食管癌发生发展过程中p53蛋白表达的研究

目的研究p53蛋白在食管癌及癌前病变组织中的表达,探讨其在食管癌发生、发展过程中的作用。方法应用免疫组织化学方法检测43例鳞状细胞癌及22例增生性病变(单纯性增生8例,轻、中、重度不典型增生分别为6例、4例、4例)和6例正常食管黏膜鳞状上皮的食管组织中p53蛋白的表达情况。结果在正常食管黏膜上皮,单纯性增生,轻、中度不典型增生中,未发现p53蛋白的表达;在75%(3/4)重度不典型增生、56%(24/43)食管鳞状细胞癌中p53蛋白表达阳性。p53蛋白表达在I～II期为18%(2/11),III～IV期为77%(10/13)。p53蛋白表达,与淋巴转移、浸润深度、肿物大小无关。结论 p53蛋白的表达与食管癌发生发展关系密切,是食管癌发生发展中的一个重要指标。可作为食管癌可疑病例定期随访的指标之一,以便发现早期食管癌,做到早诊断早治疗。     

卢戈液染色与p53蛋白检测诊断早期食管癌的可行性研究

目的　验证内镜下卢戈液染色和活组织p5 3检测 ,诊断食管早期癌和表浅癌的可行性。方法　将 97例食管癌手术标本先用卢戈液染色 ,然后对粘膜不着色区行p5 3蛋白免疫组化染色。结果　共有 6 3处直径大于 2mm的粘膜不着色区 ,其中正常粘膜 2处 ,单纯增生 7处 ,不典型增生 42处 ,原位癌 1处 ,鳞状细胞癌 11处 ;p5 3蛋白的阳性表达主要发生在重度不典型 (42 3% )和鳞状细胞癌 (45 5 % )的细胞核内 ,而在正常粘膜、单纯增生和轻至中度不典型增生组织中几乎无表达 (P <0 0 5 )。结论　内镜下卢戈液染色和活组织p5 3检测诊断食管早期癌和表浅癌的设想具有可行性。     

水通道蛋白4在食管鳞癌中的表达

目的 观察水通道蛋白4(AQP4)在食管鳞癌组织中的表达并探讨其在食管癌发病中的作用.方法 取食管鳞癌组织、癌旁正常鳞状上皮组织各16例,应用免疫组织化学,逆转录.聚合酶链反应(RT-PCR)技术检测AQP4的表达及分布.结果 免疫组织化学显示,AQP4表达于正常食管黏膜鳞状上皮细胞,在食管鳞癌组中主要表达于肿瘤上皮细胞和癌巢中.RT-PCR法结果 显示,AQP4在癌旁正常组织和食管癌组织中的mRNA表达平均相对A值分别为0.45±0.12、0.70±0.23,差异有统计学意义(P＜0.05).结论 AQP4在正常食管黏膜鳞状上皮细胞以及食管鳞癌组中均有表达,而且在癌组织中表达增加;AQP4可能对人食管癌的发生、发展起促进作用.     

食管癌早期诊断对手术方式的影响

目的探讨食管癌早期诊断对手术方式的影响。方法对3 000名年龄41⁶9岁的农村健康成年人人群进行早期食管癌筛查。行胃镜检查后,内镜退至食管时进行2%卢氏碘染色,并对浅染色和不染色的黏膜进行活检,送病理检查。结果在3000人被检查对象中,查出食管黏膜轻度不典型增生700例,中度不典型增生6例。筛查出需要治疗的阳性患者共9例,其中食管黏膜中度不典型增生1例,食管黏膜高级别上皮内瘤变3例,食管黏膜鳞状细胞癌5例,其中,有3例黏膜病变<2 cm,2例黏膜病变>2 cm。在9阳性患者中,4例患者已经在上级医院行内镜下手术治疗。结论筛查并诊断出早期食管癌患者,可以选择内镜下手术治疗,减轻患者痛苦及经济负担,提高生存时间。     

