Cyclin E、CDK2和p21WAF1在食管上皮癌变过程中的表达及意义

目的探讨食管上皮癌变过程中细胞周期调控因子cyclin E、CDK2和p21WAF1的表达状况及其意义.方法应用免疫组化SP法和原位杂交方法分别检测48例食管癌组织、31例非典型增生组织和17例正常食管粘膜中cyclin E、CDK2和p21WAF1蛋白及mRNA表达.应用半定量RT-PCR和Western blot检测22例新鲜食管癌及相应癌旁组织的mRNA和蛋白表达.结果从食管正常粘膜、非典型增生组织到癌组织,cyclin E和CDK2蛋白和mRNA阳性表达率逐渐上升,差异具有统计学意义(P＜0.01或P＜0.05).食管癌组织中cyclin E、CDK2和p21WAF1蛋白及mRNA高表达,与癌旁组织或切缘正常食管粘膜有显著性差异(P＜0.01).cyclin E、CDK2和p21WAF1基因表达显著正相关(P＜0.01或P＜0.05).结论食管上皮癌变过程中,细胞周期相关基因cyclin E和CDK2表达逐渐增强.cyclin E基因表达异常是食管癌变过程中的早期事件.p21WAF1基因在食管癌中高表达,可能与细胞周期调控的反馈机制有关.     

口腔鳞状细胞癌细胞周期因子p27 cyclin D1及CDK4的表达与意义

目的：从细胞周期调控的角度来探讨口腔鳞状细胞癌（OSCC）的发生、发展及预后和p27、cyclin D1和CDK4的关系。方法：采用免疫组织化学方法，检测了50例口腔鳞状细胞癌及10例正常口腔粘膜中p27、cyclinD1和CDK4蛋白表达的水平，然后用Spearman 相关分析检查它们与临床病理学指标的关系以及它们三者之间的相关关系，用Kaplan-Meier方法绘制生存曲线并进行生存分析，Cox比例风险模型作预后分析。结果：1)p27在所有正常口腔粘膜上皮均呈现高表达，而在口腔鳞状细胞癌组织表达降低，p27的低表达与临床分期，淋巴结转移有显著性相关关系，cyclin D1和CDK4在正常粘膜上皮呈现低水平表达，在口腔钙状细胞癌组织中过表达。2)cyclinD1和CDK4在正常粘膜上皮呈现低水平表达，在口腔鳞状细胞癌组织中过表达.2)cyclin D1的表达与CDK4呈正相关（r=0.442,P=0.001）;p27与CDK4的表达呈负相关（r=-0.384,P=0.006）.3)Kaplan-Meier生存分析显示p27高表达组（ ）的各期的生存均高于低表达组（-）,cyclin D1染色（ ）组的生存率低于（-）组。4）Cox比例风险模型分析结果显示p27蛋白表达水平，cyclinD1蛋白表达水平，淋巴结转移和临床分期分别是口腔鳞状细胞癌的独立预后指标。结论：口腔鳞状细胞癌组织中，p27,cyclin D1和CDK4蛋白的异常表达说明它们均参与了口腔鳞状细胞癌的发生发展，且在这一过程中三者之间存在相互协同与制约关系。在临床应用中有可能将p27和cyclin D1蛋白的表达程度作为判断口腔鳞状癌患者预后的指标。     

胃癌中p16,p27kip1和CDK4表达的比较及其意义

目的 研究胃癌组织中p16、p27^kip1蛋白和细胞周期依赖性激酶4（cyclin—dependent kinase4，CDK4）的表达，探讨它们与胃癌临床病理特征的关系及其对预后的影响。方法 应用免疫组化S-P法，检测120例胃癌，20例胃粘膜非典型增生和30例正常胃粘膜组织切片中p16和p27^kip1和CDK4、PCNA的表达，分析它们与临床病理参数的关系。结果 120例胃癌中p16，p27^kip1和CDK4，PCNA蛋白的表达率分别为43．3％，52．5％和46．7％，70．0％，与正常胃粘膜和非典型增生相比有显著性差异（P〈0．01）。p16和p27^kip1的表达缺失与胃癌的分化程度、局部淋巴结转移和临床TNM分期有密切关系（P〈0．05，P〈0．01）。CDK4、PCNA的高表达和胃癌的淋巴结转移和临床TNM分期有密切关系（P〈0．01）。胃癌中p16、p27^kip1的表达和CDK4、PCNA的表达呈负相关（P〈0．01），而p16和p27^kip1的表达、CDK4和PCNA的表达呈正相关（P〈0．01）。结论 CDK4、PCNA的高表达和p16、p27^kip1的表达缺失与胃癌的发生以及侵袭和转移有关，可作为判断胃癌病情和预后的重要指标。     

