Shank1 在肾细胞癌组织中的表达及临床意义

目的:检测Shank 家族成员Shank1 在肾细胞癌（renal cell carcinoma,RCC ）中的表达,探讨在RCC 癌组织与癌旁组织中Shank1 的表达差异,分析其与RCC 临床病理特征的关系。方法:收集2008年5 月至2014年12月沧州市中心医院与济南市中心医院120 例RCC 术后患者的癌组织与癌旁组织标本,采用免疫组织化学染色及Westernblot检测Shank1 的蛋白表达水平,分析Shank1 表达与RCC 临床病理特征的关系。结果:免疫组织化学结果显示,RCC 的癌组织中Shank1 表达较癌旁组织明显上调,差异具有统计学意义（P < 0.05）。 Westernblot检测发现RCC 的癌组织中Shank1 蛋白表达水平明显高于癌旁组织。对Shank1 表达上调RCC 患者的临床病理特征分析发现,Shank1 在癌组织中的高表达与患者的性别、年龄、肿瘤的大小、TNM 分期无显著性相关（P > 0.05）,但与RCC 的不同病理类型显著性相关（P < 0.05）。 结论:Shank1 在RCC 的癌组织中异常表达,并与RCC 的病理类型相关。      

Dishevelled2基因及其蛋白在透明细胞性肾细胞癌组织中的表达及其临床意义

目的探讨透明细胞性肾细胞癌（CCRCC）组织及其癌旁组织中Dishevelled2（DVL2）基因及其蛋白的表达情况及其临床意义。方法用半定量反转录一聚合酶链反应（RT—PCR）和荧光实时定量PCR方法检测22例CCRCC患者的原位肾癌组织及其配对癌旁组织DVL2基因表达情况,并检测了32例总样本中Ⅲ＋Ⅳ期与I＋Ⅱ期肾癌组织中DVL2基因的表达差异;应用免疫组织化学法回顾性研究22例CCRCC患者的原位肾癌组织及其配对癌旁组织中DVL2蛋白的表达情况,同时检测了10例无正常肾组织配对的CCRCC患者肿瘤组织。结果半定量RT—PCR结果表明,在22例CCRCC样本中,DVL2mRNA在17例（77．2％）CCRCC原发灶中的表达较其配对癌旁组织上调,其结果与实时荧光定量PCR结果基本一致（仁2．535,P=0．0197）,实时荧光定量PCR结果显示,13例Ⅲ＋Ⅳ期CCRCC中8例（61．5％）肾癌组织DVL2mRNA表达高于I＋Ⅱ期。肾癌组织;免疫组织化学分析表明,DVL2蛋白定位于CCRCC细胞的胞膜及胞质,22例CCRCC中有18例（81．8％）癌组织表达强度高于癌旁组织,但32例CCRCC的DVL2蛋白表达在不同年龄、性别、肿瘤分期间差异均无统计学意义（Fisher精确概率法,均P〉0．05）。结论在mRNA及蛋白水平上,CCRCC中DVL2表达水平显著高于癌旁组织,并且Ⅲ＋Ⅳ期肿瘤患者原位癌组织中DVL2的表达量高于I＋Ⅱ期,提示DVL2可能是一个潜在的与CCRCC发生及转移相关的分子标志物。     

Tenascin-C在肾透明细胞癌中的表达及意义

目的:分析Tenascin-C（TN-C）在肾透明细胞癌中的表达及其与临床病理因素间的关系。方法:应用免疫组化方法检测肾透明细胞癌中TN-C蛋白表达情况。结果:免疫组化结果显示40.63%（39/96）的病例存在肿瘤间质TN-C蛋白的过表达,且过表达与肿瘤组织核分级负相关（P=0.022）;18.75%（18/96）的病例存在肿瘤细胞胞质TN-C蛋白的过表达,且过表达与肿瘤组织核分级、p-TNM分期正相关,分别为（P＜0.001;P=0.038）。肿瘤间质及肿瘤细胞胞质的过表达与RCCC患者的年龄、性别、肿瘤大小均无明显相关性。结论:肾透明细胞癌中存在TN-C蛋白肿瘤间质及肿瘤细胞的过表达,并与肿瘤分化和p-TNM分期相关。     

