骨肉瘤p21~(ras)蛋白、p21~(waf1/cip1)蛋白、PCNA与AgNOR的表达及其临床意义

目的探讨骨肉瘤组织中p21ras蛋白、p21waf1/cip1蛋白、PCNA与AgNOR的表达及其临床意义。方法应用免疫组化方法和组织化学方法检测45例骨肉瘤病人的骨肉瘤组织、20例骨软骨瘤病人的骨软骨瘤组织和20例正常骨组织中p21ras蛋白、p21waf1/cip1蛋白、PCNA与AgNOR的表达情况,对有关临床病理指标综合分析。结果45例骨肉瘤组织中p21ras蛋白、p21waf1/cip1蛋白和PCNA的阳性表达率分别为86.7%、13.3%和100%,AgNOR计数为9.63±2.47,阳性表达率与AgNOR计数均高于骨软骨瘤和正常骨组织;p21ras蛋白的表达与PCNA的表达存在平行关系;AgNOR计数与PCNA的表达呈正相关;四者的表达与骨肉瘤有关临床病理指标之间存在一定的关系。结论检测p21ras蛋白、p21waf1/cip1蛋白、PCNA与AgNOR为骨肉瘤恶性程度和预后的评估提供了重要的辅助参考指标。     

骨肉瘤组织SRp20蛋白表达及其临床意义的初步研究

目的:探讨骨肉瘤组组织SRp20蛋白表达情况及其临床意义.方法:应用Max Vision免疫组化方法检测64例骨肉瘤组织和18例骨软骨瘤组织SRp20蛋白表达情况,并分析与临床病理参数之间的关系.结果:骨肉瘤组织和骨软骨瘤组织中,SRp20胞核阳性表达率分别为14.1％(9/64)和0(0/18),P=0.195;而骨肉瘤组织和骨软骨瘤组织中,SRp20细胞质阳性表达率分别为73.4％(47/64)和5.6％(1/18),P＜0.001.在骨肉瘤组织中,SRp20的细胞质阳性表达与骨肉瘤的恶性程度(P=0.002)、Enneking分期(P=0.001)以及5年生存率(P=0.023)相关.结论:SRp20细胞质表达与骨肉瘤患者的不良预后相关,可能成为预测患者预后情况的靶点.     

骨肉瘤P21^WAF1蛋白、PCNA与Ki—67的表达及其意义

目的：研究骨肉瘤组织中P21^WAF1蛋白、增殖细胞核抗原（PCNA）、Ki-67蛋白的表达，及其与骨肉瘤的发生、发展之间的关系及对预后的影响。方法：用免疫组化LSAB法检测P21^WAF1蛋白、增殖细胞核抗原（PCNA）、Ki-67蛋白在骨肉瘤中的表达。结果：45例骨肉瘤中，P21^WAF1有8例阳性表阳，阳性率为17.77%；PCNA均有表达，阳性率为100%；Ki-67有27例阳性表达，阳性率为60%。结论：P21^WAF1的低表达、PCNA、Ki-67的高表达，从一个侧面反映了骨肉瘤的恶性程度较高，检测P21^WAF1蛋白、增殖细胞核抗原（PCNA)、Ki-67蛋白等多个指标可较全面反映骨肉瘤的恶性程度的高低及患者的预后情况。     

p27蛋白与PCNA在胃癌中的表达及其临床意义

目的 探讨p27及PCNA在胃癌中的表达意义及相互关系。方法 应用免疫组化S—P法对62例胃癌进行p27蛋白和PCNA的检测。结果 胃癌组织中p27、PCNA表达的阳性率分别为41．9％、83．9％．p27的表达与Bomman分型、肿瘤浸润深度、淋巴结转移、临床分期有关；PCNA仅与肿瘤分化程度有关，p27与PCNA表达呈负相关。结论 胃癌组织中p27、PCNA的表达可能是反映胃癌生物学行为和预测预后的重要指标。     