内镜黏膜下隧道法切除早期食管癌及癌前病变的应用分析

目的 探讨内镜黏膜下隧道法在切除早期食管癌及癌前病变中的应用价值.方法 17例术前超声内镜检查判断病变局限于黏膜层,经黏膜活检发现食管上皮局灶癌变或重度不典型增生的患者,采用黏膜下隧道法的内镜下早期癌切除.结果 17例中,术后病理确诊鳞状上皮增生伴黏膜慢性炎4例,重度不典型增生5例,高至中分化鳞癌8例,其中T1a期7例,T1b期1例.有2例切除黏膜边缘重度不典型增生,1例黏膜下层切缘见癌细胞,其余病例均病灶完整切除.术后1例患者因迟发性出血转开胸手术治疗,其余患者均恢复良好.结论 黏膜下隧道法切除黏膜内早期食管癌及癌前病变安全、有效,更符合直视、充分暴露的外科原则,明显减少出血、穿孔的并发症风险,但其对病灶切除范围判断有一定困难,需在手术中充分注意.     

Src家族酪氨酸激酶Fyn在人食管鳞状细胞癌中的表达

目的 探讨食管鳞状细胞癌组织中Fyn的表达水平及其意义.方法 收集13例食管鳞状细胞癌组织(ESCC)和10例食管正常黏膜上皮组织(UNR),采用免疫组织化学法和免疫印迹法分析食管鳞状细胞癌组织及食管正常黏膜组织Fyn蛋白表达水平的变化.结果 免疫组织化学染色显示食管鳞状细胞癌组织中Fyn蛋白表达水平(9.68±2.31)高于食管正常黏膜组织中染色指数(3.21±1.25),差异有统计学意义(P<0.01).免疫印迹结果亦显示食管鳞癌组织中Fyn蛋白表达强于食管正常黏膜组织.结论 Fyn蛋白在食管鳞状细胞癌组织中高表达,提示Fyn基因在食管鳞状细胞癌的发生发展过程中可能起重要作用.     

食管鳞状细胞癌组织中酪氨酸激酶受体B和脑源性神经生长因子蛋白及mRNA的表达及意义

目的 探讨酪氨酸激酶受体B( TrkB)和脑源性神经生长因子(BDNF)蛋白及mRNA在食管鳞状细胞癌(ESCC)组织中的表达及其意义.方法 应用免疫组织化学及原位杂交法检测59例ESCC、27例癌旁不典型增生组织及36例正常食管黏膜组织中TrkB和BDNF蛋白及mRNA的表达.结果 ESCC组织中TrkB蛋白及mRNA的阳性表达率分别为71.2％和64.4％,显著高于癌旁不典型增生组织(阳性率分别为48.1％和33.3％)及正常食管黏膜组织(阳性率均为0.0％),组间比较差异有统计学意义(P＜0.05);此外,BDNF蛋白和mRNA在ESCC组织中的阳性表达率分别为76.3％和69.5％,也显著高于癌旁不典型增生组织(阳性率分别为55.6％和40.7％)及正常食管黏膜组织(阳性率均为0.0％),组间比较差异有统计学意义(P＜0.05).TrkB和BDNF蛋白及mRNA的表达均与ESCC的分化程度、浸润深度和淋巴结转移密切相关(P＜0.05).进一步相关分析结果显示,TrkB mRNA和蛋白的表达均与BDNF mRNA和蛋白的表达呈正相关(P＜0.05).结论 TrkB和BDNF蛋白及mRNA表达与ESCC的发生、发展和转移密切相关.     

EGFR基因蛋白在食管癌组织的表达及临床意义

目的　探讨EGFR基因蛋白在食管癌中的表达与食管癌的发生发展及浸润转移之间的关系。方法　采用免疫组织化学方法检测 58例食管癌标本中和 12 2例食管上皮其他各组病变中EGFR的表达水平及EGFR的表达与食管癌的临床病理特征关系。结果　正常组 ( 0 2 0 ,0 % )、轻度不典型增生 ( 6 32 ,18.8% )及中度不典型增生组 ( 11 19,57.9% )相邻组间EGFR表达均有显著性差异 (P<0 .0 5) ;Ⅰ级鳞癌 ( 14 2 2 ,6 3.6 % )与中 ( 11 19,57.9% )、重度不典型增生 ( 9 15,6 0 .0 % )、Barrett’s食管( 5 8,6 2 .5% )组间EGFR表达均无显著性差异 (P >0 .0 5)。高、中分化组 ( 16 39,4 1.0 % )与低分化组( 11 19,57.9% )间EGFR表达差异无显著性 (P =0 .10 86 ) ;有淋巴结转移者 ( 2 3 39,59.0 % )与无淋巴结转移者 ( 4 19,2 1.1% )相比较 ,差异有显著性 (P =0 .0 0 56 )。结论　随着肿瘤恶性程度及浸润转移的增加 ,EGFR基因蛋白表达逐渐增强 ;EGFR可作为监测肿瘤早期发生 ,预测转移潜能和预后状况的有价值的指标 ;并以此提出新的治疗策略     