Vimentin在肾透明细胞癌中的表达及临床意义

目的:探讨波形蛋白(Vimentin)在人肾透明细胞癌(clear cell renal cell carcinoma,ccRCC)和癌旁组织样本中的表达情况,并分析其与 ccRCC 临床分期、病理分级和侵袭转移等恶性生物学特性的关系及临床意义。方法:Western blot方法检测29例新鲜肾透明细胞癌及相应癌旁组织中Vimentin的表达;免疫组织化学方法检测120例肾透明细胞癌及相应癌旁组织中Vimentin的表达情况,分析Vimentin在肾透明细胞癌组织中及癌旁正常肾脏组织中的表达水平差异及Vimentin的表达与肿瘤大小、临床分期、组织病理分级、远处转移的关系。结果:Vimentin在肾透明细胞癌及相应癌旁组织中均有表达,前者的表达水平较后者显著升高（P＜0.01）;免疫组织化学显示肾透明细胞癌组织中,Vimentin的阳性表达率高达90.0%,与癌旁组织存在明显差异（P＜0.01）;Vimentin在肾透明细胞癌组织中以胞膜表达为主,其表达随肾癌的临床分期、肾包膜受侵、癌栓形成或远处转移有升高趋势,但组间比较经统计学分析均无显著性差异。结论:Vimentin在肾透明细胞癌组织中的表达明显高于癌旁正常肾组织,提示Vimentin在肾透明细胞癌的发生发展过程中发挥重要的作用。Vimentin在肾透明细胞癌组织中过表达,与肾癌的发展密切相关,可以作为肾细胞癌早期诊断、预后和复发评估的参考指标之一。     

CD248在肾透明细胞癌中的表达及临床意义CSCD

目的探讨内皮唾酸蛋白(CD248)在肾透明细胞癌(ccRCC)组织中的表达及临床意义。方法收集115例ccRCC组织及对应癌旁组织的石蜡标本,同时收集其中17例ccRCC组织及对应癌旁组织的新鲜标本。qRT-PCR检测17例ccRCC组织及对应癌旁组织中CD248 mRNA的表达;免疫组织化学法检测115例ccRCC患者石蜡标本的CD248蛋白水平,分析CD248表达与ccRCC临床病理指标及预后的相关性。结果ccRCC组织中的CD248阳性表达免疫组织化学反应强度显著强于癌旁组织,差异有统计学意义(P=0.0061),且CD248蛋白阳性信号定位于细胞核及细胞质。CD248 mRNA在癌旁组织中的表达水平明显低于ccRCC组织(P=0.0026)。CD248高表达组组间总生存率明显低于CD248低表达组(44.4 vs.57.2月,P=0.017)。结论CD248的表达可能参与ccRCC的发生发展。与癌旁组织比较,CD248在ccRCC组织中高表达;CD248高表达的ccRCC患者术后生存时间较短。     

膜细胞骨架蛋白-ezrin在肾细胞癌中的表达及意义

 目的 从蛋白和转录水平观察肾癌组织中ezrin蛋白的表达及其与肾癌临床病理特征的关系。 方法 收集80例肾癌组织及40例癌旁正常肾组织,应用组织芯片和免疫组化检测ezrin蛋白的表达情况;同时取其中50例肾癌组织及40例癌旁正常肾组织行RT-PCR检测其mRNA表达。 结果 ezrin蛋白的表达定位于细胞浆和细胞膜。ezrin蛋白的阳性表达率在肾癌组织中为95%(76/80),显著高于正常肾组织的阳性表达率75%(30 /40)(χ²=10.350,P=0.002);ezrin mRNA表达阳性率在肾癌组织中为82%（41/50),显著高于正常肾组织62.5%(25/40)(χ²=4.321,P=0.038)。免疫组化评分及ezrin mRNA阳性表达率淋巴结转移组显著高于无淋巴结转移组[(9.500±2.502) vs. （7.232±3.045）, 13/14 vs. 16/36,P<0.05];腔静脉瘤栓组显著高于无腔静脉瘤栓组[(9.315±3.027） vs. （7.563±2.991）, 14/14 vs. 22/36,P<0.05]。而肾癌组织中ezrin蛋白, ezrin mRNA表达水平与患者的性别、年龄、肿瘤分期、分级、肿瘤大小及肿瘤病理类型无关(P>0.05)。 结论 肾癌组织中ezrin蛋白及mRNA水平均呈高表达,其表达的强度与肿瘤淋巴结转移,腔静脉瘤栓形成有关。ezrin可能参与了肾癌的发生、侵袭和转移,有望成为肾细胞癌生物学行为的评估指标。     

