HCV肝炎肝硬化中核心蛋白、p14ARF和p21WAF1的表达

目的检测HCV肝炎、肝硬化组织中的核心蛋白、p14ARF、p21WAF1的表达及其相关性,探讨HCV核心蛋白对p14ARF、p21WAF1的表达作用.方法收集HCV表达阳性的肝炎、肝硬化组织23例,采用免疫组织化学EnVision两步法检测HCV感染的肝炎、肝硬化组织的核心蛋白、p14ARF、p21WAF1的表达,并分析三者间的关系.结果核心蛋白、p14ARF、p21WAF1的阳性表达主要定位于细胞核. 在核心蛋白均阳性的组织中,p14ARF的阳性表达率为69.6%,p21WAF1表达率为13%;三组间的阳性强度比较,差异有显著性(P=0.01),HCV核心蛋白与p14ARF、p21WAF1各组间的P值均<0.01;HCV核心蛋白与p14ARF、p21WAF1阳性强度两者间相关性检验,相关系数rs分别为-0.053、0.45(P值分别为0.72、0.20),表明无相关性.结论 HCV核心蛋白对p14ARF、p21WAF1的表达有影响可能间接促进p14ARF的表达,但抑制p21WAF1的表达.     

皮肤瘢痕及瘢痕癌组织中cyclin A、p21WAF1的表达及意义

目的 探讨cyclin A、p21WAF1在皮肤病理性瘢痕和瘢痕癌组织中的表达及意义.方法 以病理性皮肤瘢痕、皮肤瘢痕癌组织为研究对象,以正常皮肤组织为对照.采用免疫组化SP法分别检测cyclin A、p21WAF1蛋白的表达,采用核酸分子原位杂交法检测cyclin A mRNA、p21WAF1 mRNA的表达,结合图像分析,分别观测3组被检组织中所检各项指标的表达(平均光密度和阳性面积);所有数据输入计算机后运用SPSS 16.0软件包进行统计学分析.结果 (1)cyclin A、cyclin A mRNA在正常皮肤表皮和病理性皮肤瘢痕上皮中的表达呈阴性或弱阳性,在瘢痕癌组织中呈强阳性.瘢痕癌组的表达(平均光密度和阳性面积)与正常皮肤组及皮肤瘢痕组比较,差异均有统计学意义(P＜0.01),但正常皮肤组与瘢痕组比较,差异无统计学意义(P＞0.05).(2)p21WAF1蛋白、p21WAF1 mRNA在正常皮肤表皮和病理性皮肤瘢痕上皮中的表达呈阴性或弱阳性,在瘢痕癌组织中呈强阳性.瘢痕癌组的表达(平均光密度和阳性面积)与正常皮肤组及皮肤瘢痕组比较,差异均有统计学意义(P<0.01),但正常皮肤组与瘢痕组比较,差异无统计学意义(P>0.05).(3)相关分析显示,在皮肤瘢痕癌中,cyclin A 与cyclin A mRNA(r=0.766,P<0.01)、p21WAF1与p21WAF1 mRNA(r=0.791,P<0.01)的表达呈正相关;cyclin A与p21WAF1蛋白(r=0.656,P<0.01)的表达呈正相关;cyclin A mRNA与p21WAF1 mRNA的表达呈正相关(r=0.475,P<0.05).结论 (1)cyclin A及其mRNA的高表达,可能与皮肤瘢痕癌的发生有关;(2)在瘢痕癌中,cyclin A、p21WAF1同时存在蛋白水平和基因水平的异常表达;(3)p21WAF1及其mRNA在瘢痕癌中的高表达可能与细胞周期调控的反馈机制有关.     