NF-κB、cyclin D1和p27在非细胞肺癌中的表达及意义

目的：探讨核转录因子-KB（NF-κB）、细胞周期素DI（cyclin D1）和p27在非小细胞肺癌中的表达及临床意义。方法：应用免疫组化S-P法检测58例非小细胞肺癌（NSCLC）和15例正常肺组织中NF-κBp65、cyclin D1、p27的表达。结果：NSCLC组织中NF-κBp65和cyclin D1的表达明显高于正常肺组织中的表达（P〈0．01），p27表达则明显低于正常肺组织（P〈0．01）。NF-κBp65表达与NSCLC瘤体大小、淋巴结转移、临床分期密切相关（P〈0．05）cyclin D1表达与瘤体大小、肿瘤分化、淋巴结转移、临床分期密切相关（P〈0．05）；p27表达与瘤体大小、肿瘤分化密切相关（P〈0．05）。NF-κBp65与cyclin D1在NSCLC组织中的表达呈正相关（P〈0．05），cyclin D1与p27的表达呈负相关（P〈0．05），NF-κBp65与p27的表达无相关性（P〉0．05）。结论：NF-κB、cyclin D1、p27与非小细胞肺癌的发生、发展有关，检测三种蛋白的表达可以间接判断NSCLC的生物学行为。     

p57KIP2、cyclin D1和cyclin E在宫颈鳞癌中的表达及意义

目的探讨p57^KIP2、cyclin D1及cyclin E蛋白在宫颈癌发生、发展中的作用。方法用免疫组织化学SP法检测100例宫颈鳞癌、60例宫颈上皮内瘤变和30例正常宫颈鳞状上皮组织中p57^KIP2、cyclin D1和cyclin E蛋白的表达情况。结果cyclin D1、cyclin E蛋白在宫颈SCC与NE、CIN与NE组织中阳性表达率之间比较、cyclin E蛋白在宫颈SCC与CIN组织中阳性表达率之间比较，差异均有显著性（P〈0．01）；cyclin D1与cyclin E之间的表达呈正相关（P〈0．01）；三者表达均与组织学分级、淋巴结转移、患者年龄无关（P〉0．05）。结论p57^KIP2、cyclin D1及cyclin E蛋白共同参与了宫颈癌的发生发展。cyclin D1和cyclin E蛋白高表达可能是宫颈组织恶变的重要生物学标志，cyclin E异常表达是宫颈癌发生的早期事件。     

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。     

p57 KIP2 、cyclin D1 和cyclin E 在宫颈鳞癌中的表达及意义

 目的 探讨p57^KIP2、cyclin D1及cyclin E蛋白在宫颈癌发生、发展中的作用。方法 用免疫组织化学SP法检测100例宫颈鳞癌、60例宫颈上皮内瘤变和30例正常宫颈鳞状上皮组织中p57 KIP2、cyclin D1和cyclin E蛋白的表达情况。结果 cyclin D1、cyclin E蛋白在宫颈SCC与NE、CIN与NE组织中阳性表达率之间比较、cyclin E蛋白在宫颈SCC与CIN组织中阳性表达率之间比较,差异均有显著性（P〈0．01）;cyclin D1与cyclin E之间的表达呈正相关（P〈0．01）;三者表达均与组织学分级、淋巴结转移、患者年龄无关（P〉0．05）。结论 p57 ^KIP2、cyclin D1及cyclin E蛋白共同参与了宫颈癌的发生发展。cyclin D1和cyclin E蛋白高表达可能是宫颈组织恶变的重要生物学标志,cyclin E异常表达是宫颈癌发生的早期事件。     