p16、p27蛋白在骨巨细胞瘤和骨肉瘤中的表达

目的：探讨抑癌基因p16、p27在骨巨细胞瘤（GCT）和骨肉瘤（OS）中的蛋白表达特点及其在骨肿瘤发生中的意义。方法：应用SABC免疫组织化学方法检测p16、p27蛋白在16例骨肉瘤、46例骨巨细胞瘤（其中JaffeⅠ级18例、Ⅱ级6例、Ⅲ级12例）。中的表达。结果：p16和p27蛋白的阳性表达均定位于骨肉瘤细胞、骨巨细胞瘤的基质细胞的细胞核和细胞质内。在骨巨细胞瘤Ⅰ级、Ⅱ级、Ⅲ级和骨肉瘤中p16蛋白的阳性表达率依次为16．7％、62．5％、66．7％和25％;p27蛋白的阳性表达率依次为11．8％、56．3％、50％和25％。p26和p27蛋白的骨巨细胞瘤I级中的阳性表达率显著地低于Ⅱ级和Ⅲ级（P＜0．05）,p16蛋白在骨肉瘤与骨巨细胞瘤Ⅲ级之间的阳性率差异有显著性意义（P＜0．05）,而p27蛋白在骨肉瘤和骨巨细胞Ⅲ级之间的率差异则不具有显著性意义（P＞0．05）。结论：p16、p17基因在骨巨细胞瘤和骨肉瘤的发生中起重要作用,但其阳性表达率不足以单独作为判断恶性程度和预后的指标。     

骨肉瘤P21~(WAF1)蛋白、PCNA与Ki-67的表达及其意义

目的 :研究骨肉瘤组织中P2 1WAF1 蛋白、增殖细胞核抗原 (PCNA)、Ki 6 7蛋白的表达 ,及其与骨肉瘤的发生、发展之间的关系及对预后的影响。方法 :用免疫组化LSAB法检测P2 1WAF1 蛋白、增殖细胞核抗原 (PCNA)、Ki 6 7蛋白在骨肉瘤中的表达。结果 :45例骨肉瘤中 ,P2 1WAF1 有 8例阳性表达 ,阳性率为 17.77% ;PCNA均有表达 ,阳性率为 10 0 % ;Ki 6 7有 2 7例阳性表达 ,阳性率为 6 0 %。结论 :P2 1WAF1 的低表达、PCNA、Ki 6 7的高表达 ,从一个侧面反映了骨肉瘤的恶性程度较高 ,检测P2 1WAF1 蛋白、增殖细胞核抗原 (PCNA)、Ki 6 7蛋白等多个指标可较全面反映骨肉瘤的恶性程度的高低及患者的预后情况。     

骨肉瘤中p16和生存素蛋白的表达及其相关性研究

目的探讨凋亡抑制蛋白生存素、抑癌基因p16在骨肉瘤中的表达与骨肉瘤的生物学特性的关系。方法采用免疫组化SP法,检测52例骨肉瘤、15例骨软骨瘤及15例正常骨质标本的生存素蛋白和p16蛋白表达。结果生存素蛋白在骨肉瘤中的表达显著高于骨软骨瘤和正常骨质（P〈0.05）,在肿瘤的侵袭转移方面差异有显著性（P〈0.05）,而在性别、年龄、肿瘤直径、病理类型、病理分级及临床分期等方面差异无显著性（P〉0.05）。p16蛋白在骨肉瘤中的表达显著低于骨软骨瘤和正常骨质（P〈0.05）,在肿瘤的病理分级方面差异有显著性（P〈0.05）,而在性别、年龄、肿瘤直径、病理类型、侵袭转移及临床分期等方面差异无显著性（P〉0.05）。在骨肉瘤p16蛋白表达阳性、阴性组中生存素蛋白表达的阳性率分别为73.9%和66.7%（χ^2=0.142,P=0.707）,经统计学分析,生存素蛋白和p16蛋白表达之间无明显相关（r=0.094,P=0.509）。结论凋亡抑制蛋白生存素在侵袭转移的骨肉瘤组呈高表达;抑癌基因p16在骨肉瘤中随着病理分级的降低,阳性表达率有降低趋势。提示凋亡抑制蛋白生存素、抑癌基因p16在骨肉瘤的发生和发展的不同环节中起重要作用,测定生存素蛋白表达可作为预测预后的重要参考指标,测定p16蛋白表达可作为判断骨肉瘤恶性程度的重要参考指标,并将为肿瘤的基因治疗提供新的靶点。     