食管鳞状细胞癌及癌前病变组织中多个染色体区域微卫星杂合性缺失的研究

目的 探讨与食管癌早期癌变的发生、发展密切相关的分子变化,寻找早期诊断的分子标记.方法 采用PCR荧光测序仪凝胶电泳分别检测34例食管癌患者手术切除的鳞状细胞癌(SCC)组织、癌前病变组织及匹配的外周血样本中16个微卫星位标的杂合性缺失状况.将轻、中度不典型增生定义为低级别不典型增生(LGD),将重度不典型增生归为高级别不典型增生(HGD).比较不同病理形态组织中16个位标的杂合缺失率.结果 16个位标的总体缺失率随组织形态的严重程度而升高,LGD中缺失率(9.8%)低于HGD(48.6%)和SCC(58.5%,P＜0.01).有8个位标,即D3S1597、D3S2452、D3S1285、D4S174、D5S2501、D9S125、D13S153和D17S786,在LGD中已呈现杂合性缺失;另发现4例患者的HGD和SCC样本分别在D3S2452、D4S174、D9S125及D17S261处出现了杂合性缺失的逆转.结论 食管鳞状上皮从不典型增生至癌变的过程中需要一系列分子变化的积累;针对某些位标的杂合性缺失分析将有助于食管癌的早期诊断;食管癌的发生可能存在遗传异质性.     

TPX2在食管胃交界部腺癌中的表达及临床意义

目的研究TPX2在食管胃交界部腺癌组织中的表达,并探讨其临床意义。方法采用免疫组织化学法分析80例食管胃交界部腺癌、41例癌旁不典型增生组织及48例正常胃黏膜组织中TPX2蛋白的表达;运用RT-PCR法检测TPX2mRNA在40例食管胃交界部腺癌、36例癌旁不典型增生组织和28例正常胃黏膜组织中的表达水平。结果食管胃交界部腺癌、癌旁不典型增生组织及正常胃黏膜组织中的TPX2蛋白阳性率分别为71.2%(57/80)、53.7%(22/41)和31.2%(15/48),三组间比较差异有统计学意义(χ~2=10.315,P=0.018);TPX2 mRNA阳性率分别为77.5%(31/40)、69.4%(25/36)和7.1%(2/28),三组间比较差异也有统计学意义(χ~2=7.468,P=0.025)。TPX2蛋白及mRNA在食管胃交界部腺癌组织中表达的阳性率随着肿瘤浸润深度增加、淋巴结转移和肿瘤分期增高而逐渐增高(P0.05),而与性别、年龄及组织分化程度均无关(P0.05)。结论 TPX2的异常表达可能是食管胃交界部上皮细胞癌变与浸润转移的重要因素,TPX2的检测有利于食管胃交界部腺癌的早期诊断和预后判断。     

Molecular determinants in targeted therapy for esophageal adenocarcinoma

HYPOTHESIS: Cyclooxygenase 2 (COX-2), vascular endothelial growth factor (VEGF), and epidermal growth factor receptor (EGFR) are useful biological determinants in targeted therapy for esophageal adenocarcinoma. DESIGN: Prospective analysis. SETTING: University tertiary referral center. PATIENTS: Sixteen patients with squamous mucosa and normal results of a pH study without mucosal injury (control group), 15 with Barrett esophagus (metaplasia group), and 44 with adenocarcinoma (carcinoma group). INTERVENTIONS: Biopsy specimens were obtained 3 cm above the gastroesophageal junction. Dysplastic tissue was additionally isolated from 9 of the patients in the carcinoma group. After laser-capture microdissection, quantitative real-time polymerase chain reaction was used to measure gene expression across the spectrum of the metaplasia-dysplasia-carcinoma sequence. MAIN OUTCOME MEASURES: Expression of COX-2, VEGF, and EGFR in each patient group. RESULTS: Expression of both COX-2 and VEGF was significantly up-regulated in patients with metaplasia, dysplasia, and cancer compared with controls (P<.01). Expression levels of both were significantly higher in cancer than in the metaplasia group (P<.05) and increased sequentially from metaplasia to dysplasia to cancer. Expression of VEGF was significantly higher in the dysplastic tissue than in nondysplastic Barrett epithelium (P<.05). No change in expression levels of EGFR was seen in the histologic progression to esophageal adenocarcinoma. CONCLUSION: Gene expression data suggest that pharmacologic inhibition of COX-2 and VEGF may be useful adjuncts in targeted therapy for esophageal adenocarcinoma.     

