RhoC基因mRNA在食管鳞癌中的表达及其意义

目的:探讨RhoC基因在食管鳞癌组织中的表达情况及其与食管鳞癌发生、发展的关系.方法:采用半定量RT-PCR和原位杂交方法检测62例食管鳞癌、31例癌旁不典型增生组织及62例正常食管黏膜组织中RhoC mRNA的相对表达量及细胞定位.结果:食管鳞癌组织中RhoC mRNA表达与癌的组织学分级、浸润深度及淋巴结转移密切相关(P <0.05); 在食管鳞癌癌变过程中,RT-PCR检测RhoC mRNA在癌组织、癌旁不典型增生组织及正常黏膜组织中的表达量依次降低, 分别为0.902±0.119、0.731±0.065、0.653±0.069, 组间比较有明显差异(H= 99.629, P <0.01); 原位杂交检测RhoC转录本主要位于细胞质中, 其在癌组织、癌旁不典型增生组织及正常黏膜组织中的表达率依次降低, 分别为80.6%(50/62)、32.3%(10/31)、21.0%(13/62), 组间比较有明显差异(χ2 =47.735, P <0.01), 两种方法检测的RhoC mRNA表达具有一致性.结论:RhoC mRNA在食管鳞癌组织中显著增加, 并与食管鳞癌生物学行为关系密切, 提示RhoC mRNA过表达与食管鳞癌的发生、发展有关, RhoC mRNA可作为食管鳞癌早期诊断和判断预后的辅助指标.     

C-Jun蛋白在食管鳞癌组织中的表达及临床意义

目的:探讨C-Jun蛋白在食管鳞癌(esophageal squamous cell carcinoma,ESCC)患者组织中的表达及其与临床生物行为之间的关系.方法:运用免疫组织化学及RT-PCR方法检测96例ESCC患者癌组织及相应癌旁织中C-Jun蛋白的表达情况,分析C-Jun蛋白表达于ESCC临床生物学行为的关系.结果:C-Jun在ESCC患者癌组织中的表达(0.7703±0.3330)高于癌旁组织(0.2546±0.1328),差异有统计学意义(t=-11.23,P<0.05),并且C-Jun蛋白在ESCC组织中的表达与肿瘤淋巴转移及TNM分期相关(P<0.05).结论:C-Jun蛋白在ESCC组织中高表达,与肿瘤淋巴转移及TNM分期相关,可能与ESCC的发生发展相关.     

瘦素及其受体在食管鳞癌中的表达意义

目的 探讨瘦素(leptin)及其受体(Ob- Ra、Ob-Rb)与食管鳞癌的关系,为食管鳞癌的发病机制研究提供一定的理论依据.方法 RT- PCR法检测20例正常食管和24例食管鳞癌组织中leptin、Ob-Ra、Ob- Rb的mRNA表达.结果 (1)20例正常食管组织中leptin、Ob- Ra、Ob-Rb的mRNA表达阳性率分别为50.0％、50.0％、40.0％;24例食管鳞癌组织中leptin、Ob- Ra、Ob-Rb的mRNA表达阳性率分别为79.2％、83.3％、83.3％.正常食管组与食管鳞癌组相比差异均有统计学意义(P＜0.05).(2)食管鳞癌组织中leptin、Ob-Ra、Ob-Rb的mRNA相对表达量亦高于正常食管组织,差异均有统计学意义(P＜0.05).结论 瘦素及其受体在食管鳞癌组织中的表达阳性率和相对表达量均高于正常食管组织,提示瘦素及其受体可能在食管鳞癌的发生发展中发挥重要作用.     

SOX2在食管鳞癌组织中的表达及意义

目的:探讨食管鳞癌组织中SOX2 mRNA和蛋白的表达及意义.方法:应用原位杂交和免疫组织化学法分别检测35例正常食管黏膜组织及84例食管鳞癌组织中SOX2 mRNA和蛋白的表达.结果:SOX2在正常黏膜组织中mRNA的表达率为8.6％(3/35),显著低于其在癌组织中的表达率46.4％(39/84),组间比较差异具有...     

