E钙黏着素与波形蛋白在膀胱癌中的表达及意义

目的检测E钙黏着素（E—cadherin）和波形蛋白（Vimentin）在膀胱癌组织中表达及意义：方法采用免疫组织化学技术检测60例膀胱移行细胞癌组织中E—cadherin和Vimentin的表达,分析二者的相关性及其与膀胱癌病理分级的关系。结果E—cadherin和Vimentin在不同病理分级的膀胱癌组织中的表达差异有统计学意义（P〈o．01）,E-cadherin异常表达与Vimentin阳性表达成正相关（r=0．330,P〈20．05）。结论初步证实在膀胱癌的发生、发展中可能存在上皮间质转化（EMT）现象;联合检测E-cadherin及Vimentin对判断膀胱癌患者的预后具有一定的临床价值。     

Caveolin-1和E-cadherin在肾细表达及其意义胞癌中的

目的：探讨Caveolin-1、E—cadherin在肾细胞癌中的表达及其临床意义。方法：采用免疫组织化学EnVision法检测41例肾细胞癌切除标本中Caveolin-1和E—cadherin的表达，并分析表达情况与肿瘤的生物学行为的关系。结果：Caveolin-1在肾细胞癌中的表达阳性率为41．5％，与肿瘤分期、Fuhrman分级及患者年龄呈正相关（P〈0．05）；E-cadherin表达阳性率为24．4％，低于癌旁肾组织（50％），E—cadherin表达与肿瘤细胞核分级相关（P〈0.05），与其他临床病理参数之间无相关性。Caveolin-1和E-cadherin两者之间表达无相关性。结论：Caveolin-1过表达及E-cadherin的异常表达，可能在肾细胞癌的生长、分化中发挥重要作用。     

E-cadherin反义寡核苷酸对肿瘤细胞浸润能力影响的研究

目的 了解上皮型钙黏附分子（E—cadherin）下调对肿瘤细胞浸润能力的影响。方法 选用人胰腺癌细胞株JHP-1，经E—cadherin反义寡核苷酸（ASODN）作用，应用免疫细胞化学染色、Westernblot法检测细胞E—cadhefin蛋白表达情况，细胞浸润MTT比色法检测该细胞对基底膜的浸润能力。结果 经E—cadherinASODN作用后，JHP-1细胞膜E—cadherin表达减弱，E—cadherin蛋白含量与对照组相比明显降低；JHP-1细胞对基底膜浸润能力明显高于对照组（P〈0．001）。结论 E—cadherin与肿瘤细胞的浸润能力有关。     

黏附分子T-cadherin表达对胶质母细胞瘤C6细胞的生抑制作用

目的 探讨T—cadherin分子表达对胶质母细胞瘤C6细胞增殖的影响。方法 利用脂质体将pcDNA3和pcDNA3．T—cadherin表达质粒转染C6细胞，以克隆形成试验和细胞增殖试验研究T-cadherin表达对C6细咆增殖的影响。结果 转pcDNA3．T-cadherin质粒的C6细胞形成的细胞克隆数显著少于转染pcDNA3载体质粒的C6细胞形成的克隆数，两者差异有统计学意义（P〈0．01）。转染后表达T—cadherin分子C6细胞的生长速度明显慢于转染空载体不表达T—cadherin的C6克隆（P〈0．01），且T—cadherin表达水平与C6细胞增殖速度呈负要关。结论 T—cadherin分子表达显著抑制胶质母细胞瘤C6细胞的增殖。     

E-钙黏蛋白和nm23-H1在肝细胞性肝癌中的表达及其临床意义

目的探讨E-钙黏蛋白（E—cadherin）和nm23-H1在肝细胞性肝癌（HCC）中的表达及其与肝癌浸润转移之间的关系。方法应用免疫组化S—P技术，检测50例肝癌组织中E—cadherin和nm23-H1的表达，分析与肝癌侵袭转移之间的关系。结果50例肝癌组织中E—cadherin和nm23-H1的表达与HCC的Edmondson分级、侵袭转移能力密切相关，而与患者性别、年龄、肿瘤大小无关。E-cadherin和nm23-H1的表达具有高度协同性。结论E—cadherin和nm23-H1的表达能反映HCC的侵袭转移能力。联合检测两种蛋白在肿瘤组织中的表达，可以更准确判断HCC的恶性程度和生物学行为。     