Notch3在肾透明细胞癌中的表达及临床意义

目的:通过研究Notch3蛋白在肾透明细胞癌组织和癌旁组织中的表达变化情况,探讨其临床意义及相关影响因素。方法:采用免疫组织化学染色检测57例肾透明细胞癌组织、38例癌组织外缘1.0 cm处组织中的Notch3蛋白表达情况,并观察其与临床病理参数之间的关系。结果:肾透明细胞癌、癌旁组织中Notch3蛋白的阳性表达率分别为80.7%（46/57）、31.6%（12/38）,P＜0.05。单因素分析表明,Notch3蛋白的表达与肾透明细胞癌的临床分期、病理学分级、远处转移密切相关（P＜0.05）。多因素分析表明,患者的年龄、临床分期、吸烟史是Notch3蛋白在肾透明细胞癌中表达的独立危险因素（P＜0.05）。结论:Notch3蛋白在肾透明细胞癌组织中的表达明显增高,Notch3可能成为肾透明细胞癌的潜在基因治疗靶点和早期诊断检测及预防的指标之一。     

迁移侵袭抑制蛋白在肿瘤发生发展中的作用及其临床意义

迁移侵袭抑制蛋白（migration and invasion inhibitory protein,MIIP）能够与胰岛素样生长因子结合蛋白2（insulin-like growth factor binding protein 2,IGFBP2）、组蛋白去乙酰化酶6（histone deacetylase 6,HDAC6）、p21活化激酶1、表皮生长因子受体、细胞分裂周期20（cell division cycle 20,cdc20）、拓扑异构酶等相互作用,抑制细胞增殖和迁移侵袭,进而抑制多种肿瘤的发生发展。最近的研究显示,MIIP还与病毒感染、细胞免疫有关。鉴于MIIP的作用涉及到多条肿瘤相关的信号通路及肿瘤治疗靶点,MIIP逐渐成为研究的热点。本文主要围绕MIIP的分子特征、功能和在多种肿瘤中的临床意义及作用机制进行综述。      

细胞周期蛋白D1和E在食管鳞状细胞癌中的表达及意义

目的　探讨细胞周期蛋白 D1 和 E( cyclin D1 and cyclin E)在食管鳞状上皮癌中的表达及意义。方法　采用 S- P免疫组化方法 ,检测了周期蛋白 D1 和 E在正常食管上皮 ( 2 0例 )、增生上皮 ( 2 1例 )和鳞癌 ( 6 2例 )中的阳性率。结果　正常鳞状上皮中 cyclin E无阳性表达 ,在增生上皮中 cyclin E阳性率为 33.3% ,在鳞癌中阳性率为5 3.2 % ;cyclin D1 在正常鳞状上皮阳性率为 5 .0 % ,在增生和鳞癌中阳性率分别为 4 2 .9%和 4 6 .8% ;cyclin E在低分化鳞癌组阳性率 ( 80 .0 % )明显高于高分化组 ( 2 9.0 % ) ( P<0 .0 5 )。 cyclin D1 和 cyclin E在鳞癌中的阳性表达呈正相关 ( r=0 .782 )。结论　 cyclin D1 和 cyclin E过表达与食管鳞状细胞癌的发生和分化程度有关。     

FHIT基因在食管鳞状细胞癌及癌旁组织中的表达

目的 :探讨FHIT基因在食管鳞状细胞癌及癌旁组织中的表达及意义。方法 :应用免疫组织化学S P法对 6 4例食管鳞状细胞癌组织标本及癌旁组织进行FHIT基因的表达水平检测 ,分析其与临床病理分期、细胞分化程度及淋巴转移等的关系。结果 :食管鳞状细胞癌组织中FHIT基因阳性表达率为 2 8 1% (18/6 4 ) ,癌旁组织中阳性表达率为 87 5 % (5 6 /6 4 ) ,二者差异有显著统计学意义 (χ2 =4 6 3,P <0 0 1) ,FHIT基因表达下降与患者PTNM分期、细胞分化程度、淋巴结转移、性别、年龄、吸烟史、饮酒史及病史长短均无明显关系 (P >0 0 5 )。结论 :FHIT基因的表达下降可能与食管鳞状细胞癌的发生、发展相关 ,与临床病理分期、细胞分化程度及淋巴结转移无关。     