宫颈鳞癌中p16、p21WAF1、Rb、cyclinE蛋白的表达

目的通过观察p16、p21WAF1、Rb、cyclinE 4种细胞周期相关蛋白在宫颈鳞状细胞癌中的表达,探讨它们在宫颈癌的细胞周期调控中的作用.方法采用免疫组化Eli Vision二步法对88例宫颈鳞癌组织,16例宫颈上皮内病变(CIN)组织,15例宫颈炎组织进行p16、p21WAF1、Rb、cyclinE 4种蛋白表达的检测.结果p16、p21WAA1、cyclinE在宫颈癌中的表达高于宫颈炎(P＜0.05);Rb在宫颈癌的表达少于宫颈炎(P＜0.05);Rb与p16在宫颈癌中的表达呈负相关(r=-0.675,P＜0.05).结论宫颈癌细胞周期G1期中,由于Rb蛋白的缺失,使cyclinE表达升高,致使癌细胞增生;同时,p16、p21WAF1蛋白在宫颈癌中的表达增高,并失去抑制作用.     

胰腺癌p53上下游基因mdm2、p21WAF/CIP1以及P14ARF蛋白的表达及相互关系

目的 探讨p14ARF、p53、mdm2及p21WAF/CIP1蛋白在胰腺癌组织中的表达、相互关系及意义。方法 选取167例胰腺癌、101例癌旁与13例良性病变组织构建组织芯片(又称组织微阵列),应用免疫组织化学EnVision二步法检测这4种蛋白在胰腺良恶性病变中的表达。结果 p14ARF、p53、mdm12及p21WAF/CIP1在胰腺癌中表达的阳性率分别为35.3％(59/167)、57.5％(96/167)、64.1％(107/167)和39.5％(66/167)。同癌旁组织相比,p53和mdm2表达明显升高(P<0.01),而p14ARF和p21WAF/CIP1的表达明显降低(P<0.05)。p21WAF/CIP1的阳性表达与年龄、神经受累显著相关(P<0.05);p53的阳性表达与肿瘤的分化、淋巴结转移和神经受累均显著相关(P<0.05);mdm2的阳性表达与肿瘤的分化显著相关(P<0.05);p14ARF与年龄和浸润转移显著相关(P<0.05)。四者阳性表达两两之间统计学上具有关联性(P<0.05)。结论 p53和mdm2的过表达以及p14ARF和p21WAF/CIP1的缺失表达可能会导致胰腺癌的形成和进展;4种蛋白主要以p14ARF-p53-mdm2-p21WAF/CIP1通路的方式作用于细胞的转化和肿瘤的形成;联合检测p53和mdm2的表达可用于评定胰腺癌的恶性程度。     

p53、p21~(WAF1)蛋白在非小细胞肺癌中的表达及其临床意义

目的　探讨原发性非小细胞肺癌中p5 3、p2 1WAF1蛋白表达与临床病理及预后的关系。方法　应用免疫组织化学 (SP法 )方法。共检测非小细胞肺癌 147例 ,其中腺癌 6 6例 ,鳞癌 6 3例 ,腺鳞癌 14例 ,大细胞癌 4例。结果　p5 3蛋白总阳性率为 6 1.2 % (90 / 147) ,腺癌为 5 7.6 % (38/ 6 6 ) ,鳞癌阳性率为 6 3.5 % (4 0 / 6 3) ,腺鳞癌为 71.4% (10 / 14) ,大细胞癌 2例阳性。p2 1WAF1蛋白总阳性率为40 1% (5 9/ 147) ,腺癌为 42 .4% (2 8/ 6 6 ) ,鳞癌为 41.3% (2 6 / 6 3) ,腺鳞癌 2 8.6 % (4 / 14) ,大细胞癌 1例阳性。肺腺癌p5 3蛋白阳性表达与其预后相关 ,6 6例腺癌中 ,生存率低于 3年组和高于 3年组的p5 3蛋白阳性率分别为 75 % (2 1/ 2 8)和 44 .7% (17/ 38) ,差异有显著性意义 (P <0 .0 2 5 )。p2 1WAF1阳性表达与肺癌预后有关 ,p2 1WAF1阳性表达者 3年生存率 (6 4.4% )高于阴性表达者 (4 6 .6 % ) (P <0 .0 5 )。p5 3阳性而p2 1WAF1阴性的非小细胞肺癌患者的预后比p5 3阴性而p2 1WAF1阳性者差 (P <0 .0 1)。结论　检测p5 3蛋白表达可作为判断肺腺癌预后的指标之一 ;检测p2 1WAF1蛋白表达有利于对非小细胞肺癌预后的判断 ;联合检测p5 3、p2 1WAF1蛋白对判断非小细胞肺癌的预后有重要的意义 ,似可作     