保罗样激酶1与上皮钙黏蛋白在肝癌中的表达及预后价值

目的研究比较肝细胞肝癌中保罗样激酶1（PLKl）及上皮钙黏蛋白（E-cadherin）的表达与临床病理因素关系及对预测肝癌移植术后复发的价值。方法应用RT．PCR检测PLKl及E-cadherin在转录水平的表达趋势;应用免疫组织化学检测两者蛋白水平的表达情况,并与临床病理因素及术后无瘤生存期、复发率进行比较分析。结果在转录水平方面,50对肝癌及癌旁冰冻标本中PLKl及E—cadherin的表达阳性率分别为90．0％与96．0％,PLKlmRNA癌高于癌旁,E—cadherin则无趋势;50例肝癌石蜡标本中PLKl阳性表达及E—cadherin低表达的阳性率分别为60．0％与50．0％,PLKl阳性表达仅与术前血清AFP相关,E—cadherin降低与临床病理因素均无相关性,PLKl阳性表达与E—cadherin低表达之间相关,Kaplan—Meier分析结果示PLKl、E—cadherin、肝癌大小、门静脉癌栓、Edmondson分级、术前血清AFP等均与复发相关,Cox回归分析示仅PLKl有统计学意义。结论PLKl阳性表达与E—cadherin低表达预示肝癌移植术后有较高的复发率,且PLKl可作为复发的独立风险因素。     

科萨奇-腺病毒受体抑制肿瘤细胞侵袭迁移

科萨奇-腺病毒受体(coxsackieandadenovirusreceptor, CAR)在肿瘤腺病毒载体基因治疗过程中起着关键性的作用。 近年来新的研究发现,在前列腺癌和膀胱癌进展过程中存在 着CAR表达的丢失。在胚胎形成和组织分化过程中,CAR 的表达也不尽相同。AnsgarBr櫣ning等研究发现CAR的表达在人类肿瘤细胞黏附及运动过程中起着重要的作用。研究 者在E cadherin表达缺失的卵巢癌细胞系A2780和宫颈癌细 胞系CaSki中成功建立了CAR稳定表达的细胞亚克隆。当 不表达CAR的A2780母细胞再次表达CAR时,导致细胞- 细胞接触形成和细胞聚集小簇的出现。而…     

STAT3及其磷酸化STAT3和E钙黏蛋白在结直肠癌组织中的表达及意义

目的探讨结直肠癌组织中STAT3及其磷酸化STAT3（P-STAT3）和E钙黏蛋白（E—cadherin）表达的相关性及其与肿瘤浸润转移的关系。方法用免疫组织化学Elivision^TM plus法检测江西赣南医学院第一附属医院病理科存档的50例结直肠癌组织及相应癌旁组织中STAT3、P-STAT3及上皮细胞间质化（EMT）相关蛋白E—cadherin的表达,并分析其与结直肠癌临床病理特征的关系。结果50例结直肠癌组织中STAT3和p-STAT3及E—cadherin的阳性表达率分别为72％（36／50）、76％（38／50）和26％（13／50）,相应癌旁组织中的表达则分别为24％（12／50）和26％（13／50）及68％（34／50）,结直肠癌组织STAT3和p-STAT3的表达明显高于相应癌旁组织,而E—cadherin的表达则明显低于癌旁组织（均P〈0．05）。STAT3、P-STAT3及E．cadherin表达与肿瘤浸润深度、分化程度、肿瘤大小、淋巴结转移及TNM分期有关（均P〈0．05）。STAT3和P—STAT3蛋白在结直肠癌中的表达与E-cadherin呈显著负相关（均P〈0．05）。结论STAT3和p-STAT3可能通过对E—cadherin的抑制作用进而引起EMT现象,从而导致结直肠肿瘤的发生和进展。     

c-met、E-钙黏蛋白、β-连环蛋白在肝细胞癌中的表达及意义

目的 探讨c-met、E-钙黏蛋白（E—cadherin）、β-连环蛋白（β-catenin）在肝细胞癌（HCC）侵袭转移中的作用,旨在初步明确其在HCC预后判断中的作用。方法 采用免疫组化（ABC—HRP）对47例肝细胞癌手术切除标本中c-met、E-cadherin、β-catenin的表达进行了检测,评估c—met的表达与HCC临床病理因素的关系及与E-cadherin、β-catenin表达的关系。结果 c—met、E—cadherin、β-catenin在肝细胞癌组织中阳性表达率分别为38．3％、59．6％、38．3％,在正常肝组织中分别为12％、84％、64％,两者比较有显著性差异（P〈0．05）。c—met阳性表达率在肝细胞癌的转移（P〈0．05）、淋巴浸润（P〈0．05）、包膜形成（P〈0．05）以及细胞分化方面有显著性差异（P〈0．05）;而在性别、年龄、肿瘤的大小、分期方面则没有差异。E—cadherin、β-caienin的表达在肝细胞癌的转移（分别为P=0．032,P=0．022）、包膜形成（分别为P=0．016,P=0．034）方面的有显著性差异;c—met表达与E—cadherin、β-catenin的异常表达比较有显著性差异（分别为P=0．023,P=0．016）。结论 上述结果提示在HCC的转移中,c—met可能是通过HGF／c—met通路影响E-cadherin、β-catenin的改变来促进细胞的扩散进而出现转移,对于HCC的转移、复发及预后判断具有意义。     

