Cyclin E and p27 protein content in human renal cell carcinoma: clinical outcome and associations with cyclin D

Aberrations in the G1-S transition have been observed in several malignancies, suggesting that cell cycle defects are linked to the activation of oncogenes and inactivation of suppressor genes involved in the transformation process. The frequency of G1/S aberrations in human renal cell carcinoma (RCC) has not been fully clarified. We have therefore analyzed the cyclin E content, using Western blotting, in 79 RCC and 12 corresponding kidney cortex tissues as well as the fraction of p27-positive cells in 73 RCCs, using immunohistochemistry. Most of the tumors (65%) exhibited higher cyclin E levels than corresponding normal kidney cortex tissues. However, only a small fraction of the tumors (3 of 80) had excessive levels of cyclin E when cyclin E levels were compared with proliferation. Cyclin E levels higher than the median value were associated with aneuploidy (p = 0.025), high stage (p = 0.027), high grade (p = 0.013) and high erythrocyte sedimentation rate (ESR; p = 0.005). Cyclin E was further inversely correlated with cyclin D1 (p = 0.023) and positively correlated with cyclin D3 (p = 0.003). Most tumors (76%) demonstrated a normal fraction of p27-positive cells. There was an inverse correlation between p27 positivity and tumor size (p = 0.007), despite a lack of correlation between p27 and proliferation. Patients with p27 low tumors had a poor survival (p = 0.002). There was no correlation between p27 and cyclin E levels. In summary, the results suggest that protein expression of cyclin E and/or p27 is linked to tumor behavior.     

cyclin D1和kip1在肾细胞癌组织中的表达

目的：采用免疫组化方法检测cyclin D1和kip1在肾细胞癌中的表达,以期获得评估肾细胞癌恶性程度及预后评价的指标.方法：将肾细胞癌肿瘤组织及正常组织用免疫组化方法进行染色,按半定量方法进行结果判定,结合临床资料进行分析.结果：cyclin D1和kip1在肿瘤组织与正常组织中的表达具有显著性差异（P＜0.01）.cyclin D1和kip1在不同性别组、年龄组和直径组肿瘤患者组织中的表达对比不具有显著性差异（P＞0.05）.cyclin D1和kip1在不同病理分级组肿瘤患者组织中的表达具有显著性差异（P＜0.01）.kip1表达与肿瘤分化程度呈正相关（r=0.40）.cyclin D1表达与肿瘤分化程度呈负相关（r=0.45）.结论：cyclin D1和kip1在肿瘤细胞的表达呈现一定规律,可以作为判断肾细胞癌肿瘤细胞分化程度及预后的重要指标,同时也为靶向治疗提供了更多的靶点选择.     

p27和cyclin E蛋白表达与膀胱移行细胞癌生物学行为的关系

目的　探讨细胞周期蛋白p2 7和cyclinE表达与膀胱移行细胞癌生物学行为的关系。 方法　对 12 1例膀胱移行细胞癌进行组织病理学分级及分期 ,另取 10例正常膀胱组织作对照 ,采用免疫组织化学SABC法检测各例组织中 p2 7和cyclinE蛋白表达水平 ,并分析其表达与临床病理特征的关系。结果　p2 7和cyclinE蛋白阳性表达率分别为 42 .1% ( 5 1/12 1)和 3 4.7% ( 4 2 /12 1)。p2 7和cyclinE蛋白表达均与肿瘤的组织学分级显著相关 ,但与病理分期无关。p2 7蛋白与cyclinE蛋白表达呈负相关 (P<0 .0 5 )。p2 7阴性组中 ,cyclinE阳性患者 5年生存率低于cyclinE阴性者 (P <0 .0 5 ) ;p2 7阳性组中 ,cyclinE表达状况与患者 5年生存率无显著相关。结论　p2 7和cyclinE是评估膀胱移行细胞癌预后的有价值的指标     

Prognostic implication of cyclin E expression and its relationship with cyclin D1 and p27Kip1 expression on tissue microarrays of node negative breast cancer

