RNA干扰技术抑制上皮性钙黏附分子表达对HO-8910细胞的影响

目的 探讨通过RNA干扰(RNAi)技术下调上皮性钙黏附分子(E-cad)表达后,对人卵巢浆液性囊腺癌HO-8910细胞的侵袭、迁移能力的影响.方法 将针对E-cad的双链小干扰RNA(siRNA)转染入HO-8910细胞中,抑制E-cad基因表达;采用免疫荧光法及Western blotting检测RNAi效果;用Transwell小室法检测RNAi后细胞侵袭、迁移能力的改变.结果 RNAi后HO-8910细胞E-cad表达显著下降,由63.7%降低为11.9%(P＜0.01),RNAi有效;细胞对细胞外基质的侵袭能力明显提高,穿过滤膜的细胞数由13.4±3.93增至42.0±4.15(P＜0.01);细胞迁移能力也明显提高,穿过滤膜的细胞数由23.24±3.71增至82.0±5.51(P＜0.01).结论 人卵巢浆液性囊腺癌的侵袭、转移等恶性行为可能与E-cad基因表达下降有关.     

β-连环蛋白与卵巢癌转移的相关性及调节机制

目的 探讨β-连环蛋白(β-catenin)与卵巢癌转移的相关性及其可能的调节机制. 方法 采用免疫荧光法、免疫印迹法及反转录-聚合酶链反应(RT-PCR)检测人卵巢浆液性囊腺癌高、低转移细胞系(HO-8910PM及HO-8910)中β-catenin基因的表达差异.为进一步探讨β-catenin调节卵巢癌转移的可能机制,将低转移卵巢癌HO-8910细胞经肝细胞生长因子(HGF)处理,应用上述方法检测β-catenin基因表达的改变;同时经Transwell小室法和划痕实验检测细胞侵袭、迁移能力的改变. 结果 β-catenin在HO-8910细胞中为高表达,在HO-8910PM细胞中为低表达,差异具有显著性(P<0.05).HGF可明显降低HO-8910细胞中β-catenin基因的表达(P<0.05)并促进细胞侵袭(P<0.05)和迁移. 结论 卵巢癌细胞的侵袭、迁移等恶性行为可能与β-catenin基因表达下降有关.经由HGF/c-Met的信号很可能是β-catenin介导的黏附功能的重要调节者.     

Caveolin-1与人卵巢癌转移的相关性及其机制的研究

目的探讨窖蛋白-1(caveolin-1)与卵巢癌转移的相关性及其可能的调节机制。方法采用免疫荧光法、蛋白印迹实验及逆转录聚合酶链反应检测人卵巢浆液性囊腺癌高低转移细胞系(HO-8910PM及HO-8910)中caveolin-1基因的表达差异。表皮生长因子(EGF)处理低转移卵巢癌HO-8910细胞后,经上述方法检测caveolin-1基因表达的改变。同时经Transwell小室法检测细胞侵袭、迁移能力的改变。结果 caveolin-1在HO-8910细胞中为高表达,在HO-8910PM细胞中为低表达(P0.01)。EGF可明显降低HO-8910细胞中caveolin-1基因的表达(P0.01)并促进细胞侵袭(P0.01)和迁移(P0.05)。结论卵巢癌细胞的侵袭、迁移可能与caveolin-1基因表达下降有关,经由EGF/EGFR的信号很可能是caveolin-1介导的卵巢癌转移抑制的重要调节者。     