PBF在骨肉瘤中的表达及其临床意义

目的:探讨PBF(papillomavirus binding factor)在骨肉瘤组织中的表达与临床病理特征之间的关系。方法:应用免疫组织化学方法检测70例骨肉瘤组织、30例骨软骨瘤及10例正常组织中PBF的表达。结果:骨肉瘤标本中PBF表达的阳性率是89%(62/70),而在骨软骨瘤标本中表达的阳性率是10%(3/30),两者比较P<0.01。PBF蛋白表达与患者年龄、性别、肿瘤体积及病理分型无关(P>0.05),而与临床Ennecking分期、有无转移有关(P<0.05)。结论:PBF的表达可能在骨肉瘤的发生发展及转移过程中起到重要作用,并可能作为骨肉瘤诊断的特异性指标。     

PTEN和p53在骨肉瘤中的表达及其临床意义

目的研究骨肉瘤组织中PTEN,p53的表达及其相关性,探讨其在骨肉瘤发生发展中的作用。方法用S-P免疫组化方法检测PTEN和P53在40例骨肉瘤及20例骨软骨瘤中的表达。结果PTEN在骨良性肿瘤组织中阳性表达率为90.0%,显著高于骨肉瘤组织的55.0%(P<0.05);p53在骨良性肿瘤组织中不表达,在骨肉瘤组织中阳性表达率为57.5%,差异有显著性(P<0.05);骨肉瘤组织中,PTEN和p53的表达呈负相关(r=-0.3710,P=0.0189)。PTEN的表达与骨肉瘤分化程度有关,与肺转移无关。p53的表达与骨肉瘤分化程度及肺转移无关。结论PTEN和p53与骨肉瘤的发生有关,且两种指标具有相关性,临床上联合检测PTEN和p53的表达对骨肉瘤的早期诊断具有指导意义。PTEN检测对判断骨肉瘤的预后具有指导意义。     

p21活化激酶5在骨肉瘤组织中的表达及临床意义

背景与目的：p21活化激酶5(p21-activated kinase 5,PAK5)为PAKs家族中新近发现的成员,在细胞骨架重组,细胞生长、增殖、分化,基因转录及细胞凋亡等一系列的细胞功能中发挥重要作用。近来有研究发现PAK5参与了胃癌、肠癌等恶性肿瘤的发生发展过程。本研究旨在探讨PAK5在人骨肉瘤组织中的表达及其与骨肉瘤患者预后的关系。方法：应用免疫组化方法检测92例骨肉瘤活检样本石蜡组织中PAK5蛋白的表达,33例骨巨细胞瘤样本作为阴性对照,并分析PAK5表达与骨肉瘤患者临床参数及生存之间的关系。结果：骨肉瘤组织中PAK5蛋白表达阳性率为71.7%(66/92),骨巨细胞瘤中未见PAK5表达(0/33)。PAK5表达与患者的性别、年龄、肿瘤部位、肿瘤大小、组织学类型、局部复发等无关,而与Enneking分期、肿瘤坏死率和肺转移相关,且PAK5高表达能降低化疗的有效率。单因素分析表明,肿瘤大小、Enneking分期、局部复发、肺转移及PAK5表达水平是影响骨肉瘤患者预后的因素。多因素分析显示,PAK5表达水平(P=0.001)及肺转移(P=0.015)是影响骨肉瘤预后相关的独立的重要因素。结论：PAK5表达水平与Enneking分期、肿瘤坏死率和肺转移相关;PAK5的表达可作为骨肉瘤预后预测的指标;PAK5高表达能降低化疗的有效率。     

Early g1 induction of p21/waf1/cip1 in synchronized osteosarcoma cells is independent of p53