终末期糖基化终产物通过转化生长因子依赖和非依赖途径介导NRK52E细胞转分化和胶原Ⅰ的合成

目的探讨终末期糖基化终产物（AGEs）介导肾小管上皮细胞转分化和胶原（Col）Ⅰ合成的分子机制。方法体外培养正常大鼠近端肾小管上皮细胞系（NRK52E），应用自制的AGE-牛血清白蛋白（BSA）刺激NRK52E细胞。免疫细胞化学方法检测不同时间磷酸化（P）Smad2／3核转位情况。ELISA方法检测细胞培养上清TGF-β1的水平。RT-PCR方法检测α-平滑肌肌动蛋白（SMA）、E-钙黏着糖蛋白（cadherin）和ColⅠmRNA表达。Western印迹检测α-SMA、E-cadherin和ColⅠ蛋白的表达。同时观察TGF-β1中和抗体对AGE-BSA上述效应的阻断作用。结果基础状态下，NRK52E细胞存在低水平p-Smad2／3核表达（16％）。与BSA对照组比较，AGE—BSA以时间依赖方式上调NRK52E细胞p-Smad2／3核转位，其高峰出现在30min（68％比30.5％，P〈0．01）和24h（76％比31．3％，P〈0．01）。AGE—BSA显著上调α-SMA和ColⅠmRNA和蛋白表达；下调E-cadherin mRNA和蛋白的表达；并能促进NRK52E细胞合成和分泌TGF-β1。TGF-β1中和抗体能明显抑制AGE—BSA介导的24hp-Smad2／3核转位（25．2％，P〈0．01），但不能阻抑30min活化高峰；能明显抑制AGE-BSA介导的α-SMA和ColⅠmRNA和蛋白表达．以及显著地上调E-cadherin mRNA和蛋白的表达。结论AGEs通过TGF-β依赖和非依赖途径诱导肾小管上皮细胞Smads信号通路活化，促进其向肌成纤维母细胞转分化和ColⅠ的合成。     

LIN28A和LAMP1在膀胱癌细胞系中的表达及意义

目的:探讨LIN28A和LAMP1在膀胱癌细胞系中表达情况,以及两者之间的关系,推测其可能临床意义及对肿瘤进展的影响。方法:采用RT-PCR检测膀胱癌细胞系LIN28A、LIN28B和LAMP1表达,免疫荧光检测LIN28A和LAMP1二者蛋白表达定位;LIN28A敲减后通过qRT-PCR检测LAMP1的mRNA表达变化。结果:5个癌细胞系T24、UM-UC3、J82、5637和SW780和正常移行上皮细胞系SV-HUC-1均表达LIN28A,其中J82也表达LIN28B;5个癌细胞系均表达LAMP1,SV-HUC-1不表达LAMP1;LIN28A和LAMP1蛋白均定位在胞浆;LIN28A敲减后对LAMP1的mRNA表达变化无明显影响,相应蛋白变化需要进一步验证。结论:4个膀胱癌细胞系T24、5637、UM-UC3和SW780可以用于LIN28A与肿瘤相关的机制研究,而J82可用于LIN28B的机制研究。LIN28A对肿瘤细胞和干细胞的调控方面可能具有相似性,敲减后对其靶点mRNA表达量无明显影响,LAMP1蛋白可能对肿瘤细胞侵袭转移具有抑制作用。     

膀胱癌细胞RT4和253J中转移相关蛋白的差异表达及意义

目的 检测转移相关蛋白上皮型钙黏素(E-cadherin)和波形蛋白(Vimentin)在膀胱癌细胞RT4和253J中的表达,从细胞黏附性和骨架蛋白角度探讨膀胱癌恶性进展的分子机制.方法 用Western blot法测定RT4和253J两种细胞中E-cadherin和Vimentin的表达差异,Millicell小室检测RT4和253J的体外侵袭能力,分析RT4和253J的细胞特性.结果 E-cadherin在RT4细胞中高表达、在253J细胞中低表达,而Vimentin在RT4细胞中表达缺失、在253J细胞中表达很高;Millicell证实253J细胞穿膜数较RT4细胞显著增多.结论 人膀胱癌细胞RT4和253J的E-cadherin、Vimentin表达及两种细胞的侵袭潜能差异无统计学意义,提示这两种蛋白的表达差异在促进或抑制膀胱癌的恶性进展中起重要作用.