Expression of cyclooxygenase-2 in breast carcinogenesis and its relation to HER-2/neu and p53 protein expression in invasive ductal carcinoma

The purpose of this study was to evaluate cyclooxygenase-2 (COX-2) expression in the successive steps of breast carcinogenesis and to determine its correlation with HER-2/neu and p53 expression in invasive ductal carcinomas of the breast. Immunohistochemical staining with anti-COX-2 antibody was performed in normal breast tissue, usual hyperplasia, ductal carcinoma in situ, and invasive ductal carcinoma. Expression of COX-2 in invasive ductal carcinoma was correlated with immunohistochemical expression of HER-2/neu and p53 protein. COX-2 expression was found to be progressively elevated along the continuum from normal breast tissue to invasive ductal carcinoma (P<0.001). COX-2 expression significantly correlated with p53 and HER-2/neu protein expression (P<0.05 and P<0.001). On multivariate analysis, only TNM stage and elevated COX-2 expression correlated with survival. Our results suggest that COX-2 may be involved in the carcinogenesis of the breast and may be an independent prognostic indicator in patients with invasive ductal carcinoma. HER-2/neu and p53 are likely to be involved in the regulation of COX-2 expression in invasive ductal carcinomas of the breast.     

Cardiac mucosa in the remnant esophagus after esophagectomy is an acquired epithelium with Barrett's-like features

BACKGROUND: The cervical esophagus is normally lined by squamous epithelium and is usually not exposed to gastroesophageal reflux. The aims of this study were, first, to investigate whether cardiac mucosa can be acquired in the remnant cervical esophagus after esophagectomy and cervical esophagogastrostomy and, second, to characterize this mucosa if present. METHODS: The medical records of 100 patients who had undergone esophagectomy with gastric pull-up reconstruction were studied retrospectively to identify those who had biopsies from the cervical esophagus proximal to the gastroesophageal anastomosis during postoperative follow-up. The histopathology and immunohistochemical stains were reviewed to assess similarity to Barrett's mucosa (cytokeratins [CK] 7 and 20 and DAS-1), cellular proliferation (topoisomerase 2alpha), and the potential for dysplasia (cyclo-oxygenase 2 [COX-2] and ornithine decarboxylase [ODC]). RESULTS: Supra-anastomotic biopsies were performed in 20 patients. Cardiac mucosa was present in 10 of 20 (50%) patients in whom biopsies were performed. Four patients had areas of intestinal metaplasia, and dysplasia, and adenocarcinoma developed in 1 patient. The CK7/20 and DAS-1 staining of the columnar mucosa showed a pattern similar to Barrett's mucosa. Topoisomerase 2alpha protein expression was present in 50% of patients with cardiac mucosa. DAS-1 protein was expressed in cervical columnar mucosa but not in normal squamous esophagus mucosa. The cardiac mucosa stained weakly for COX-2 and ODC. CONCLUSIONS: Cardiac mucosa can be acquired. Its expression profile is similar to cardiac mucosa and intestinal metaplasia found in Barrett's esophagus, and different from normal esophageal or gastric mucosa. The development of cardiac mucosa is likely to be related to reflux of acid into the remnant cervical esophagus as the first step in the development of Barrett's esophagus. These findings are applicable to the development of similar changes at the gastroesophageal junction.     