BACKGROUND AND OBJECTIVES: Altered expression of cell-cycle regulators is prevalent in clinical breast cancer. This study was performed to analyze the impact of cyclin E expression to the outcome of breast cancer together with cyclin D1 and p27Kip1. METHODS: The correlation between cyclin D1/E and p27Kip1 expression was analyzed in tissue arrays of 175 node-negative breast cancers treated by the same chemotherapy composed of fluorouracil, cyclophosphamide, and methotrexate. Data from the immunohistochemical assays of three molecules were correlated and were analyzed with clinical outcome of the patients. RESULTS: Cyclin E expression was observed in 48 (27.4%) of 175 breast carcinomas. Cyclin E expression was significantly increased in young age patients and poorly differentiate tumors. Expression of cyclin E was significantly increased in cyclin D1 expressing tumors (P = 0.034). p27Kip1 expression was preserved above the 50% level in 87 tumors (49.7%) and was inversely correlated with cyclin E expression (P = 0.042). Ki67 labeling index was significantly increased in cyclin E-expressing tumors (P = 0.033) and was inversely related with p27Kip1 expression. In multivariate survival analysis, cyclin E expression was significant for the prediction of poor survival of the patients. CONCLUSIONS: Cyclin E expression was associated with poor prognosis and intimately correlated with the expression of cyclin D1 and p27Kip1. Integration of TMA technology allowed a high-throughput analysis for correlating molecular in situ findings with clinico-pathologic information.     

应用组织微阵列技术研究细胞周期调控因子p27^kip1和p16^Ink4a在宫颈腺癌中的表达

目的 研究细胞周期调控因子p27^kip1、p16^Ink4a在宫颈腺中的表达及临床意义。方法 应用组织微阵列技术结合免疫组化（二步法）检测106例宫颈腺癌组织和22例慢性宫颈炎组织中p27^kip1和p16^Ink4a的表达。结果 宫颈腺癌组织中p27^kip1蛋白阳性表达率为58．5％，明显低于慢性宫颈炎组织（P〈0．05），p16^Ink4a在宫颈腺癌组织中的阳性表达率为70．8％，明显高于慢性宫颈炎组织（P〈0．01）；p16^Ink4a蛋白表达与宫颈腺癌病理分级和组织学类型无明显相关性；p27^kip1蛋白表达与官颈腺癌病理分级无明显相关性，而与组织学类型相关，透明细胞腺癌p27^kip1阳性表达率明显低于其它类型宫颈腺癌（P〈0．05）。结论 p27^kip1和p16^Ink4a在宫颈腺癌的癌变发生过程中起重要作用，二者可能作为宫颈腺癌恶性化的参考指标。     

Annexin-1在肾细胞癌中的表达

目的：探讨肾细胞癌（renal cell carcinoma tissue,RCC）组织中Annexin-1的表达及意义。方法：应用免疫组织化学方法检测56例肾癌组织、20例癌旁组织和16例肾细胞组织中Annexin-1的表达情况。结果：肾癌组织、癌旁组织和肾细胞组织中Annexin-1的阳性表达率分别为21.43%（12/56）、35%（7/20）和81.25%（13/16）。结论：Annexin-1在肾癌组织、癌旁组织中的表达较肾细胞组织中的表达显著降低,检测Annexin-1有望为肾细胞癌的早期诊治提供理论依据。     

Annexin-1在肾细胞癌中的表达

目的:探讨肾细胞癌(renal cell carcinoma tissue,RCC)组织中Annexin-1的表达及意义。方法:应用免疫组织化学方法检测56例肾癌组织、20例癌旁组织和16例肾细胞组织中Annexin-1的表达情况。结果:肾癌组织、癌旁组织和肾细胞组织中Annexin-1的阳性表达率分别为21.43%(12/56)、35%(7/20)和81.25%(13/16)。结论:Annexin-1在肾癌组织、癌旁组织中的表达较肾细胞组织中的表达显著降低,检测Annexin-1有望为肾细胞癌的早期诊治提供理论依据。     

Expression of miR-27a-3p is an independent predictive factor for recurrence in clear cell renal cell carcinoma

MicroRNAs (miRNAs) are noncoding RNAs that regulate gene expression and function in tumor development and progression. We previously identified up-regulated miRNAs in clear cell renal cell carcinoma (ccRCC) compared to matched-pair normal kidney by microarray. Here, we identify miRNAs that are up-regulated in ccRCC and are also correlated with survival and/or recurrence. Twenty-four samples from ccRCC patients who underwent nephrectomies between 2011 and 2012 were divided into two groups: one of eleven patients who experienced recurrence (Group 1), and one of thirteen patients with no evidence of disease (Group 2) 2 years after surgery. Analyzing 22 miRNAs that were up-regulated in ccRCC in our previous study, we identify five miRNAs that were statistically up-regulated in Group 1 versus Group 2 by quantitative real-time PCR. We then evaluated these miRNAs in an independent cohort of 159 frozen ccRCC samples. High levels of miR-27a-3p (p < 0.01) correlated with a worse progression-free survival rate. Multivariate analysis revealed that miR-27a-3p was an independent predictive factor for recurrence. For functional analysis, miR-27a-3p controlled cell proliferation, migration and invasion in RCC cell lines. MiR-27a-3p could act as oncogenic miRNA and may be a candidate for targeted molecular therapy in ccRCC.     