卵巢浆液性、黏液性囊腺癌DNA拓扑异构酶的表达

目的：探讨DNA拓扑异构酶在卵巢浆液性,黏液性囊腺癌中的表达及与细胞增殖,凋亡的关系。方法：应用免疫组化S－P法检测石蜡包埋的71例卵巢浆液性,黏液性囊腺癌组织切片中DNA拓扑异构酶Ⅱ（DNA topoisomeraseⅡ,TOPOⅡ）表达和细胞增殖指数（Ki-67 labeling index,Ki-67 LI)和应用原位末端标记技术（TUNEL）检测细胞凋亡指数（apoptosis index,AI).结果：TOPOⅡ高表达为36．6％（26／71）,与组织学类型,FIGO分期和组织学分级差异无显著性（P＞0．05）;Ki-67 LI高表达为43．7％（31／71）,与FIGO分期差异有显著性（P＜0．001）;AI高表达为25．6％（18／71）,与组织学分级,FIGO分期和组织学分级均差异无显著性（P＞0．05）,但AI与TOPOⅡ表达有相关（P＜0．01）。结论：应用免疫组化方法检测组织中TOPOⅡ和Ki-67LI水平,对卵巢浆液性,黏液性囊腺癌化疗方案选择和预后判断具有实用意义。     

浆液性和黏液性卵巢囊腺癌患者线粒体结构和紧密连接蛋白表达的差异比较

目的:探讨浆液性卵巢囊腺癌(SC)与黏液性卵巢囊腺癌(MC)细胞线粒体结构及紧密连接(TJ)蛋白occludin/ZO-1表达异常的差异及可能机制。方法:临床确诊30例卵巢囊腺癌患者中15例为SC(SC组),15例为MC(MC组),另选3例子宫肌瘤患者进行对照(对照组)。采用电镜观察各组线粒体超微结构;采用体视学方法分析线粒体面数密度(Nm)、体密度(Vvm)和外膜面密度(Sv);采用免疫组化方法检测各组卵巢上皮细胞occludin/ZO-1蛋白的阳性单位(PU)及其强弱程度(用积分光密度值IOD表示)。结果:电镜下,对照组线粒体及内质网均未见异常;MC组线粒体体积小、数量较多,线粒体Nm、Vvm和Sv较对照组明显降低(P0.05);SC组线粒体体积大,脊断裂显著,线粒体Nm、Vvm和Sv较MC组显著升高(P0.01)。免疫组化分析显示MC组occludin和ZO-1PU明显高于对照组(24.23±4.74Vs 21.07±5.51、37.07±3.58Vs 20.46±4.89,P均0.05),IOD明显低于对照组(72.39±9.86Vs 92.38±7.17、73.51±6.86Vs 80.34±7.04,P均0.05);SC组occludin和ZO-1PU明显低于对照组(11.50±4.15Vs 21.07±5.51、16.24±3.61Vs 20.46±4.89,P均0.05),IOD明显高于对照组(101.38±6.16Vs 92.38±7.1、92.78±3.07Vs 80.34±7.04,P均0.01)。结论:SC及MC两种卵巢囊腺癌患者线粒体结构和体视学特征明显不同,可能与TJ蛋白的occludin/ZO-1表达异常有关。     

E-cadherin的表达与肝细胞癌侵袭和转移的关系

目的研究上皮性钙黏蛋白（E-cadherin）在人肝细胞癌（HCC）中的表达情况,探讨其与肝癌的相关性。方法选择56例有完整随访资料的肝细胞癌及相应癌旁组织标本、20例正常肝组织标本,用RT-PCR方法检测E-cadherinmRNA的表达;用免疫组织化学方法检测E-cadherin的表达。结果①E-cadherin在肝细胞癌组织中的表达显著低于癌旁组织和正常组织（P〈0．05）,而在癌旁组织和正常组织中的表达无差异;②E-cadherin在肝癌组织中的表达与术后复发时间呈正相关（P〈0．05）,与病理分期呈负相关（P〈0．05）;③癌组织中E-cadherin表达与肝外转移呈负相关（P〈0．05）;④癌旁组织中E-cadherin表达与术后复发时间呈正相关（P〈0．05）。结论E-cadherin表达缺失或下调与肝癌的分化程度、侵袭转移能力和复发倾向相关,对肝癌临床转归的评估有一定指导意义。     