Enzymatic activities of cyclin-dependent protein kinases (cdks) are tightly regulated at the level of subunit composition, involving both positive (cyclins) and negative (p21Wafl/Cipl); p16Ink4; p27Kipl) effectors. In the present study, we examined the expression of p21/WAF1/CIP1 in highly synchronized human MG-63 osteosarcoma cells in which the sequential induction of specific cyclins was characterized previously (1). Two distinct peaks of p21/WAF1/CIP1 expression were detected by Northern analysis of serum-stimulated cells: one peak was detectable by 1 hour, reached a maximum at approximately 3 hours, and diminished markedly by 4-6 hours; and a second peak was observed during S phase. The early G1 induction of the 21 kDa gene product was further demonstrated by Western blotting. Both Northern analysis and Western blotting for the p53 tumor suppressor confirmed previous reports that its expression is not detectable in MG-63 cells at any time. The transient induction of p21/WAF1/CIP1 in early G1 was correlated with a transient decrease in p9Ckshs1-agarose precipitable histone H1 kinase activity, as determined by in vitro kinase assays. In contrast, the myelin basic protein kinase activity of anti-Cdk4 immune complexes was not attenuated to a significant extent. Taken together, these studies identify a novel biochemical pathway of p21/WAF1/CIP1 induction operating in p53-deficient osteosarcoma cells; a pathway whose independence from p53 may conceivably be exploited to therapeutic advantage in the treatment of proliferative disorders.     

Prognostic impact of p21/waf1/cip1 in colorectal cancer

In addition to the tumor suppressor gene p53, Cyclin Dependent Kinases (CDK) are well known to influence the cell cycle in normal human tissues and various neoplasias as well. The purpose of our present study was to evaluate the expression of the CDK-inhibitor p21/waf1/cip1 in colorectal cancer with special emphasis on the prognostic impact. Between 1985 and 1991, 294 patients (median age, 65 years) underwent surgical operative therapy for colorectal cancer. Formalin-fixed and paraffin-embedded tumor specimens were investigated. For immunohistochemistry the Catalysed Reporter Deposition (CARD) technique was performed. The survival probability was calculated and possible prognostic risk factors were tested using multivariate analysis. The p21/ waf1/cip1 staining pattern was positive in 197 (67%) specimens and negative in 97 (33%) samples. No significant correlation could been calculated between p21/waf1/cip1 expression and other variables such as age, sex, WHO-Classification, localisation, grading, TNM-classification or UICC-stage. Patients with a positive staining reaction had a significantly better survival (p < 0.0052). Moreover, p21/waf1/cip1 was shown to be an independent prognostic parameter by multivariate analysis (p < 0.022). In contrast with these findings, the p53 tumor status had no impact on survival. P21/ waf1/cip1 appears to be an independent prognostic parameter in colorectal cancer and is associated with a favorable survival. This feature may be related to a cell cycle arrest in the G1 phase induced by p21/waf1/cip1, resulting in lower tumor cell proliferative activity.     

Expression of p53 and p21waf1/cip1 in gastric carcinoma: lack of inter-relationship or correlation with prognosis.

AIMS: The cell cycle regulators p53 and p21waf1/cip1 are expressed variably in human cancers. We investigated their expression in gastric carcinoma and determined their inter-relationship and prognostic significance. METHODS: Immunohistochemistry was used to determine their expression in material from 100 resected specimens of gastric carcinoma, and comparison was then made of the degree of expression between each, with conventional clinicopathological indices and with survival. RESULTS: Positivity was found with p53 (40%) and p21 (75%). There was no significant correlation between the expression of each individual marker, nor between each marker and 5-year survival. There appeared to be an association between p53 expression and lymph node metastases, and a higher frequency of p21waf1/cip1 expression in males. CONCLUSIONS: The expression of p53 and p21waf1/cip1 as detected by immunohistochemistry were of no value in predicting the prognosis of patients with gastric carcinoma.     