p27~(Kip1)和细胞周期蛋白D1在胃癌中的表达及其预后意义

目的 :研究p2 7Kip1、细胞周期蛋白D1(cyclinD1)在胃癌组织中的表达水平以及与生物学行为的关系和对预后评价的意义。方法 :以免疫组化方法检测 92例胃癌组织中p2 7Kip1、cyclinD1蛋白的表达水平。结果 :本组 92例胃癌中 ,p2 7Kip1蛋白阳性 39例 ,占 42 .4% ;cyclinD1蛋白表达阳性 44例 ,占 47.8% ;胃癌组织中 ,p2 7Kip1蛋白水平与胃壁浸润深度、TNM分期、病理组织学分级、区域淋巴结转移均相关 (P <0 .0 5 ) ;cyclinD1蛋白表达与病理组织学分级负相关 (P <0 .0 5 ) ;p2 7Kip1与cyclinD1蛋白阳性表达显著相关 (P <0 .0 5 ) ;单变量生存分析结果 ,p2 7Kip1高表达组三年、五年生存率分别为 77.1%、5 7.8% ,明显高于低表达组的 33.7%、2 6 .3 % (P =0 .0 0 7) ,多变量分析显示p2 7Kip1是一个独立的预后指标 (P =0 .0 0 0 3)。结论 :p2 7Kip1可作为反映肿瘤恶性表型的指标 ,对胃癌预后具有一定的价值 ;cyclinD1是胃癌发生、发展过程中早期的分子事件 ;p2 7Kip1在胃癌进展中起着比cyclinD1更重要的作用。     

Cyclin E和p27~(kip1)在膀胱移行细胞癌中的表达及其意义

目的　探讨CyclinE和p2 7kip1在膀胱移行细胞癌 (BTCC)中的表达 ,并评价其相关性和意义。方法　应用鼠抗人CyclinE和 p2 7kip1单克隆抗体对 65例BTCC和 12例正常粘膜进行免疫组化SP法染色。结果　CyclinE在BTCC中阳性表达率随着病理分级 (P <0 .0 5 )和临床分期 (P <0 .0 1)的升高而增高 ,有淋巴结转移组非常显著高于无淋巴结转移组 (P <0 .0 1) ;而p2 7kip1在BTCC中的阳性表达率随着临床分期和病理分级的增高而降低 (P <0 .0 5 ) ,有淋巴结转移组显著低于无淋巴结转移组 (P <0 .0 5 )。CyclinE和 p2 7kip1在BTCC中表达具有负相关性 (r =-0 .2 62 ,P <0 .0 5 )。结论　CyclinE对BTCC的分化、增殖、浸润和转移可能起促进作用 ;而p2 7kip1则对BTCC的分化、增殖、浸润和转移可能起抑制作用 :两者在细胞周期进展中的共同表达失调 ,可能是导致BTCC发生发展和向恶性表型转化的机制之一。     

细胞周期蛋白cyclin D1、CDK4在食管癌中的表达及其意义

目的：获得食管癌发生过程中调节Ｇ１细胞周期各种因子的作用。方法：采用抗ｃｙｃｌｉｎＤ１和ＣＤＫ４的单克隆抗体对１０例下沉食管和５０例食管鳞状上皮癌标本进行免疫组织化学染色。结果：ｃｙｃｌｉｎＤ１和ＣＤＫ４在正常食管上皮呈现较低水平的表达,在食管鳞状上皮癌中则过表达。２７／５０食管鳞状上皮癌ｙｃｙｃｌｉｎＤ１染色阳性,其中８例强阳性,１２例只表达ＣＤＫ４,１１例只表达ｃｙｃｌｉｎＤ１,１４例既表达ｃ     

口腔鳞癌中活化细胞外信号调节激酶1/2和cyclin D1的表达

ras／raf/丝裂原活化蛋白激酶（mitogen-activated protein kinase,MAPK）级联是细胞增殖信号的关键。MAPK信号通路位于ras基因下游,通过激活cyclin D1基因刺激细胞增殖,从而引起恶性肿瘤发生。细胞外信号调节激酶（extracellular signal-regulated kinase,ERK）是MAPK家族的成员之一,包括ERK1和ERK2。ERK1／2被活化及cyclin D1过度表达与肿瘤的发生有关。我们应用免疫组化方法,检测了活化ERK1／2和cyclin D1蛋白在口腔鳞癌及相应癌旁正常组织中的表达,以探讨其与口腔鳞癌临床生物学特性和细胞增殖的关系,并同时检测了作为细胞增殖因子的Ki-67。     