卵巢浆液性肿瘤中IQGAP1与E-cadherin表达的检测

目的探讨上皮性钙黏素(E-cadherin)与具有IQ结构域的人Ras GTP激活蛋白相关蛋白1(IQGAP1)在卵巢浆液性肿瘤中的表达及其临床意义。方法分别用免疫组织化学SP法和Western blot法检测20例卵巢浆液性囊腺癌、10例卵巢交界性浆液性囊腺瘤、10例卵巢浆液性囊腺瘤以及10例正常卵巢组织中E-cadherin和IQGAP1的蛋白表达。结果 E-cadherin蛋白在浆液性囊腺瘤中表达最高,明显高于正常卵巢组织和浆液性囊腺癌组织(P0.05),其均值高于交界性囊腺癌,无统计学意义。IQGAP1在卵巢浆液性囊腺癌中表达较正常卵巢组织、良性及交界性肿瘤组织均增高(P0.05);免疫组织化学染色证实IQGAP1在浆液性囊腺癌中以胞膜表达为主,而在良性肿瘤中以胞质表达为主。结论 E-cadherin和IQGAP1在卵巢浆液性瘤高表达,可联合用于卵巢浆液性肿瘤的免疫组化诊断。     

miRNA-200c在IIIc期卵巢浆液性囊腺癌中的表达及其参与癌转移中的作用

研究miR-200c在IIIc期卵巢浆液性囊腺癌中的表达格局,探索其参与转移癌形成的相关机制。选取2010年1月至2013年12月上海交通大学医学院附属仁济医院妇科卵巢肿瘤组织标本(IIIc期卵巢浆液性囊腺癌40例(其中选择自身配对的原发灶40例、转移灶40例)、卵巢浆液性囊腺瘤30例为对照),采用real time-qPCR法检测miR-200c的表达;采用CCK8法、Transwell小室分别检测细胞过表达miR-200c后增殖、迁移和侵袭能力的改变;采用western blot法检测相关蛋白表达量。结果发现:(1)IIIc期卵巢浆液性囊腺癌转移灶肿瘤与原发灶肿瘤相比,miR-200c表达是升高的(490.37±78.32cf.157.46±17.21),P0.01;而且这两者均高于对照组(卵巢浆液性囊腺瘤)的表达(25.32±10.17),P0.01。SKOV3细胞过表达miRNA-200c后(2)细胞迁移能力降低(35.6±4.3cf.18.6±1.9),P0.05;侵袭能力降低(26.7±3.1cf.15.4±2.4)P0.05;但是增殖能力不改变。(3)在蛋白水平检测ZEB1蛋白表达下降(0.3023±0.0251cf.0.6753±0.0262)P0.05、E-cad蛋白表达升高(1.274±0.0331cf.0.6140±0.0460)P0.05、Vimentin蛋白表达降低(0.0957±0.0049cf.0.1767±0.0158)P0.05。可见,miR-200c在IIIc期卵巢浆液性囊腺癌组织样本中表达增高,检测miR-200c的表达格局对于临床上监测上皮性卵巢癌的临床进展和转移有重要意义;也为开拓过表达miR-200c作为卵巢癌治疗手段,提供了理论与实验依据。     

基于生物信息学分析PI3在卵巢浆液性囊腺癌中的表达及临床意义

目的 探究PI3基因在卵巢浆液性囊腺癌（OV）中的表达及临床意义。方法 利用多种生物信息学分析在线工具,基于GEO、TCGA、GTEx、GEA数据库挖掘PI3基因在OV的表达、生物学作用及临床意义。结果 PI3在OV组织的表达量高于正常卵巢组织（P<0.05）;OV患者中,PI3低表达组总生存期和无复发生存期比高表达组长（P<0.001）;在免疫细胞浸润方面,中性粒细胞水平与PI3在OV中的表达呈正相关,Th1细胞、Th2细胞、B细胞、嗜酸性粒细胞水平对OV患者生存有影响且与PI3表达高低有关;PI3参与了炎症反应、白细胞活化等生物学过程,参与了PI3K-Akt、Toll样受体等信号通路。结论 PI3基因在OV中高表达,可作为OV患者诊断、治疗的潜在靶点。     