胃癌及其癌前病变中细胞凋亡及其调控基因的表达

目的:通过观察胃癌及其癌前病变中细胞凋亡及其调控基因p53、bcl-2、c-myc的表达,探讨其在胃癌恶性转化进程中的作用。方法:利用DNA末端标记技术(TdT-mediateddUTP-biotinnickendlabeling,TUNEL法)和免疫组织化学(streptavidin/peroxidase,S-P法),原位观察了46例胃癌、20例癌前病变、24例正常粘膜中的凋亡细胞和p53、bcl-2、c-myc的表达。结果:癌前病变和胃癌中凋亡指数显著高于正常粘膜(P＜0.01),癌前病变组高于胃癌组(P＜0.01)。p53、bcl-2、c-myc蛋白在胃癌中阳性率分别为60.9%、67.4%、73.2%,均高于正常粘膜组(P＜0.01);在癌前病变中3种蛋白阳性表达率分别为40%、95%、58.8%(P＜0.01)。胃癌中p53、bcl-2蛋白阳性细胞凋亡指数分别低于阴性组(P＜0.05),c-myc蛋白阳性组细胞凋亡指数高于阴性组(P＜0.01)。结论:细胞凋亡调控异常在胃癌发生中可能起重要作用。p53、bcl-2蛋白分别抑制细胞凋亡,c-myc蛋白在胃癌中促进细胞凋亡。     

人脑神经胶质瘤中p16、p21~(WAF1/CIP1)蛋白表达的研究

目的　探讨 p16、p2 1WAF1/CIP1两种抑癌基因与人脑神经胶质瘤恶性程度的关系。方法　采用SABC免疫组织化学方法对 6 4例人脑胶质瘤组织及 8例正常脑组织标本中p16和 p2 1WAF1/CIP1表达情况进行检测 ,并进行相关分析。结果　①p16和 p2 1WAF1/CIP1阳性表达率在人脑胶质瘤中分别为 4 5 .3%和 6 4 .1%与正常脑组织中的表达情况差异显著 (P <0 0 5 ) ;②p16蛋白和 p2 1WAF1/CIP1蛋白阳性表达率均随着胶质瘤的恶性程度的增高而降低 ,差异显著 (P <0 0 1) ,且呈负相关关系 ;③p16蛋白和 p2 1WAF1/CIP1蛋白可协同表达 ,且在正常脑组织和脑胶质瘤各分级中 p16蛋白和 p2 1WAF1/CIP1蛋白协同表达率差异显著 (P <0 0 5 ) ,并呈负相关关系。结论　p16与p2 1WAF1/CIP1蛋白的阳性表达率及协同表达率可在一定程度上反映胶质瘤细胞的恶性程度 ,可作为判断其恶性程度的有效指标     

星形细胞瘤中PTEN、Mdm2和p53的表达及其相关性研究

目的: 探讨不同组织病理分级的星形细胞瘤中PTEN、Mdm2和p53的表达水平, 并分析PTEN影响Mdm2、p53表达的信号转导机制。方法: 采用免疫组织化学方法检测68例星形细胞瘤标本中PTEN、Mdm2和p53的表达水平。结果:星形细胞瘤中PTEN、Mdm2、p53的表达水平分别是54.4%(37/68)、41.2%(28/68)、45.6%(31/68)。PTEN阳性标本中Mdm2的表达率(24.3%, 9/37)与PTEN阴性标本中该蛋白的表达率(61.3%, 19/31)相比差异显著, 统计学分析显示PTEN表达与Mdm2表达呈负相关(P＜0.01)。Mdm2表达和p53表达一致符合率达66.2%(45/68), 两者的表达密切相关(P＜0.05)。结论: (1)PTEN、Mdm2和p53表达与星形细胞瘤的组织病理分级相关。(2)抑癌基因PTEN可以下调癌基因Mdm2的表达水平。(3)Mdm2和p53的表达存在一致性。     