Alteration of subcellular and cellular expression patterns of cyclin B1 in renal cell carcinoma is significantly related to clinical progression and survival of patients

Cyclin B1, identified as a regulator of late cell cycle, is involved in the development and progression of a variety of human malignancies. To clarify the role of cyclin B1 in the pathogenesis and prognosis of renal cell carcinoma (RCC), protein expression was compared with clinicopathological characteristics of patients as well as the long-term survival after surgical therapy. Expression analysis was carried out by immunohistochemistry and tissue microarray analysis. The microarrays that represented the primary tumors, their invasion front and normal peritumoral renal parenchyma contained 753 tissue cores obtained from 251 randomly selected nephrectomy specimens. Immunopositivity within the primary tumors was significantly associated with tumor stage (pT) (p < 0.01), lymph node status (pN) (p < 0.01) as well as the presence of systemic metastatic disease (p = 0.01). Subcellular expression in the cytoplasm of tumor cells significantly correlated with pT (p = 0.02) and pN (p = 0.03). When peritumoral tissue samples exhibited a relative amount of <10% of positively reacting epithelial cells, cyclin B positivity was identified to predict long-term survival of patients in univariate analysis (p < 0.01) whereas borderline significance was observed in multivariate statistical analysis (p = 0.05). Increased intratumoral cyclin B1 positivity and aberrant localization of signals within the cytoplasm of tumor cells is positively correlated with the tendency towards tumor progression, indicating the significant role of cyclin B1 in the development and pathogenesis of RCC. The result of uni- and multivariate statistical analysis suggests the prognostic value of cyclin B1 for RCC patients.     

MiR-210 regulates cell cycle in nasopharyngeal carcinoma cell line （CNE-1） under hypoxic condition by reducing the expression of cyclin DI

Objective： The aim of our study was to determine the underlying mechanism of miR-210 on regulation of the cell cycle in nasopharyngeal carcinoma cell line CNE-1, particularly through regulation of cyclin D1, under hypoxic conditions. Methods： The CNE-1 cell line was induced with hypoxia, and the expression levels of endogenic miR-210 and cyclin D1 were detected by real-time PCR and Western blotting. Next, the luciferase assay was used to confirm that cyclin D1 is a target gene for miR-210. Cell cycle and cell proliferation were detected in CNE-1 cells that were cultured under hypoxic conditions with either overexpression or knockout of miR-210 using flow cytometry and MTT assay, respectively. Results： Hypoxia induced the expression of miR-210, resulting in reduced mRNA and protein levels of cyclin D1 and repression of cyclin D1 in CNE-1 cells. Further analysis indicated that miR-210 directly binded to the 3＇UTR of the cyclin D1 gene, thus regulated the expression of cyclin DI. The flow cytometry assay showed that, under hypoxic conditions, miR-210 blocked CNE-1 cells in the G1 phase, and miR-210 also inhibited the proliferation of CNE-1 cells. Conclusion： Under hypoxic conditions, miR-210 directly reduced the expression of cyclin D1, leading to CNE-1 cells blocked in G1 phase.     

Retinoblastoma protein in human renal cell carcinoma in relation to alterations in G1/S regulatory proteins

The retinoblastoma gene product (pRb) is the main substrate for cyclin-dependent kinases (CDKs) during the G1/S transition. Aberrations in cell cycle regulatory proteins, which have been observed in many malignancies, can theoretically cause increased phosphorylation of pRb due to unbalanced CDK activities. The expression and phosphorylation of pRb and potential associations to cell cycle aberrations in renal cell carcinomas (RCC) has only partly been clarified. We therefore evaluated the presence of pRb and the level of pRb-phosphorylation in 216 RCCs arranged in tissue microarrays by using different pRb-antibodies, including pRb-phosphospecific antibodies. Most RCCs (95%) expressed pRb, while cases with the low pRb levels, potentially indicative for pRb-inactivation, were few. In order to detect secondary alterations to a potential pRb-inactivation, the p16 expression was also monitored. None of the tumors exhibited increased p16 levels, confirming that pRb-inactivation is rare in RCC. Phosphorylated pRb was detected in approximately 50% of the RCCs, using Western blotting or immunohistochemistry. The immunohistochemical ppRb(ser807/811) levels were associated with high proliferation, cyclin D1, cyclin E and p27 protein content. Surprisingly, there was no association between pRb-phosphorylation and clinicopathological data. In summary, pRb seemed to be functional and aberrations in G1/S-regulatory proteins were associated with increased phosphorylation of pRb and proliferation. The data supports that pRb might be one of the main cell cycle regulators in RCC.     