MIIP对肾癌细胞增殖、侵袭及迁移能力的影响

目的:利用质粒转染技术制备过表达迁移侵袭抑制蛋白(migration and invasion inhibitory protein,MIIP)的肾癌786-0/MIIP细胞系,观察MIIP对细胞增殖、侵袭及迁移能力的影响.方法:构建携带MIIP基因的质粒,通过脂质体质粒转染技术上调肾癌786-0细胞系MIIP基因表达.Western-blot、Real-time PCR检测转染效果;利用MTT实验检测MIIP对786-0细胞增殖能力的影响;Transwell实验、划痕实验检测MIIP对786-0细胞侵袭、迁移能力的影响.结果:Westem-blot、Real-time PCR检测发现实验组786-0/MIIP细胞MIIP蛋白及mRNA表达量有效上调.MTT实验显实验组786-0/MIIP细胞增殖能力明显减弱(P＜0.01).Transwell实验显实验组786-0/MIIP细胞侵袭能力减弱,穿透小室细胞数明显减少(P＜0.01).细胞划痕实验显实验组786-0/MIIP细胞迁移能力显著下降(P＜0.01).结论:通过质粒转染技术能够制备过表达MIIP的肾癌细胞系.转染成功的肾癌786-0/MIIP细胞系MIIP表达量增加,有效抑制了肾癌细胞的增殖、侵袭与迁移.     

Regulation of carcinoma cell invasion by protein C inhibitor whose expression is decreased in renal cell carcinoma

Protein C inhibitor (PCI), a member of the serine protease inhibitor family, is produced in various human tissues, including the liver, kidney and testis. In addition to inhibiting the anticoagulant protein C pathway, PCI also inhibits urinary plasminogen activator (uPA), which is a well-known mediator of tumor cell invasion. In the present study, to clarify the biologic significance of PCI in the kidney, we compared the expression of PCI between human renal cell carcinoma (RCC) tissue and nontumor kidney tissue. The PCI antigen level in RCC tissue was found to be significantly lower than in nontumor kidney tissue, and expression of PCI mRNA was detected in normal renal proximal tubular epithelial cells (RPTEC), but not in RCC or in an RCC cell line (Caki-1 cells). No differences were detected between the nucleotide sequence of the major cis-elements in the promoter region of the PCI gene from nontumor kidney and RCC tissues, RPTEC and Caki-1 cells, an RPTEC-derived RCC cell line. The in vitro invasiveness of Caki-1 cells transfected with a PCI expression vector was significantly decreased compared to mock-transfected Caki-1 cells, and it was blocked in the presence of anti-PCI antibody. Since PCI itself did not affect the proliferation rate of Caki-1 cells or cell expression of uPA in vitro, the effect of uPA, PCI, heat-inactivated PCI and plasminogen activator inhibitor (PAI)-1 on the invasive potential of cultured RCC cells was evaluated. The in vitro invasiveness of Caki-1 cells, which express uPA, was significantly enhanced by the addition of uPA, and it was inhibited by anti-uPA antibody, PCI and PAI-1, but not by heat-inactivated PCI. In addition, uPA activity was significantly decreased and uPA-PCI complex level was significantly increased in the culture medium of PCI expression vector-transfected Caki-1 cells as compared to mock-transfected Caki-1 cells. These findings strongly suggest that PCI regulates the invasive potential of RCC cells by inhibiting uPA secreted by these cells. The results of our study suggest that PCI might be a potential therapeutic agent for inhibiting renal tumor invasion.     