胰腺癌组织中p57kip2、cyclinE和PCNA表达与临床病理因素的关系

背景与目的:细胞周期调控异常、细胞过度增殖与肿瘤发生密切相关 ,p57kip2蛋白作为细胞周期负性调控因子与胰腺癌的关系尚很少报道.本研究的目的是探讨 p57kip2、 cyclin E蛋白和增殖细胞核抗原( proliferating cell nuclear antigen,PCNA)在胰腺癌发生、发展中的作用.方法:应用免疫组织化学技术( SP法)对 32例胰腺癌组织及癌旁组织中 p57kip2、 cyclin E蛋白和 PCNA表达进行检测.结果: p57kip2蛋白在胰腺癌组织中的阳性表达率为 46.9% ,显著低于癌旁胰腺组织 (75.0% )(χ 2=5.317,P< 0.05),并与胰腺癌组织分化程度有关 (P< 0.05),而与淋巴结转移情况无关 (P >0.05); cyclin E蛋白阳性表达率在胰腺癌组织中为 68 8%,显著高于癌旁胰腺组织 (43.8% )(χ 2=4 063,P< 0.05) ,并与胰腺癌组织分化程度和淋巴结转移情况相关 (P< 0.05);PCNA阳性表达率在胰腺癌组织中为 71.9%,显著高于癌旁胰腺组织 (43.8% )(χ 2=5.189,P< 0.05),并与胰腺癌组织分化程度和淋巴结转移情况有关 (P< 0.05). p57kip2阳性胰腺癌组织中 cyclin E蛋白阳性表达率 ( 60.0%)低于 p57kip2阴性胰腺癌组织中 cyclin E蛋白阳性表达率( 76.5%) ,但两者无相关 (r=- 0 11211,P >0.05).结论: p57kip2蛋白低表达和 cyclin E、 PCNA蛋白过度表达与胰腺癌的发生、发展有关.     

苯并芘对HELF细胞周期及相关基因表达的影响

背景与目的： 探讨苯并芘(benzo[a]pyrene, BaP)影响人胚肺成纤维细胞(HELF)周期的途径和作用机制。 材料与方法： 用5 μmol/L的BaP处理HELF细胞后,采用细胞计数,流式细胞仪检测BaP对细胞生长和周期的影响,用PCR和Western-blotting检测BaP对细胞周期相关基因mRNA和蛋白的影响。 结果： BaP处理HELF细胞后,显著促进细胞的生长,在处理后第8 d时细胞数目约增加50％(P<0.01),细胞体积增大,促进细胞由G1期向S期和G2/M期的转换。 mRNA和蛋白检测结果显示：BaP处理细胞后可以促进p53、cyclin D1、CDK2、CDK4和p21 mRNA及蛋白的表达(P<0.05或P<0.01),抑制cyclin E mRNA及蛋白的表达(P均<0.01),而对cyclin A和p27 mRNA及蛋白表达没有明显影响(P均>0.05)。 结论： BaP对HELF细胞周期的调控逃脱了p53-p21的关卡作用,而通过诱导cyclin D1、CDK2和CDK4的表达来加快细胞周期的进程,该途径为cyclin A和p27非依赖型。     

大肠癌组织多种细胞周期蛋白表达与淋巴结癌转移数量的关系

目的 探讨大肠癌组织mdm2、p53、p21、cyclin D1、cdk4、p16蛋白表达与淋巴结癌转移及转移数量的关系。方法 应用多重Logistic回归分析筛选与淋巴结癌转移及转移数量密切相关的因素。结果 mdm2、cyclin D1是淋巴结高转移（≥3个）的促进因素,p21是淋巴结高转移的保护因素,高转移与无转移组之间mdm2、cyclin D1、p21表达的差异及高转移与低转移组之间cyclin D1、p21表达的差异有显著性,淋巴结低转移组与无转移组之间6种细胞周期蛋白表达的差异无显著性。结论 淋巴结癌转移数量与mdm2、cyclin D1过度表达及p21低表达明显相关,癌组织mdm2、cyclin D1和p21蛋白检测对常规光镜检查淋巴结未见癌转移患者的预后评估有一定意义。     

Further evidence that altered p16/CDKN2 gene expression is associated with lymph node metastasis in squamous cell carcinoma of the esophagus