小鼠前胃癌组织形成中VEGF和cyclin D1及p27^kip1的表达

目的：研究VEGF、cyclin D1和p27^kip1在小鼠前胃癌组织形成过程中的表达及意义，初步探讨维甲酸对小鼠前胃癌的干预机制。方法：建立ICR小鼠前胃癌模型，SABC法测定VEGF、cyclin D1和p27^kip1表达。结果：cyclin D1在小鼠前胃癌组织中的表达（64．7％）明显高于癌前期病变（29．4％）。X^2=4．3447，P=0．0371；VEGF表达在癌前期病变（47．1％）高于增生期病变（33．3％），X^2=4．836，P=0．0381；cyclin D1、p27^kip1和VEGF阳性表达与小鼠胃癌组织学分级无相关性。维甲酸抑制组癌及癌前期病变发生率（23．5％）低于单纯诱导组（76．9％），X^2=14．3549．P=0．0002。结论：VEGF、cyclin D1和p27^kip1是小鼠前胃癌组织形成过程中重要的分子事件；维甲酸对亚硝胺类致癌物诱发的小鼠前胃鳞状上皮癌及癌前期病变具有抑制作用，但与对VEGF、cyclin D1和p27^kip1表达的调控无明显相关。     

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.     

组织芯片应用于原发性鼻咽癌中p53表达的高通量分析

目的 :通过对不同临床分期鼻咽癌(NPC)中 p5 3蛋白表达的进一步研究 ,探讨高通量组织微阵列技术 (high throughputtis suemicroarray)应用于大规模临床标本高效筛查的可行性。方法 :用组织阵列仪 (tissuearrayer)制备连续系列分区排列的组织芯片 ,然后用 p5 3单抗免疫组织化学法检测一张组织芯片上 3 2 0例各类型的鼻咽活检组织标本中 p5 3蛋白表达 ,并分析 p5 3表达在不同鼻咽组织的分布状况及其与临床分期的相关性。结果 :在正常鼻咽黏膜组织、癌旁组织及癌组织中 p5 3蛋白的检出率分别为 0 ( 0 / 2 9)、16 7% ( 4 / 2 4)及 61 4% ( 164 /2 67) ,NPC组织中 p5 3蛋白表达率明显高于其他鼻咽非癌组织 ,P <0 0 0 1,其中癌旁 4例全为弱阳性 ,癌组织中 164例中弱阳性为112例 ,占 41 9% ( 112 / 2 67) ,而强阳性为 5 2例 ,占 19 5 % ( 5 2 / 2 67) ;p5 3表达与鼻咽癌的临床分期无显著相关性 ,P =0 0 78,而p5 3过表达 (强阳性 )却与鼻咽癌的临床中晚期密切相关 ,P =0 0 0 71。结论 :鼻咽癌中 p5 3蛋白的高频核积累提示p5 3蛋白功能的抑制在鼻咽癌的发生发展中起重要作用 ;应用组织芯片 (tissuechip)大规模高效筛查临床组织标本是可行性的 ,具有快速、方便、经济的特点     

应用组织芯片技术研究人类胰腺癌中PSCA的表达

目的检测PSCA在胰腺癌中的表达情况,探讨PSCA在胰腺癌发病中所起的作用。方法用组织芯片技术构建包含78例导管腺癌,12例慢性胰腺炎病人,10例正常胰腺组织的100点阵的石蜡组织芯片。用免疫组化SP法检测该芯片中PSCA的表达,分析其与胰腺癌临床病理因素的关系。结果78例胰腺癌病人中,PSCA阳性表达率为79．5％（62／78）,与正常组比较PSCA表达与胰腺癌显著相关（χ^2=15．81,P〈0．005）,与慢性胰腺炎比较PSCA亦与胰腺癌显著相关（χ^2=11．33,P〈0．005）;PSCA表达与年龄、性别、组织分化程度及TNM分期无明显相关性。结论PSCA阳性表达与胰腺癌相关,可能与胰腺癌的发生发展有着密切关系,但与胰腺癌的临床病理特型无关。     

胃癌中细胞周期素E蛋白的表达及意义

Objective  

The aim of the study was to investigate the expression and significance of cyclin E in gastric carcinoma.