基于癌症基因组图谱筛查卵巢浆液性囊腺癌相关基因

目的:利用全基因组表达谱芯片筛查与卵巢浆液性囊腺癌发生相关的基因,对在卵巢浆液性囊腺癌发生过程中可能参与的基因间的信号转导通路进行分析。方法:选取癌症基因组图谱(TCGA)数据库中卵巢浆液性囊腺癌的Affymetrix Gene Chip Human Exon 1.0 ST Array数据共16张,分别为卵巢浆液性囊腺癌组8张和正常组8张,筛选出差异表达基因,并进行基因本体(gene ontology,GO)分析和信号通路分析,构建卵巢浆液性囊腺癌相关基因间的信号转导通路,分析网络中具有重要作用的基因。结果:共筛选出1 144个在卵巢癌中差异表达的基因,其中表达上调的基因有747个,表达下调的基因有397个。GO分析得到上调差异基因的显著性功能分析结果362项,下调差异基因的显著性功能分析结果 160项(P0.05)。其中包括与肿瘤发生相关的基因功能有细胞周期、DNA复制、细胞增殖、细胞凋亡、细胞黏附等。信号通路分析得到45个显著上调信号通路和14个显著下调信号通路(P0.05)。其中参与肿瘤发生相关的信号通路主要有细胞周期、P53信号通路、DNA复制、肿瘤中的信号通路、PI3K-Akt信号通路、ECM-receptor信号通路、细胞黏附因子、细胞凋亡等。挑选显著性基因功能和信号通路分析的交集基因229个,构建显著性GO与信号通路基因间信号转导网络。分析发现CDK1、PLK1、MCM3和PGK1这4个基因在卵巢癌的基因调控网络中具有重要作用。结论:卵巢浆液性囊腺癌中有大量差异表达基因,差异表达的基因在多个与肿瘤发生密切相关的信号通路中发挥重要的调控作用。     

妊娠滋养细胞肿瘤中E-cadherin和增殖细胞核抗原的表达

目的：研究妊娠滋养细胞肿瘤(GTT)中E-cadherin与PCNA的表达意义。方法：采用免疫组化SP法检测了妊娠滋养细胞肿瘤中E-cadherin与PCNA的表达。结果：E-cadherin在正常早期绒毛(NP)及其肿瘤(CM、IM、OCA)中表达率分别为56．29％、42．07％、19．30％、7．14％,各组间差异非常显著。PCNA在NP、CM、IM和OCA中表达率分别为10．40％、20．76％、53．60％、51．95％,各组间差异非常显著。E-cadherin与PCNA阳性率呈负相关。结论：E-cadherin和PCNA的检测有助于良恶性GTT的鉴别;E-cadherin表达减弱或消失促进了GTT的增殖活性。     

Expression of E-cadherin and vimentin in oral squamous cell carcinoma

The aim of the study is to determine the levels of E-cadherin, vimentin expression in tumor tissues from patients with oral squamous cell carcinoma (OSCC), and the relationship between the expression of E-cadherin, vimentin and epithelial-mesenchymal transition, in order to explore its values for predicting the invasion and metastasis of oral squamous cell carcinoma, short survival of patients in many types of cancer. E-cadherin and vimentin expression of 10 benign and 42 OSCC tumor tissues was examined by immunohistochemical staining. E-cadherin is positively expressed in normal oral mucosa epithelium, but vimentin expression is not found in normal oral mucosa epithelia; the E-cadherin and vimentin were expressed in 26 of 42 (61.9%) and 16 of 42 (38.1%), respectively. No statistically difference was found for E-cadherin and vimentin expression in patients with different age, gender and tumor location, E-cadherin and vimentin expression was significantly associated with lymph node metastasis and tissue location (P < 0.05); E-cadherin expression was also significantly associated with tumor stage (P < 0.05); there are significantly difference between infiltrative margin and central area in patients with oral squamous cell carcinoma for E-cadherin and vimentin positive expression (P < 0.05). E-cadherin and vimentin positive expression was associated with tumor metastasis of oral squamous cell carcinoma. Our study preliminarily confirmed that EMT phenomenon is existed during the development of oral squamous cell carcinoma. Co-evaluation of E-cadherin and vimentin might be a valuable tool for predicting OSCC patient outcome.     

