Establishment and characterisation of six human biliary tract cancer cell lines

Human cell lines established from biliary tract cancers are rare, and only five have been reported previously. We report the characterisation of six new six biliary tract cancer cell lines (designated SNU-245, SNU-308, SNU-478, SNU-869, SNU-1079 and SNU-1196) established from primary tumour samples of Korean patients. The cell lines were isolated from two extrahepatic bile duct cancers (one adenocarcinoma of common bile duct, one hilar bile duct cancer), two adenocarcinomas of ampulla of Vater, one intrahepatic bile duct cancer (cholangiocarcinoma), and one adenocarcinoma of the gall bladder. The cell phenotypes, including the histopathology of the primary tumours and in vitro growth characteristics, were determined. We also performed molecular characterisation, including DNA fingerprinting analysis and abnormalities of K-ras, p15, p16, p53, hMLH1, hMSH2, DPC4, beta-catenin, E-cadherin, hOGG1, STK11, and TGF-betaRII genes by PCR-SSCP and sequencing analysis. In addition, we compared the genetic alterations in tumour cell lines and their corresponding tumour tissues. All lines grew as adherent cells. Population doubling times varied from 48-72 h. The culture success rate was 20% (six out of 30 attempts). All cell lines showed (i) relatively high viability; (ii) absence of mycoplasma or bacteria contamination; and (iii) genetic heterogeneity by DNA fingerprinting analysis. Among the lines, three lines had p53 mutations; and homozygous deletions in both p16 and p15 genes were found three and three lines, respectively; one line had a heterozygous missense mutation in hMLH1; E-cadherin gene was hypermethylated in two lines. Since the establishment of biliary tract cancer cell lines has been rarely reported in the literature, these newly established and well characterised biliary tract cancer cell lines would be very useful for studying the biology of biliary tract cancers, particularly those related to hypermethylation of E-cadherin gene in biliary tract cancer.     

PTEN和E-钙粘蛋白表达与胃癌浸润转移和预后的关系

目的探讨胃癌组织中PTEN和E-钙粘蛋白的表达、相互关系及意义。方法应用免疫组化SP技术检测80例胃癌组织中PTEN和E-钙粘蛋白的表达。结果胃癌组织中PTEN高表达率和E-钙粘蛋白正常表达率分别为43.8%(35/80)和41.3%(33/80),与肿瘤分化程度、浸润深度、淋巴结转移和肿瘤分期显著相关;PTEN与E-钙粘蛋白表达显著正相关,与病人预后相关,Kaplan-Meier生存曲线分析显示PTEN高表达和E-钙粘蛋白正常表达者术后累计生存率显著高于PTEN低表达和E-钙粘蛋白异常表达者,PTEN高表达和E-钙粘蛋白正常表达者术后3年、5年生存率高,Log-rank检验差异有显著性。结论胃癌组织中PTEN表达减少,E-钙粘蛋白表达异常,PTEN可能通过调控胃癌组织钙粘蛋白表达,抑制胃癌的浸润和转移,影响病人的预后。     

益肾种子丸对不明原因不孕症患者黄体中期子宫内膜E-钙黏素表达的影响

目的：通过观察益肾种子丸对不明原因不孕症患者黄体中期子宫内膜E-钙黏素表达的影响，探讨其治疗不孕症的机制。方法：采用免疫组织化学方法分别检测45例不明原因不孕症患者（研究组30例，对照组15例）及17例正常妇女（正常组）黄体中期子宫内膜E-钙黏素的表达，然后研究组连续服用益肾种子丸3个月，对照组不服药，监测排卵，指导同房，疗程结束后复查。结果：不孕症患者黄体中期子宫内膜E-钙黏素表达呈弱阳性。正常组呈强阳性，二者比较差异有显著性意义（P〈0．01）。研究组治疗后较治疗前表达增高，差异有显著性意义（P〈0．01）。结论：不明原因不孕症患者黄体中期子宫内膜E-钙黏素表达缺陷可能是导致不孕症的原因之一；益肾种子丸能增强不孕症患昔黄体中期E-钙黏素的表达，可能是其治疗不孕的机制。     