Abstract：

Objective To detect the expression of metastasis-associated proteins in bladder cancer cells RT4 and 253J and explore the molecular mechanisms of malignant transition of bladder cancer. Methods The expression of E-cadherin and Vimentin in RT4 and 253J cells was detected by Western blotting.Millicell polycarbonate filter was used to analyze the invasive potency of RT4 and 253J cells. Results E-cadherin was detected with high expression in RT4 cells, whereas a very low expression in 253J cells:Vimentin was not detected in RT4 cells, but a high expression in 253J cells. Conclusion There was remarkable difference in the expressions of E-cadherin and Vimentin in RT4 and 253J cells, and there was different invasion characteristic in these two cell liness. E-cadherin and Vimentin play a key role in the malignant transition of bladder tumor.     

miR-451对膀胱癌细胞迁移、侵袭及E-cadherin和Vimentin基因表达的调控作用

目的研究miR-451对膀胱癌细胞迁移、侵袭及E-钙黏蛋白(E-cadherin)和波形蛋白(Vimentin)基因表达的调控作用。方法培养人膀胱癌细胞株T24、5637、SW790并采用荧光定量PCR检测miR-451的表达量;T24细胞随机分为不转染模拟物的对照组、转染NC模拟物的NC组、转染miR-451模拟物的miR-451组,采用荧光定量PCR检测miR-451的相对表达量,采用Western blot检测E-cadherin和Vimentin的相对表达量,采用Transwell检测细胞的迁移、侵袭能力。结果T24细胞中miR-451的相对表达量均低于5637细胞、SW790细胞;在T24细胞中,miR-451组的miR-451相对表达量、E-cadherin相对表达量明显高于对照组、NC组,迁移能力、侵袭能力均明显弱于对照组、NC组,Vimentin相对表达量明显低于对照组、NC组。结论miR-451能够抑制膀胱癌细胞的迁移、侵袭且该抑制作用与增加E-cadherin表达、减少Vimentin表达有关。     

MTUS2-AS2在膀胱癌组织中的表达及对膀胱癌细胞增殖和侵袭的影响

目的:探讨长链非编码RNA(lncRNA)MTUS2-AS2在膀胱癌组织中的表达及其对膀胱癌细胞增殖和侵袭的影响。方法:荧光实时定量PCR(qRT-PCR)检测63例膀胱癌组织和癌旁组织、膀胱癌细胞系(BIU-87、J82、T24、5637)和正常膀胱上皮细胞系SV-HUC-1中的MTUS2-AS2表达。以MTUS2-...     

Estrogen receptor β (ERβ) is a novel prognostic marker of recurrence survival in non-muscle-invasive bladder cancer potentially by inhibiting cadherin switch

Objective

The function and significance of estrogen receptor β (ERβ) in bladder cancer remains a field of hot debate. In this study, we aimed to (a) evaluate ERβ as a novel prognostic marker of recurrence free survival; and (b) digest the underlying mechanism by elucidating the relationship between ERβ expression and cadherin switch.

Methods

We examined the expression levels of ERβ, E-cadherin and N-cadherin in 42 initial non-muscle-invasive urothelial bladder carcinomas via immunohistochemistry. Correlation analysis was performed among ERβ expression, cadherin switch and recurrence free survival. Moreover, in vitro studies were performed to validate the identified correlation using two bladder cancer cell lines RT4 and 253J. Upon stimulation with an ERβ selective agonist diarylpropionitrile, E-cadherin, N-cadherin expressions; cell migration and invasion capacity were assessed.

Results

Expression of ERβ protein was seen in 34 bladder cancer cases (80.9 %), and 21 (50 %) specimens showed non-cadherin switch (positive E-cadherin and negative N-cadherin). ERβ expression and the non-cadherin switch are both accompanied with better recurrence free survival. Also, the least ERβ expression was observed in specimens that undergo cadherin switch. Moreover, these results were consistent with our observations in bladder cancer RT4 and 253J cell lines studies. Diarylpropionitrile stimulation resulted in an increase in E-cadherin, a decrease in N-cadherin expression and abolished cell migration and invasion.