p21WAF1基因表达对人骨肉瘤预后的价值

目的 探讨骨肉瘤组织中p21^WAF1基因的表达与骨肉瘤生物学特性及预后的关系。方法 采用原位杂交及免疫组织化学（LSAB法）检测p21^WAF1mRNA及p21蛋白在45例骨肉瘤、10例骨纤维结构不良实体瘤组织标本的表达。结果 （1）p21蛋白阳性表达率在骨肉瘤中为17．7％（8／45）;（2）骨肉瘤组织高分化组与低分化组的p21蛋白阳性表达率之间的差异有统计学意义（40．0％,11．4％,X^2=4．34,P〈0．05）;（3）p21^WAF1mRNA表达阳性率在骨肉瘤中为42．2％（19／45）,骨肉瘤组织高分化组与低分化组的p21^WAF1mRNA阳性表达率之间的差异有统计学意义（60．0％,37．1％,X^2=20．6,P〈0．01）;（4）p21^WAF1mRNA表达阳性者术后生存时间高于表达阴性者术后生存时间（P〈0．05）。结论 （1）随着骨肉瘤恶性度的升高,p21^WAF1基因mRNA及p21蛋白的表达下降。（2）p21WAF1基因mRNA在骨肉瘤中的表达对评价患者的预后有一定价值。     

p57和p53蛋白在水肿性流产及葡萄胎鉴别诊断中的作用

目的探讨p57和p53蛋白在水肿性流产、完全性和部分性葡萄胎鉴别诊断中的作用。方法分别收集正常绒毛、水肿性流产、部分性葡萄胎和完全性葡萄胎石蜡标本10、12、23和20例,应用免疫组织化学EnVision法检测p57和p53蛋白在这些组织中的分布及表达水平。结果p57蛋白在正常绒毛、水肿性流产及部分性葡萄胎组织中主要分布于绒毛的细胞滋养细胞及间质细胞,阳性表达比例分别为10/10、12/12和100％( 23/23),各组间相比差异无统计学意义(P＞0.05)。在完全性葡萄胎中细胞滋养细胞及间质细胞p57表达缺失,与部分性葡萄胎相比,差异有统计学意义(P＜0.05)。p53蛋白主要表达于完全性和部分性葡萄胎的细胞滋养细胞及中间滋养细胞。在正常绒毛中p53蛋白呈阴性表达,水肿性流产中仅1例p53蛋白呈阳性表达(1/12),部分性葡萄胎和完全性葡萄胎中p53蛋白的阳性率分别为60.9％( 14/23)和85.0％ (17/20);p53蛋白的阳性率,部分性葡萄胎较水肿性流产明显增加,完全性葡萄胎较部分性葡萄胎也明显增加,差异均有统计学意义(均P ＜0.05)。结论p57蛋白免疫组织化学检测可辅助鉴别完全性和部分性葡萄胎,而p53蛋白的检测则有助于鉴别水肿性流产和部分性葡萄胎。     

Molecular analysis of p53, MDM2 and H-ras genes in low-grade central osteosarcoma