E-cadherin、β-catenin和p120ctn在所谓肺硬化性血管瘤中的表达

背景与目的所谓肺硬化性血管瘤(so-called pulmonary sclerosing hemangioma,PSH)是一种迄今未能确定其组织来源及性质的少见肺部肿瘤,多年来一直是人们研究的热点.本研究通过检测E-cadherin、β-catenin和p120ctn在PSH表面立方细胞和间质多角形细胞的免疫表型以探讨其组织来源.方法采用S P免疫组化法检测25例手术切除PSH标本及8例肺炎性假瘤标本的E-cadherin、β-catenin和p120ctn表达情况.结果25例PSH的立方细胞中,三种上皮粘附分子E-cadherin、β-catenin和p120 ctn均呈胞膜强阳性表达,β-catenin在胞膜强阳性表达的同时有胞质表达.而在多角形细胞中,E-cadherin表达缺失,β-catenin胞膜表达缺失伴部分胞质表达,p120 ctn胞质表达为主伴少量胞膜表达;三种粘附分子在多角形细胞中的表达存在异质性.血管瘤样区的腔内衬细胞E-cadherin、β-catenin和p120 ctn均呈胞质胞膜阳性表达.8例肺炎性假瘤中增生的Ⅱ型肺泡细胞E-cadherin、β-catenin和p120 ctn表达与PSH中立方细胞的表达情况相同.结论PSH中的立方细胞可能是增生的Ⅱ型肺泡细胞,而多角形细胞为肿瘤的实质细胞且缺乏分化成熟的上皮细胞所具有的E-cadherin/catenin复合体.血管瘤样区的腔内衬细胞是与立方细胞相同的上皮细胞而非血管内皮细胞.     

上皮钙粘蛋白在口腔鳞状细胞癌组织中的表达与临床预后的相关性探讨

目的：探讨上皮钙粘蛋白的表达量与口腔鳞状细胞癌（SCC）的临床病理学参数及SCC患者的预后之间的关系。方法：采用免疫组织化学染色SABC法检测43例口腔鳞状细胞癌组织标本中的蛋白表达量并分析其与临床指标的关系。结果：上皮钙粘蛋白表达与口腔鳞状细胞癌的病理学分级（P=0．024），浸润类型（P=0．009）。淋巴结转移情况（P=0．023）及患者的生存年限（P=0．0146）有着密切的关系，蛋白表达越低，病理分级越高，浸润类型越复杂，出现淋巴结转移同时患者生存时间越短。但是，上皮钙粘蛋白表达与患者的年龄，性别及肿瘤的分化度之间无统计学意义。结论：上皮钙粘蛋白是口腔鳞状细胞癌患者一种极其重要的诊断标志物和可靠的预后因子。     

Snail与E-cadherin在上皮性肿瘤中的表达及其临床意义

目的:探讨Snail与E-cadherin蛋白表达在上皮性肿瘤中的临床意义。方法:应用免疫组织化学SP法,研究150例人肝癌、乳腺癌、宫颈癌、子宫内膜癌与卵巢癌临床标本及30例相应正常组织中Snail与E-cadherin蛋白的表达及定位。结果:在上述5种组织中,Snail主要表达于癌组织细胞核、细胞质,相应正常组织往往缺如,且在肝癌、乳腺癌中Snail表达与肿瘤转移有一定的相关性,并与E-cadherin表达呈负相关。E-cadherin定位于细胞膜,在肝、乳腺的正常组织中E-cadherin强表,相应癌组织中其表达往往下降或缺如;而在宫颈癌、内膜癌与卵巢癌中,E-cadherin表达与癌变无明显相关性。结论:Snail、E-cadherin的表达在肝癌、乳腺癌中具有重要意义,Snail有可能作为肝癌、乳腺癌肿瘤转移的生物学标志。     