Expression of E-cadherin (E-CD) as related to other prognostic factors and survival in breast cancer

A series of 208 breast cancer biopsies were analysed immunohistochemically for expression of E-cadherin (E-CD). Altogether, 72 per cent of the tumours showed E-CD positivity in over 50 per cent of cells, the staining being heterogeneous in nearly all tumours. In only 16 per cent of ductal carcinomas was positive staining seen in less than 1 per cent of cells. Expression of E-CD was not related to tumour diameter, nodal status, metastasis at diagnosis, histological grade, DNA ploidy, S-phase fraction, nuclear area, mitotic frequency, or PR content. There was a significant relationship between expression of E-CD, histological type (P=0.01), the proportion of intraductal growth (P=0.008), the density of tumour-infiltrating lymphocytes (P=0.0007), ER content (P=0.012), and morphometric nuclear factors (P=0.02). Expression of E-CD showed a weak association with a high survival probability (P=0.02), while the relation to recurrence-free survival was not significant (P=0.11). In axillary lymph node-negative tumours, E-CD expression was not related significantly to survival (P=0.11) or to recurrence-free survival (P=0.06). In multivariate analysis, E-CD expression had no independent prognostic value, while the axillary lymph node status, tumour diameter, patient age, and mitotic frequency were independent prognostic factors. The results indicate that E-CD expression is related to several histological features in breast cancer, but has no independent prognostic value over standard prognostic factors.     

P120-catenin和E-cadherin的协同表达与肺癌的恶性程度相关

目的方法结果结论探讨p120-catenin(p120ctn)和E-cadherin(E-cad)在非小细胞肺癌中的表达是否具有相关性。方法免疫组织化学S-P法、RT-PCR及WesternBlot等检测肺癌组织中p120ctn和E-cad的表达情况。结果p120ctn和E-cad在肺癌组织中的表达明显低于正常肺组织,并且p120ctn的异常表达与E-cad的异常表达呈正相关。肺癌组织中p120ctn和E-cad的异常表达与肺癌的高分期,低分化和淋巴结转移明显相关。与正常支气管上皮细胞HBE相比,同源的BE1和LH7细胞中都显示出p120ctn、E-cad的表达下降。但在转移能力较高的BE1细胞系中这种异常表达的现象尤为明显。结论肺癌中p120ctn和E-cad的低表达可能与肺癌的恶性度有关。     

Expression of catenins and E-cadherin during epithelial restitution in inflammatory bowel disease

Catenins are cytoplasmic proteins associated with E-cadherin, the prime mediator of cell–cell adhesion. Perturbation in any of these molecules results in altered intercellular adhesion, cell differentiation, and increased migration. In this study, the expression and cellular localization of catenins and E-cadherin in inflammatory bowel disease were examined. The expression of E-cadherin; α-, β-, and γ-catenin; and p120 was evaluated immunohistochemically in 31 paraffin-embedded colonic specimens from 21 patients with ulcerative colitis and Crohn's disease. Loss of normal membranous E-cadherin and α-catenin staining was detected at the mucosal edges around epithelial ulcerations in all cases of active ulcerative colitis and in 50 per cent of cases with active Crohn's disease. Reduced expression of p120 protein was also found at the margins of ulcerated mucosa in all cases of active ulcerative colitis and in 75 per cent of those with active Crohn's disease. There was a statistically significant correlation between the expression of E-cadherin, α-catenin and p120 and disease activity. There were no changes in β- and γ-catenin expression in either ulcerative colitis on Crohn's disease. These findings indicate that altered expression of E-cadherin, α-catenin, and p120 occurs during mucosal ulceration in inflammatory bowel disease. These changes may be involved in promoting cell migration during epithelial restitution of the gastrointestinal mucosa. © 1998 John Wiley & Sons, Ltd.