Low-grade central osteosarcoma is an uncommon form that is characterized by a long premorbid history, and is compatible with prolonged survival after treatment. However, molecular abnormalities are rare in low-grade central osteosarcomas, whereas p53 mutations occur in approximately 20% of conventional high-grade osteosarcomas. In this study, 21 cases of low-grade central osteosarcoma were analyzed for mutations of the p53 gene, amplification of the MDM2 gene, and mutations of the H-ras gene using formalin-fixed, paraffin-embedded materials. We also examined the expression of p53, MDM2, and p21WAF1 protein immunohistochemically and assessed the proliferation activities using the monoclonal antibody MIB-1. One case (4.7%) showed strong p53 immunoreactivity, whereas p53 gene mutations were not detected at all. Seven cases (33.3%) showed immunoreactivity for MDM2 protein. As for gene alterations, MDM2 amplification was found in four cases (19.0%). p21WAF1 expression was detected in 12 cases (57.1%). MIB-1-LI showed very low levels in all the cases and no significant correlation with p53 or MDM2 immuno-reactivity. None of the tumors showed H-ras mutations. In conclusion, the number of p53 gene alterations in low-grade central osteosarcomas is lower than that in conventional high-grade osteosarcomas. MDM2 alterations and p21WAF1 expression might be involved in the tumorigenesis of low-grade central osteosarcomas.     

p53 expression in CMV-infected cells: association with the alternative expression of the p53 transactivated genes p21/WAF1 and MDM2

The p53 tumour suppressor gene is a cell cycle regulator, able to induce cell cycle arrest to allow DNA repair or apoptosis. The molecular mechanisms underlying p53 action imply transactivation of p53 dependent genes such as WAF1 (for wild type p53 associated fragment 1) and the murine double minute (MDM2) gene. In some cases, inactivation of the p53 gene results from p53 gene mutations leading to p53 protein accumulation, but in others it may results from mechanisms other than mutation, such as interaction with viral or cellular proteins. The expression of p53 protein and p53 transactivated gene proteins p21/WAF1 and MDM2, combined with in situ detection of apoptosis, was studied in specimens of CMV-infected patients as an in vivo model of p53 alteration not due to point mutation. p53 positivity was found in CMV + cells in different tissues, in cells with typical inclusion bodies, and in in situ hybridization and immunohistochemistry CMV + cells without inclusions (hidden infection). Although this p53 reactivity was accompanied by the expression of MDM2 and p21/WAF1 proteins, the patterns of MDM2 and p21/WAF1 protein expression were mutually exclusive, and were associated with the presence or absence of inclusion bodies. Nuclei bearing inclusion bodies were usually MDM2 +, p21/WAF1?, while hidden infected cells were usually MDM2?, p21/WAF1 +. Apoptosis was not detected in any tissue section from CMV-infected patients. Two alternative patterns were found in CMV-infected tissues: p53 +, p21/WAF1 +, MDM2?, or p53 +, p21/WAF1?, MDM2 + protein expression. These may represent examples of p53 dependent alternative effects in the course of CMV infection. Early stages are represented by CMV + cells without inclusion bodies, which display p53 and p21/WAF1 expression, suggesting that p53 could be acting as a growth suppressor protein. Late CMV infection is represented by cells harbouring inclusion bodies. These cells showed a p53 +, p21/WAF1?, MDM2 + profile, consistent with MDM2 mediated p53 inactivation. The absence of p21/WAF1 expression and lack of apoptosis suggest that the p53 protein expressed by MDM2 + cells could be functionally inactivated in CMV-infected cells with inclusion bodies. Previous studies have suggested that p53 inactivation by MDM2 over-expression occurs in sarcomas and lymphomas. Our observations seem to indicate that this mechanism of MDM2 mediated p53 inactivation may play a role in the late phase of CMV infection.     