转移抑制因子KAI1和E-Cadherin在食管癌组织中的表达及其意义

目的 探讨KAI1和E-Cadherin在食管癌中的表达及其意义。方法 用免疫组化S-P法检测KAI1和E-cadherin蛋白在60例食管癌及对应的食管正常黏膜组织中的表达情况。结果 在食管鳞癌组织中KAI1表达（50％）显著低于食管正常黏膜组织（81．67％）（P〈0．01）。食管一常组织中E-cadherin表达（80％）显著高于癌组织（41．67％）（P〈0．01）。KAI1低表达与食管癌病理分级、淋巴结转移及食管癌浸润深度无关（P〉0．05）；E-cadherin低表达与食管癌病理分级、淋巴结转移密切相关（P〈0．05），与食管癌浸润深度无关（P〉0．05）；两者表达皆与食管癌的人体分型无关（P〉0．05）。KAI1与E-cadherin表达无明显相关性（P〉0．05）：结论 E-cadherin的低表达与食管癌的淋巴结转移有关；KAI1低表达未见对食管癌浸润转移的影响；两者无协同作用。     

上皮型钙粘附蛋白在子宫内膜癌中的表达及意义

目的　探讨上皮型钙粘附蛋白 (E -cad)在正常子宫内膜、子宫内膜不典型增生、子宫内膜癌中的表达 ,及其与临床参数间的关系。方法　应用免疫组化S -P法检测 14例正常子宫内膜、10例子宫内膜不典型增生及 5 0例子宫内膜癌中E -cad表达情况。结果　在正常子宫内膜、子宫内膜不典型增生及子宫内膜癌中 ,E -cad阳性表达率分别为 10 0 %、80 0 %、42 0 %,各组差异具有显著性 (P <0 .0 5 )。在G1、G2 、G3 子宫内膜癌中 ,E -cad阳性表达率分别为 72 2 %、30 0 %、16 7%(P <0 .0 5 )。在限于内膜组、浸润浅肌层及浸润深肌层组 ,E -cad阳性表达率分别为 6 8 4 %、35 3%、14 3%(P <0 .0 5 ) ,在临床Ⅰ、Ⅱ、Ⅲ期 ,E -cad阳性表达率分别为 6 0 9%、31 3%、18 2 %(P <0 .0 5 )。结论　E -cad异常表达与子宫内膜癌的发生发展密切相关。     

上皮型钙粘蛋白在大肠癌中的表达

目的　探讨上皮型钙粘蛋白 (E cadherin)表达与大肠癌的发生、浸润、转移的关系。方法　应用S P免疫组化方法检测 81例大肠癌组织和正常大肠粘膜组织中E cadherin的表达。结果　正常大肠癌粘膜上皮中E cadherin全部表达。大肠癌组织E cadherin的表达与癌组织分化程度、生长方式、浸润深度和淋巴结转移密切相关 (P <0 .0 1)。结论　E cadherin与肿瘤的发生、浸润、转移有关 ,可作为一种新的肿瘤标记物 ,其表达对判断预后有一定价值。     

Lobular carcinoma in situ of the breast is not caused by constitutional mutations in the E-cadherin gene

Lobular carcinoma in situ (LCIS) is an unusual histological pattern of non-invasive neoplastic disease of the breast occurring predominantly in women aged between 40 and 50 years. LCIS is frequently multicentric and bilateral, and there is evidence that it is associated with an elevated familial risk of breast cancer. Although women with LCIS suffer an increased risk of invasive breast disease, this risk is moderate suggesting that LCIS may result from mutation of a gene or genes conferring a high risk of LCIS, but a lower risk of invasive breast cancer. The high frequency of somatic mutations in E-cadherin in LCIS, coupled with recent reports that germline mutations in this gene can predispose to diffuse gastric cancer, raised the possibility that constitutional E-cadherin mutations may confer susceptibility to LCIS. In order to explore this possibility we have examined a series of 65 LCIS patients for germline E-cadherin mutations. Four polymorphisms were detected but no pathogenic mutations were identified. The results indicate that E-cadherin is unlikely to act as a susceptibility gene for LCIS.