Esophageal squamous cell carcinoma (ESCC) has high malignant potential with a poor outcome. Lymph node metastasis is the most useful indicator for predicting the outcome of ESCC. The p16/MTS1/CDKN2 gene and the cyclin D1/PRAD-1 gene cooperatively regulate CDK4-mediated phosphorylation of RB protein in the cell cycle. We immunohistochemically detected p16, cyclin D1, and RB expressions in both primary lesions and metastatic lymph nodes in ESCC. Among the 50 ESCC primary lesions, 24 (48%) were positive for p16, while 26 (52%) were negative for p16. Sixteen (32%) were p16-positive, 34 (68%) were p16-negative among the 50 ESCC metastatic lymph nodes. Eight cases (16%) were p16-positive in primary lesion and p16-negative in lymph node, however, no cases that was p16-negative in the primary tumor exhibited p16-positivity in metastatic lymph nodes (p < 0.0001). Seventeen (34%) of the 50 ESCC primary lesions were cyclin D1-positive, while 33 (66%) were cyclin D1-negative. Twenty-four (48%) were cyclin D1-positive, 26 (52%) were cyclin D1-negative among the 50 metastatic lymph nodes. Five cases (10%) were cyclin D1-positive in primary lesion and cyclin D1-negative in lymph node, and 12 cases (24%) were cyclin D1-negative in primary lesion and cyclin D1-positive in lymph nodes. Nine cases (18%) were RB-negative in 50 primary lesions, and the rate of loss of RB expression in metastatic lymph nodes was not markedly higher than in primary lesions. Thirty-nine (78%) of 50 primary lesions and 46 (92%) of 50 metastatic lymph nodes had altered expression of at least one of the three G1 control genes. Tumor cell with disruption of these cell cycle regulators can get a growth advantage and metastatic potential during tumor progression, especially p16/CDKN2 alterations may be associated with lymph node metastasis in ESCC. These results also suggest that tumor cells in metastatic lymph nodes may have more aggressive proliferation and higher malignant potential than tumor cells in primary lesions.     

流式细胞术测定COX-2、iNOS基因蛋白在食管癌上的表达及意义

 目的　探讨环氧化酶 2 (COX 2 ) ,Ⅱ型一氧化氮合酶 (iNOS)基因蛋白在食管癌上的表达以及与食管癌的分化和淋巴结转移的关系。方法　采用流式细胞术定量检测 6 5例食管鳞状细胞癌上COX 2、iNOS的蛋白表达量 (用荧光指数FI表示 )。结果　COX 2、iNOS在低分化型 (G3)鳞癌上的表达量明显高于分化型 (G1、G2 )鳞癌 (其P值分别为 0 .0 0 0 1、0 .0 385 ) ;二者之间呈明显正相关。这两种基因蛋白的表达强弱均与淋巴结转移无关。结论　COX 2、iNOS的表达强弱与食管癌的分化密切相关 ;COX 2与i NOS在促食管癌的发生发展中有互相协同作用     

Insufficient effect of p27(KIP1) to inhibit cyclin D1 in human esophageal cancer in vitro

The cell cycle is controlled by protein complexes composed of cyclins and cyclin-dependent kinases. p27KIP1 (p27) is one of the Kip/Cip family cyclin-dependent kinase inhibitory proteins which negatively regulate cell cycle progression, and have been proposed as candidate tumor suppressor genes. To examine the role of p27 in the development of human esophageal squamous cell carcinoma (ESCC), we performed Western blot and immunoprecipitation analyses of the levels of expression of p27 protein in a series of ESCC cell lines. This protein was expressed at various levels in these cell lines during exponential growth. p27 level was significantly associated with that of cyclin D1, but not of cyclin E. Further cell cycle synchronization studies demonstrated that p27 was free or bound with affinity to cyclin E-CDK2 more than to cyclin D1-CDK4 or cyclin D1-CDK6. It is known that overexpression of cyclin D1 rather than cyclin E is involved in the pathogenesis of ESCC. Our findings indicated that high expression of p27 throughout the G1 to S phase may inhibit more likely cyclin E, than cyclin D1, which promotes tumor growth of esophageal squamous cell carcinoma.     

非小细胞肺癌组织中PTEN/p27kip1的表达及其意义

目的检测PTEN、p27kip1、cyclin 3种蛋白在非小细胞肺癌组织的表达及与临床病理特征的关系.方法在60例非小细胞肺癌原发灶(包括37例鳞癌和23例腺癌)中应用免疫组织化学S-P法检测3种蛋白的表达.结果 60例肺癌原发灶中PTEN、p27kip1、cyclin 3种蛋白表达阳性率分别为:48.3%;41.7%;73.3%.PTEN与淋巴结转移显著相关(<0.05)p27kip1、cyclin 与淋巴结转移无关(>0.05)这3种蛋白表达均与肿瘤细胞分化程度显著相关(<0.05)与组织学类型无关(>0.05)60例非小细胞肺癌组织中,PTEN的表达与p27kip1呈显著正相关(P<0.01),而与cyclin 无明显相关(>0.05)p27kip1与cyclin 的表达之间呈显著负相关(P<0.01).结论 PTEN明显地抑制了非小细胞肺癌的浸润和转移,p27kip1表达缺失与肺癌细胞的分化有关,PTEN/p27kip1衰老诱导途径在非小细胞肺癌的恶性进展中起着很重要的作用.     