胶质瘤间质血管增生和细胞增殖与环氧化酶-2基因的关系

目的 探讨环氧化酶-2(COX-2)基因与胶质瘤间质血管增生和细胞增殖的关系.方法 用原位杂交方法 检测胶质瘤组织中COX-2 mRNA的表达;用免疫组织化学检测同等标本中细胞增殖指数(Ki-67LI)及微血管密度(MVD)的表达;并将COX-2 mRNA与Ki-67、CD34作相关分析.结果 胶质瘤中COX-2 mRNA表达阳性率为62.69%,对照脑组织中无阳性表达.COX-2 mRNA阳性着色定位于微血管周围,呈棕黄色园环状表现.C0x-2 mRNA表达阳性率在低度恶性胶质瘤(Ⅰ、Ⅱ级)中为37.14%(14/35),而在高度恶性胶质瘤(Ⅲ、Ⅳ级)中为87.50%.在低度、高度恶性胶质瘤中MVD分别为12.54±3.11和20.31±5.49;其Ki-67LI分别为12.29±5.12和23.31±7.14.COX-2 mRNA表达为阴性和阳性胶质瘤中其MVD分别为16.90±6.48和9.46±2.15,其Ki-67LI分别为18.73±8.87和11.10±2.93.结论 COX-2 mRNA在胶质瘤组织中的表达与胶质瘤间质血管增生和细胞增殖密切相关;MVD和Ki-67U在COX-2 mRNA表达为阴性和阳性组织中比较差异有统计学意义.     

Cytokeratin 20, AN43, PGDH,and COX-2 expression in transitional and squamous cell carcinoma of the bladder

Bladder tumors from Egyptian patients with a high prevalence of bilharziasis were immunohistochemically analyzed for the expression of cytokeratin 20 (CK20), AN43, prostaglandin dehydrogenase (PGDH), and cyclooxygenase-2 (COX-2). The tumors included 26 transitional cell carcinomas (TCC), 10 squamous cell carcinomas (SCC) and 2 tumors of mixed TCC/SCC histology. Of the 28 TCC tumors, 21 (75%) expressed CK20 and 25 (89%) expressed AN43. CK20 was not expressed in any of the 10 SCC tumors and AN43 was expressed in 2 of them. PGDH was expressed in 18 (64%) of the 28 tumors with TCC histology and 1 of the 10 SCC. A subset of 21 tumors (16 TCC and 5 SCC) was tested for COX-2 expression. COX-2 was detected in 69% of the 16 TCC tumors examined but was not seen in the SCC tumors. As tumors increased in stage, the expression of these proteins changed. CK20, AN43 and PGDH decreased but COX-2 expression increased in higher stage tumors. The histologic phenotype of these cancers is reflected in their expression of these proteins and is modified further as tumors progress in stage.     

Quantitative, tissue-specific analysis of cyclooxygenase gene expression in the pathogenesis of Barrett's adenocarcinoma.

Cyclooxygenase (Cox-2) is implicated in the pathogenesis of many cancers including esophageal adenocarcinoma (EAC), whereas the role of the isoform Cox-1 in carcinogenesis is not well understood. To further elucidate the role of these factors in the development of EAC, we measured the gene expressions (mRNA levels) of Cox-2 and Cox-1 by real-time quantitative polymerase chain reaction (QRT-PCR) in tissues from normal esophagus with and without erosive gastroesophageal reflux disease (GERD), Barrett's esophagus (BE), dysplasia, adenocarcinoma, and in healthy gastric antrum. All tissues were purified by laser capture microdissection from endoscopic or surgical resection specimens. Median Cox-2 gene expression did not differ significantly among the esophageal control groups but was elevated 5-fold in BE, 8-fold in dysplasia and 16-fold in EAC compared to normal esophageal controls with no erosive GERD. Erosive GERD tissue had slightly higher median Cox-2 expression but Cox-2 expression in normal antrum was much higher than that in a normal esophagus, close to that of dysplasia. In contrast to that of Cox-2, Cox-1 expression was significantly decreased in all neoplastic tissues compared to normal controls. Cox-1 and Cox-2 expression varied over a wide range in the neoplastic tissues but over a relatively narrow range in the esophageal normal tissues. The occurrence of substantial alterations in Cox-1 and Cox-2 expression at the BE stage indicates that these are early events in the development of EAC. These results confirm the important role of Cox-2 amplification in the pathogenesis of esophageal adenocarcinoma, but the unexpected down-regulation of Cox-1 raises questions about its role in carcinogenesis.