下调Ets2表达对肾癌786-0细胞迁移及侵袭的影响

目的:探讨E26转录因子2（E-twenty six 2,Ets2）对肾癌细胞786-0迁移、侵袭的影响及其机制。方法:设计并合成特异性siRNA下调肾癌细胞系786-0中Ets2的表达;并利用划痕实验及Transwell实验检测其迁移及侵袭能力的变化;通过Western blot检测下调Ets2表达后基质金属蛋白酶2（MMP-2）及MMP-9表达水平变化。结果:siRNA能有效下调肾癌786-0细胞系中Ets2的表达。细胞划痕实验结果显示下调Ets2表达后,细胞迁移能力减弱（P＜0.01）;Transwell实验中NC组穿膜细胞数为（43.80±3.99）个,而siRNA1组为（23.30±4.24）个(P＜0.01), siRNA2组为（24.20±3.29）个（P＜0.01）,细胞侵袭能力减弱。Ets2-siRNA介导的Ets2基因沉默下调了786-0细胞中MMP-2及MMP-9的表达的同时上调了基质金属蛋白酶组织抑制剂1（TIMP1）和TIMP2的表达。结论:Ets2可通过调节MMPs及TIMPs的表达影响肾癌细胞的迁移及侵袭能力。     

癌基因Gli2促进肾癌细胞的体外侵袭能力

目的:研究Gli2与肾癌细胞侵袭之间的关系。方法:采用免疫组织化学染色的方法对41例肾癌临床标本中Gli2的表达情况进行检测,并分析Gli2在局部侵袭性肾癌和非局部侵袭性肾癌中的表达差异。运用肾癌细胞模型,通过过表达Gli2质粒或添加Gli2特异性抑制剂干预细胞,采用体外侵袭实验分析Gli2在肾癌细胞侵袭能力中的作用,RT-PCR检测Gli2对细胞侵袭相关基因的调控作用。结果:Gli2蛋白在T3-4期肾癌中的表达水平明显高于T1-2期（P＜0.05）,Gli2过表达能明显促进肾癌细胞的体外侵袭能力,反之,Gli2特异性的抑制剂Gant61则显著减弱肾癌细胞的体外侵袭能力。Gli2抑制肾癌细胞上皮标记物E-Cadherin的表达,上调间质标记物N-Cadherin,Vimentin,MMP-2及MMP-9的表达;Gli2抑制剂的作用则相反。结论:Gli2通过调控侵袭相关基因的表达促进肾癌细胞的侵袭。     

Protein kinase C in human renal cell carcinomas: role in invasion and differential isoenzyme expression

The role of protein kinase C (PKC) in in vitro invasiveness of four different human renal cell carcinoma (RCC) cell lines of the clear cell type was investigated. Different PKC-inhibitors markedly inhibited invasiveness of the highly invasive cell lines, suggesting an invasion-promoting role of PKC in human RCC. Analysis of PKC-isoenzyme expression by protein fractionation and immunoblotting revealed that all cell lines expressed PKC-alpha, -epsilon, -zeta, -mu and -iota as known from normal kidney tissue. Interestingly, PKC-delta, known to be expressed by normal kidney epithelial cells of the rat, was absent on protein and RNA levels in all RCC cell lines investigated and in normal human kidney epithelial cells. PKC-epsilon expression levels correlated positively with a high proliferation activity, but no obvious correlation between expression levels of distinct PKC-isoenzymes and in vitro invasiveness was observed. However, by immunofluorescence microscopy, membrane localisation of PKC-alpha and PKC-epsilon reflecting activation of the enzymes, was associated with a highly invasive potential. In conclusion, our results suggest a role for PKC in invasion of human RCCs and might argue in favour of a particular role of PKC-alpha and PKC-epsilon. Our results further suggest that organ-specific expression patterns of PKC-isoenzymes are not necessarily conserved during evolution.     

MiR-200c-3p inhibits cell migration and invasion of clear cell renal cell carcinoma via regulating SLC6A1

In this study, we investigated the mechanism of miR-200c-3p and SLC6A1 in regulating cell activity of clear cell renal cell carcinoma (CCRCC). The mRNA and miRNA expressions of tissue specimens were analyzed by CapitalBio Corporation (Beijing, China). The expression of SLC6A1 in CCRCC cells was examined through qRT-PCR and western blot. The migration and invasion ability of 786-O cells was testified by transwell assay after transfected. 786-O cell proliferation ability was detected by MTT assay. Dual luciferase reporter assay verified the association between SLC6A1 and miR-200c-3p. SLC6A1 was high expressed and miR-200c-3p was low expressed in CCRCC tissues and cells. Besides, lower SLC6A1 expression indicated longer survival time and higher survival rate. MiR-200c-3p could directly target at SLC6A1 and reduce its expression. MiR-200c-3p inhibited the proliferation, migration and invasion in 786-O cells by down-regulating SLC6A1 expression. The results suggested that the miR-200c-3p served as a suppressor for CCRCC via down-regulating SLC6A1.     