Expression of p27Kip1, cyclin E and E2F-1 in primary and metastatic tumors of gastric carcinoma.

The cell cycle is controlled by positive and negative regulators. Gene abnormalities and aberrant expressions of various cyclins/CDKs and CDK inhibitors may play a pivotal role in stomach carcinogenesis. To clarify the role of cyclin E, CDK inhibitor p27Kip1 and their target molecule, E2F-1 in tumor metastasis, we examined immunohistochemically the expression of cyclin E, p27Kip1 and E2F-1 in 23 gastric carcinomas and metastatic tumors of the lymph node. Most of gastric carcinomas with lymph node metastasis showed reduced p27Kip1 expression. p27Kip1 was negative in 39% (9/23) of primary tumors, while it was so in 52% (12/23) of lymph node metastases. By comparison of p27Kip1 expression in primary and metastatic tumors in individual cases, metastatic tumor cells in the lymph nodes were expressed at weaker levels than in those in primary tumors in 43% (10/23) of the cases. On the other hand, over 70% (17/23) and 50% (12/23) of the cases expressed cyclin E and E2F-1 at nearly the same levels in both primary tumor and lymph node metastasis, respectively. These results suggest that tumor cells with reduced p27Kip1 expression may selectively metastasize to lymph node or distant organs.     

A multi-institutional study of immunohistochemical investigation for the roles of cyclin D1 and E-cadherin in superficial squamous cell carcinoma of the esophagus

BACKGROUND AND OBJECTIVES: Aiming to clarify and possibly extend indications for minimally invasive treatment, we characterized superficial esophageal cancers (SEC) with respect to biologic properties regulating malignant potential. METHODS: Surgical specimens obtained at eight cancer institutes from 222 Japanese patients with SEC (all squamous cell carcinomas) were investigated immunohistochemically for expression of cyclin D1 and E-cadherin. RESULTS: Perturbations of cyclin D1 (overexpression) and E-cadherin (reduced expression) were observed in 37.6% (68 of 181) and 39.9% (71 of 178) of SEC patients. E-cadherin expression was more frequently reduced in cancers that invaded the submucosal layer, while cyclin D1 overexpression was constant, irrespective of depth of invasion. Overexpression of cyclin D1 was more frequent in poorly differentiated squamous cell carcinomas, while E-cadherin did not vary according to histologic differentiation. Lymph node metastasis, the only independent postoperative prognostic factor in these patients, occurred in only 4.8% of mucosal cancers (2 of 42), but in 51.1% of submucosal cancers (92 of 180). However, neither cyclin D1 nor E-cadherin status affected lymph node metastasis. CONCLUSION: Both E-cadherin and cyclin D1 play important roles in esophageal carcinogenesis, but neither can be used to identify patients who do not require lymph node dissection and might be treated by endoscopic mucosal resection.     

食管鳞状细胞癌中Wnt2、β-catenin、c-myc 和cyclinD1的表达及临床意义

 目的研究Wnt2、β-catenin及其下游的靶基因c-myc、cyclinD1在食管鳞癌中的表达情况,以期探讨其在食管鳞癌发生发展中的作用及意义。方法用免疫组织化学SP法检测40例食管鳞癌患者石蜡包埋的癌组织、癌旁组织及正常组织中Wnt2、β-catenin、c-myc、cyclinD1表达情况,分析Wnt2、β-catenin和c-myc、cyclinD1表达之间的相关性及与临床病理特征的关系。结果Wnt2、β-catenin和c-myc、cyclinD1在食管癌组织、癌旁组织及食管正常组织中表达阳性率逐渐降低,差异有统计学意义（P<0.01）,β-catenin和c-myc与肿瘤浸润和有无淋巴结转移相关（P<0.05）。结论Wnt2β-catenin Tcf4通路在食管鳞状细胞癌发生发展中起重要作用,该通路有可能成为基因治疗的潜在靶点。     

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