Conclusion

ERβ is a prognostic marker of recurrence free rate in non-muscle-invasive bladder cancer, potentially through suppressing cadherin switch, and may act as a potential target for bladder cancer therapy.     

VEGF在膀胱癌细胞中的表达

探讨血管内皮生长因子在膀胱肿瘤发生发展中的作用。方法采用逆转录聚合酶链反应方法检测三种人膀胱癌细胞系和正常膀胱组织中ＶＥＧＦｍＲＮＡ的表达,同时用免疫化学方法检测三种人膀胱癌细胞系ＶＥＧＦ蛋白表达。结果三种膀胱纱均有ＶＥＧＦｍＲＮＡ的表达,正常膀组织无表达;     

Estrogen receptor β (ERβ) is a novel prognostic marker of recurrence survival in non-muscle-invasive bladder cancer potentially by inhibiting cadherin switch

Objective

The function and significance of estrogen receptor β (ERβ) in bladder cancer remains a field of hot debate. In this study, we aimed to (a) evaluate ERβ as a novel prognostic marker of recurrence-free survival and (b) digest the underlying mechanism by elucidating the relationship between ERβ expression and cadherin switch.

Methods

We examined the expression levels of ERβ, E-cadherin and N-cadherin in 42 initial non-muscle-invasive urothelial bladder carcinomas via immunohistochemistry. Correlation analysis was performed among ERβ expression, cadherin switch, and recurrence-free survival. Moreover, in vitro studies were performed to validate the identified correlation using two bladder cancer cell lines RT4 and 253 J. Upon stimulation with an ERβ-selective agonist diarylpropionitrile, E-cadherin, N-cadherin expressions; cell migration, and invasion capacity were assessed.

Results

Expression of ERβ protein was seen in 34 bladder cancer cases (80.9%), and 21 (50%) specimens showed non-cadherin switch (positive E-cadherin and negative N-cadherin). ERβ expression and the non-cadherin switch are both accompanied with better recurrence-free survival. Also, the least ERβ expression was observed in specimens that undergo cadherin switch. Moreover, these results were consistent with our observations in bladder cancer RT4 and 253 J cell lines studies. Diarylpropionitrile stimulation resulted in an increase in E-cadherin, a decrease in N-cadherin expressions and abolished cell migration and invasion.

Conclusion

ERβ is a prognostic marker of recurrence-free rate in non-muscle-invasive bladder cancer, potentially through suppressing cadherin switch, and may act as a potential target for bladder cancer therapy.

     

miR-145 inhibits invasion of bladder cancer cells by targeting PAK1

ObjectivesMicroRNAs play important roles in cancer. In many cancers, miR-145 acts as a tumor suppressor, and it is down-regulated in bladder cancer. In the present study, we explored the modulation of oncogenic gene PAK1 by miR-145 in bladder cancer.Material and methodsExpression of miR-145 was detected in bladder cancer tissues and cell lines by quantitative real-time polymerase chain reaction. Through the bioinformatics approach, PAK1 has been predicted to be a direct target of miR-145 and was confirmed by the PAK1 messenger RNA 3′-untranslated region luciferase activity assay. To investigate whether miR-145 regulates PAK1 expression, it was overexpressed in J82 and T24 bladder cancer cells. In 10 paired bladder normal and tumor tissues, we determined the relationship between miR-145 and PAK1 through quantitative real-time polymerase chain reaction and western blot. By using transwell invasion assay and western blotting analysis, we investigated the effects of miR-145 and PAK1 on bladder cancer cell invasion and expression of invasion marker genes.ResultsThe level of miR-145 decreases and PAK1 protein expression up-regulates in bladder cancer tissue, as compared with the paired normal bladder tissue. Moreover, miR-145 directly targets PAK1 in bladder cancer cells. The level of miR-145 negatively correlates with PAK1 protein expression in bladder cancer. In addition, PAK1 promotes invasion and enhances the expression and activity of MMP-9, whereas miR-145 inhibits bladder cancer cell invasion and expressions of PAK1 and MMP-9.ConclusionsOur results indicate that miR-145 inhibits bladder cancer cell invasion, at least partly through targeting PAK1. Restoration or replacement of miR-145 could be an efficient approach to inhibit PAK1 and bladder cancer development in the tumor therapy.