Measurement of progesterone receptor in human renal cell carcinoma and normal renal tissue

Progesterone receptor was measured in eight samples of renal cell carcinoma, nine samples of normal renal tissue, and one sample of melanoma tissue. Progesterone receptor was identified in all samples, with the exception of one renal cell carcinoma. Three patients, all with receptor-positive tumors, were treated with medroxyprogesterone acetate for metastatic disease. In one of these patients there was a partial objective response to treatment. Further research regarding progesterone receptor in renal cell carcinoma is indicated.     

肾透明细胞癌中Spl的表达特征及意义

目的：探讨核转录因子Spl在肾透明细胞癌（renal clear cell carcinoma,RCCC）组织中的表达特征和临床意义。方法：用免疫组织化学PV-6000法,检测59例RCCC和11例正常肾脏组织中Spl的表达,分析其与肿瘤分级和分期的关系。结果：Spl在RCCC组织中阳性表达率为71.2%（42/59）,明显高于正常肾组织的18.2%（2/11）。Sp1蛋白阳性表达与RCCC的组织学分级和临床分期正相关。结论：转录因子Spl可作为一个新的与RCCC发生、发展密切相关的生物学标记物。     

Twist在肾细胞癌中表达及其临床意义研究

目的：研究twist的表达与肾细胞癌（renal cell carcinoma,RCC）恶性进展的关系.方法：应用免疫组织化学及Western blot方法检测twist在RCC及其对应癌旁组织的表达情况,并分析其在肿瘤组织中的表达水平与患者年龄、性别、T分期、Fuhrman分级及淋巴结转移等临床病理资料之间的相关性.结果：twist在RCC阳性表达率为73.1％,显著高于其在对应癌旁组织阳性表达率15.4％ （P ＜0.05）.twist在肿瘤组织的阳性表达率在小于及大于49岁组中分别为69.2％、76.9％;在男、女组中分别为75％、71.9％;T1＋2及T3＋4期分别为61.3％、90.5％;在G1、G2及G3组依次为41.7％、80.8％、85.7％;在无及有淋巴结转移组分别为60％、90.9％.统计学分析提示twist表达差异在T分期、Fuhrman分级及淋巴结转移具有统计学意义（P＜0.05）.Western blot检测发现twist在肾细胞癌组织表达水平显著强于其对应癌旁组织（P＜0.05）.结论：twist在肾细胞癌中高表达与肾癌恶性进展密切相关,有望成为肾癌诊断及治疗新的标记物.     

TRIM44 promotes cell proliferation and migration by inhibiting FRK in renal cell carcinoma

TRIM44 has oncogenic roles in various cancers. However, TRIM44 expression and its function in renal cell carcinoma (RCC) are still unknown. Here in this study, we investigated the clinical significance of TRIM44 and its biological function in RCC. TRIM44 overexpression was significantly associated with clinical M stage, histologic type (clear cell) and presence of lymphatic invasion (P = .047, P = .005, and P = .028, respectively). Moreover, TRIM44 overexpression was significantly associated with poor prognosis in terms of cancer‐specific survival (P = .019). Gain‐of‐function and loss‐of‐function studies using TRIM44 and siTRIM44 transfection showed that TRIM44 promotes cell proliferation and cell migration in two RCC cell lines, Caki1 and 769P. To further investigate the role of TRIM44 in RCC, we performed integrated microarray analysis in Caki1 and 769P cells and explored the data in the Oncomine database. Interestingly, FRK was identified as a promising candidate target gene of TRIM44, which was downregulated in RCC compared with normal renal tissues. We found that cell proliferation was inhibited by TRIM44 knockdown and then recovered by siFRK treatment. Taken together, the present study revealed the association between high expression of TRIM44 and poor prognosis in RCC patients and that TRIM44 promotes cell proliferation by regulating FRK.