miR-130b在食管鳞癌中的表达及对食管鳞癌细胞增殖和迁移的影响

目的:检测微小RNA-130b(miR-130b)在食管鳞癌(esophageal squamous cell carcinoma,ESCC)组织中的表达并探讨其对ESCC细胞增殖、迁移的影响.方法:microRNA(miRNA)芯片筛选ESCC组织中异常表达的miRNAs,TaqMan MGB探针法定量PCR检测19例ESCC组织及配对癌旁组织标本中miR-130b的表达;通过脂质体转染模拟物miR-130b mimics(miR-130bm)促进ESCC细胞Eca109中miR-130b的表达,转染抑制物miR-130b inhibitor(miR-130bi)抑制Eca109细胞中miR-130b的表达;进一步采用CCK-8法和Transwell迁移实验检测ESCC细胞增殖、迁移的变化;生物学信息预测miR-130b的靶基因,双荧光素酶报告基因验证其靶向作用;采用SYBR Green定量PCR和Western blot检测靶基因mRNA和蛋白表达.结果:miR-130b在ESCC组织中的表达明显高于癌旁组织(P<0.01);转染miR-130bm可有效增加ESCC细胞Eca109中miR-130b的表达,进而促进Eca109细胞的增殖和迁移,平均迁移细胞数明显多于对照(112.9±2.4vs54.3±1.8,P<0.01);转染miR-130bi则降低细胞中miR-130b的表达;进而抑制细胞的增殖和迁移,平均迁移细胞数较对照明显减少(33.9±2.3vs56.2±1.9,P<0.01).miR-130b可作用于PTEN3’非翻译区抑制其表达.miR-130b可负向调控PTEN蛋白表达,并促进Akt磷酸化,但对PTENmRNA表达无明显影响.结论:miR-130b在ESCC组织中表达上调,增加其表达促进ESCC细胞Eca109增殖和迁移,降低其表达则抑制Eca109细胞增殖和迁移;miR-130b可在转录后水平负向调控PTEN的表达并促进Akt磷酸化.提示miR-130b有望成为ESCC治疗的新靶点.     

Survivin与COX-2在食管鳞癌中的表达及其相关性研究

目的探讨凋亡抑制基因survivin在食管鳞癌组织中的表达及其与环氧合酶-2(COX-2)的相关性。方法采用免疫组化S-P法检测68例食管癌组织及15例正常食管黏膜组织中survivin蛋白和COX-2蛋白的阳性表达率。结果survivin蛋白在15例正常食管黏膜中呈阴性表达,而68例食管癌组织中45例阳性,占66.1%,差异有显著性。COX-2蛋白在食管癌组织中的阳性表达率为79.4%(54/68)。survivin蛋白阳性率与年龄、性别、原发部位无关,而与分化程度、有无淋巴结转移相关,并与COX-2蛋白阳性表达呈正相关。(Pearson列联系数为0.434)。结论survivin蛋白在食管癌的发生发展中起重要作用,survivin蛋白与COX-2蛋白密切相关,2者可能存在共同的激活机制,构成肿瘤细胞凋亡的多种途径。     

Current understanding of the functional roles of aberrantly expressed microRNAs in esophageal cancer

The incidence of esophageal cancer is rising, mostly because the increasing incidence of esophageal adenocarcinoma in Western countries. Despite improvements in diagnosis and treatment, the overall 5-year survival rates remain low. MicroRNAs (miRNAs) are small non-coding RNA molecules that regulate the expression of target genes. Recently, disease specific miRNAs have been identified, which act as tumor suppressors or oncogenes. In this review, we will summarize the current knowledge about the function of aberrantly expressed miRNAs in esophageal cancer. We selected 5 miRNAs (miRNA-21, -143, -145, -196a and let-7) based on the available literature, and described their potential role in regulating pathways that are deregulated in esophageal cancer. Finally we will highlight the current achievements of using and targeting miRNAs. Because these miRNAs likely have important regulatory roles in cancer development, they open a therapeutic window for new treatment modalities.     

5-Aza-CdR对食管鳞癌甲基化基因的影响

目的探讨5-aza-CdR对食管鳞癌甲基化基因的影响。方法应用人类全基因组寡核苷酸微阵列芯片,荧光双交换法检测5-aza-CdR干预的食管鳞癌细胞Eca-109与正常培养的Eca-109细胞中的差异表达基因,并进行生物信息学分析。结果经统计学分析,共筛选获得384个差异表达基因,其中303个基因表达上调,81个基因表达下调;差异基因的功能涉及信号转导、物质合成代谢、细胞周期、细胞增殖、细胞凋亡、DNA转录、DNA复制、DNA修复、氧化还原、物质运输、免疫反应等;上调表达基因中有138个基因分别含有1～5个CpG岛序列,占总上调基因的45.54%。结论5-aza-CdR可以影响食管鳞癌细胞中基因异常的甲基化修饰,调控肿瘤细胞的异常凋亡和分化,为进一步研究食管鳞癌致病分子机制,提供表观遗传学的依据。     

食管鳞癌组织及周围正常食管黏膜组织差异表达基因的筛选

目的:研究食管鳞癌组织及周围正常食管黏膜组织的差异表达基因,为寻找食管鳞癌早期诊断高敏感性,高特异性的分子指标提供理论依据.方法:分别抽提人食管鳞癌组织及周围正常食管黏膜组织总mRNA,逆转录成cDNA,用单标法以Cy3-dUTP为标记制成探针,与基因芯片进行杂交,筛选出差异表达的基因,并用生物信息学方法做进一步分析....     