nm23—H1与p21、p53蛋白在人卵巢癌中表达的相关性研究

目的：研究人卵巢癌组织中肿瘤转移抑制基因nm23-H1、抑癌基因p21^WAFI及p53蛋白的表达,探讨它们在卵巢癌发生、发展中的作用。方法：采用免疫印迹技术,对74例卵巢癌组织及21例卵巢非癌组织中nm23-H1p21^WAFI及p53蛋白进行检测。结果：在卵巢癌中nm23-H1、p53蛋白的表达高于卵巢非癌组织,p21^WAFI蛋白的表达呈下调;肿瘤发生转移时卵巢癌组织中nm23-H1和p21WAF蛋白表达显著低于未发生肿瘤转移的癌组织。p53蛋白表达与肿瘤转移无关。nm23-H1 \p21^WAFI 蛋白与卵巢癌组织类型无关,但与卵巢癌的临床分期相关。p53蛋白的表达与卵巢癌组织类型及卵巢癌的临床分期无关。nm23-H1与p53、p21^WAFI蛋白间表达在卵巢癌中呈相关性。结论：nm23-H1、p53、p21^WAFI 基因在卵巢癌发生、发展中起一定的作用,nm23-H1与p53、p21^WAFI基因在卵巢癌发生、发展中可能起协同作用。     

p21WAF1/CIP1 expression in colorectal carcinoma correlates with advanced disease stage and p53 mutations

Defects in the mechanisms controlling the cell cycle are crucial in cell transformation and/or tumour progression. p21^WAF1/CIP1 is an inhibitor of cyclin-dependent kinases, induced by p53-dependent and p53-independent pathways, which can block progression through the cell cycle. p21^WAF1/CIP1 expression has been investigated immunohistochemically in a series of 191 patients with colorectal cancer of known p53 status. The purpose of the study was two-fold: to assess the relationship between p21^WAF1/CIP1 immunoreactivity and p53 alterations, and to evaluate the prognostic significance of p21^WAF1/CIP1 expression. In 96 carcinomas (51 per cent), p21^WAF1/CIP1 was expressed in over 10 per cent of tumour cells, whereas in 26, p21^WAF1/CIP1 was detected in under 10 per cent of neoplastic cells; 69 tumours lacked p21^WAF1/CIP1 expression. Immunoreactivity was more frequent in tumours of the right colon (p < 0·003) and was inversely correlated with tumour stage (p < 0·03), p53 gene mutations (p < 0·0007), p53 protein accumulation (p < 0·019), and Bcl-2 expression (p < 0·0005). In univariate analysis, down-regulation of p21^WAF1/CIP1 expression was associated with poor overall (p = 0·0022) and disease-free survival (p = 0·0009). Multivariate analysis, however, did not confirm any independent prognostic significance of p21^WAF1/CIP1 expression. The results indicate that p21^WAF1/CIP1 is associated with abnormal accumulation of p53 protein and the occurrence of p53 gene mutations in colorectal cancer and that lack of p21^WAF1/CIP1 expression is correlated with reduced patient survival in univariate analysis. These data underline the crucial pathogenetic role of the p53–p21^WAF1/CIP1 pathway in carcinomas of the large bowel. Copyright © 1999 John Wiley & Sons, Ltd.     

Relationship between abnormal p53 protein and failure to express p21 protein in human breast carcinomas

Alterations in the p53 protein are a common feature in most malignancies, including breast carcinomas. p53 protein alterations contribute to malignant transformation in several ways, through genomic instability and accumulation of additional genetic alterations in other genes, through alteration of the p53-dependent apoptotic pathway, and through downregulation of downstream effector proteins such as p21 (WAF1/CIP1), necessary for cell-cycle growth arrest. Cell-cycle arrest is needed to allow DNA repair after injury. This study examines the relationship between abnormalities in p53 protein and expression of p21 protein in 70 cases selected from a series of 212 sporadic human breast carcinomas. Immunohistochemistry (IHC) was used for detection of p53 and p21 protein expression. Constant denaturant gel electrophoresis (CDGE) was used for detection of mutations in exons 5–8 of the TP53 gene. A highly significant association was found between abnormalities in p53, scored as protein accumulation and/or mutations, and lack of p21 expression. p21 was also shown to be downregulated in samples without p53 alterations, indicating that other mechanisms are also involved in turning off this gene. © 1997 John Wiley & Sons, Ltd.