FHIT、p21waf1/cip1基因在膀胱移行细胞癌中的表达与意义

目的:通过测定脆性组氨酸三联体基因(FHIT)及p21waf1/cip1基因在膀胱移行细胞癌组织、正常膀胱组织中的表达,探讨FHIT基因以及p21waf1/cip1基因与膀胱癌的关系及其临床意义。方法:采用免疫组织化学SP法对43例膀胱移行细胞癌(BTCC)组织及14例正常膀胱组织中的FHIT基因及p21waf1/cip1基因的蛋白表达进行检测。结果:FHIT蛋白表达与肿瘤的分期、分级无相关性(P>0.05)而p21waf1/cip1蛋白的表达与之有相关性(P<0.05);FHIT蛋白的表达在GI肿瘤、浅表性肿瘤中明显低于在正常膀胱组织中的表达(P<0.05)而p21waf1/cip1蛋白的表达在上述组织比较中无差别(P>0.05);FHIT蛋白的表达在初发肿瘤中与复发肿瘤中无明显差别(P>0.05)而p21waf1/cip1蛋白的表达在上述两种组织比较有明显差别(P<0.05)。FHIT蛋白与p21waf1/cip1蛋白的表达没有相关性(P>0.05)。结论:FHIT基因可能成为早期诊断膀胱移行细胞癌的指标。p21waf1/cip1基因可能成为估计膀胱移行细胞癌的恶性程度及肿瘤侵袭性、预后的指标。FHIT基因在膀胱移行细胞癌中的作用机制可能与p21waf1/cip1基因没有关系。     

Effect of p21waf1/cip1 transgene on radiation induced apoptosis in T cells.

The cyclin kinase inhibitor p21WAF1/Cip1 is upregulated by the tumor suppressor p53. While p21 is central for the G-1 arrest mediated by p53, it is still unclear if p21 also functions as a downstream effector of p53 dependent apoptosis. Apoptosis induced by DNA damage but not dexamethasone is p53 dependent in thymocytes. To investigate the physiological role of p21 in apoptosis, we have generated transgenic mice in which the p21 transgene is targeted for restricted expression in the T cell lineage. Thymocytes from p21 transgenic mice were hypersensitive to cell death induced by DNA damaging agents such as ionizing radiation and UV, but not be dexamethasone. Irradiated p21 transgenic thymocytes had approximately twofold more apoptotic cells as compared to irradiated age matched littermate control mice. Radiation induced death is comparable in thymocytes from p21 + Bcl2 + double transgenic mice and age matched littermate controls, indicating that the Bcl2 transgene rescues the radiation hypersensitivity imposed by p21. However, thymocytes from p53-/- mice even when they expressed the p21 transgene, were resistant to death induced by radiation. Together these results show that thymocytes from p21 transgenic mice are hypersensitive to radiation induced programmed cell death and suggest that the radiation hypersensitivity of p21 transgenic thymocytes involves p53 dependent pathway and signals in addition to p21.     

Effects of p21cip1/waf1 overexpression on growth, apoptosis and differentiation in human colon carcinoma cells

The cyclin-dependent kinase inhibitor p21cip1/waf1 negatively regulates the progression of cell cycle and the potential usefulness of p21cip1/waf1 gene is proposed in gene therapy. However, studies have demonstrated a protective role of p21cip1/waf1 against apoptosis and little is known about effects of ectopic expression of p21cip1/waf1 on differentiation of colon cancer cells. In the present study, we found diffuse p21cip1/waf1 expression in only a few clinical samples of colorectal cancer with wild-type p53 gene. To explore the role of p21cip1/waf1 in cell growth, apoptosis and differentiation, we constitutively overexpressed p21cip1/waf1 in HT29 colon carcinoma cells. Ectopic overexpression of p21cip1/waf1 was associated with inhibition of CDK2-associated kinase activity, indicating the functionality of the introduced p21cip1/waf1 gene. Overexpression of p21cip1/waf1 caused an appreciable growth inhibition in monolayer and soft agar cultures and it significantly reduced sodium butyrate- but not 5-fluorouracil-induced apoptosis. p21cip1/waf1 overexpressing cells exhibited marked decrease of intestinal differentiation when assayed with intestinal alkaline phosphatase. Our findings suggest that introduction of p21cip1/waf1 gene into colon cancer cells may be useful for inhibiting cell growth but caution should be taken regarding the increased resistance to certain apoptosis-inducing agents and dysregulation of endogenous p21cip1/waf1-mediated differentiation process.