MALDI-TOF-MS分析新疆食管鳞状细胞癌的蛋白组学

目的:分析新疆食管鳞状细胞癌和食管正常上皮细胞的差异表达蛋白.方法:运用激光捕获显微切割技术(LCM)分别获取食管鳞状癌细胞和食管正常上皮细胞,应用二维凝胶电泳技术(2-DE)分离纯化细胞,Imagemaster 2D软件比较分析两者电泳图谱的差异,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)鉴定分析两者表达的差异蛋白.结果:建立了食管鳞状癌细胞和食管正常上皮细胞的2-DE图谱,获得43个差异蛋白点,通过质谱鉴定出17种蛋白,其中15种蛋白如Trangelin2、HSP27、S100A11、GSTP等在食管鳞状癌细胞中表达明显增高,2种蛋白如SCCA1,在食管鳞状癌细胞中表达明显降低.结论:提示17种差异蛋白可能与食管鳞状细胞癌的发生和发展有关,为筛选食管鳞状细胞癌的特异性分子标志物奠定基础.     

Epstein-Barr virus association is rare in esophageal squamous cell carcinoma

Background. A critical role of Epstein-Barr virus (EBV) in carcinogenesis of nasopharyngeal squamous cell carcinoma and gastric adenocarcinoma is strongly suspected. We analyzed the possible EBV association for Japanese squamous cell carcinoma (SCC)-dominant esophageal cancer cases. Methods. We retrospectively screened 36 surgically resected esophageal cancer lesions from 36 patients maily with SCC using in situ hybridization (ISH) for EBV-encoded small RNA1 (EBER-1). EBV DNA analysis using real-time quantitative polymerase chain reaction (Q-PCR) was performed for three recent cases. Results. We found no EBER-1-positive cancer cell in any tested esophageal cancer lesion. There were many EBER-1-positive tumor-infiltrating lymphocytes in the basaloid SCC lesion and a small number of positive lymphocytes in the other five advanced SCC lesions (14.7% of SCC). One SCC lesion with a highcopy number of EBV DNA had EBER-1-positive lymphocytes. Conclusions. EBV is rarely associated with esophageal SCC, and may appear through tumor-infiltrating lymphocytes in some advanced lesions.     

Human papillomavirus in esophageal squamous cell carcinoma in Colombia and Chile

INTRODUCTION Human papillomavirus (HPV), a double-stranded DNA virus, is recognized as an etiologic agent of cervical cancer[1]. In addition, HPV is suspected of causing extragenital cancers, including cancers of the oral cavity, larynx, esophagus, and lu…     

Endoscopic diagnosis and treatment of esophageal verrucous squamous cell cancer

Verrucous squamous cell cancer (VSCC) of the esophagus is a variant of squamous cell carcinoma. This rare entity has been described in only a handful of case reports in the literature. We sought to evaluate the endoscopic features, treatment, and outcomes related to esophageal VSCC. The medical records of all patients with esophageal VSCC seen at our institution from January 1995 to December 2010 were reviewed retrospectively. A total of 11 patients (6 men; mean age 66 years [range 57–75 years]) were identified, with a mean follow up of 4 years (range 0.5–10 years) available in nine patients after diagnosis. About half the patients smoked or consumed alcohol on a regular basis. The median time interval from onset of symptoms to diagnosis of esophageal VSCC was 2.5 years (range 1–20 years), with dysphagia being present in all patients. The majority of tumors (8 of 11) exhibited a white, warty, plaque‐like appearance with superimposed Candida at endoscopy, which led solely to a diagnosis of Candida esophagitis on initial presentation. The disease was either extensive (n = 5) throughout the esophagus or localized (n = 6) often by tumor nodules or projections, with the lower third of the esophagus being most commonly involved. Initial pinch biopsies were nondiagnostic in eight (73%) of the patients. Six patients underwent esophagectomy; neoadjuvant chemoradiation therapy was provided in two. In patients treated solely with surgery and who had a preoperative endoscopic ultrasound, the latter tended to overestimate staging of the lesion relative to surgical pathologic staging. Two patients were deemed to be poor operative candidates and received only chemoradiation treatment. One patient with a T2N0 tumor by endoscopic ultrasound staging was managed symptomatically with intermittent endoscopic dilation because of significant comorbidities that precluded surgery and oncologic therapy. There has been no evidence for residual or recurrent neoplastic disease in the eight patients who received treatment with surgery and/or chemoradiation therapy. Five of six patients who underwent surgery have required intermittent endoscopic dilation of anastomotic strictures during follow up. One of the two patients who received only chemoradiation therapy has required periodic endoscopic dilation for radiation‐induced esophageal stricture. Two of the nine (22%) patients have died of causes unrelated to VSCC or its treatment at last follow up. In conclusion, a high index of suspicion for esophageal VSCC should be raised by the presence of long‐standing symptoms coupled with white, warty esophageal lesions seen on endoscopic evaluation. Candida overgrowth can be expected to confound the diagnosis. Despite the long duration of symptoms, surgical resection typically shows relatively low‐grade tumors, consistent with the rare propensity of this variant of esophageal squamous cell carcinoma to metastasize.