食管鳞癌自发细胞凋亡和核增殖抗原p53关系的研究

目的通过对食管鳞癌组织中凋亡细胞的原位观察和核增殖抗原（ＰＣＮＡ）的表达状态的研究,探讨不同增殖情况的食管鳞癌组织细胞凋亡情况,同时研究ｐ５３基因的突变和ｐ５３蛋白的表达对食管鳞癌组织细胞凋亡的影响。方法３０例术前未经任何治疗的食管鳞癌手术标本,分别进行凋亡细胞的原位检测（ＴＵＮＥＬ）、ＰＣＮＡ、ｐ５３蛋白的免疫组化染色和ｐ５３基因５,６,７,８外显子ＰＣＲ－ＳＳＣＰ检测。结果不同增殖能力的癌组织中,细胞凋亡的程度差异有显著性（Ｐ＜０．０５）。ｐ５３基因突变组中,增殖程度高与增殖程度低者之间癌细胞凋亡的程度差异有显著性（Ｐ＜０．０５）。ｐ５３基因突变组与非突变组、ｐ５３蛋白表达阳性组与非阳性组癌细胞凋亡的程度差异无显著性（Ｐ＞０．０５）。结论食管鳞癌组织中,细胞增殖活跃则自发性凋亡也相应增多     

食管鳞状细胞癌组织中P53蛋白的积聚及其临床意义

采用免疫组化方法，以Ｄ０－７单克隆抗体检测５７例食管鳞状细胞癌组织中Ｐ５３蛋白的表达，结果：４８例癌组织中有不同程度的Ｐ５３蛋白积聚，阳性率为８４．２１％，从２１例有淋巴结转移的病例共取得６１个转移癌淋巴结，其中５８个淋巴结中转移癌呈Ｄ０－７阳性，与其原发瘤相比，阳性程度增高，将癌组织中Ｐ５３蛋白积聚情况与人临床病情分期进行分析，分析结果显示晚期病人癌组织中Ｐ５３蛋白聚积有增加趋势。除此之外，对５     

胸腺癌中p53蛋白过表达对癌细胞增殖和凋亡的影响

目的：探讨胸腺癌中ｐ５３蛋白过表达对癌细胞增殖和凋亡的影响。方法：收集手术切除的胸腺标本２０例,用抗ｐ５３蛋白（ＤＡＫＯ ＤＯ－７）和抗增殖细胞核抗原（ＰＣＮＡ,ＤＡＫＯ ｃｌｏｎｅ ＰＣ１０）的单抗,采用ＬｓＡＢ免疫组化法染色。以平均每１００个癌细胞中ｐ５３阳性细胞数为ｐ５３表达率,阳性ｐ５３蛋白细胞数超过２０％者称为ｐ５３蛋白过表达。以平均每１００个癌细胞中ＰＣＮＡ阳性细胞数为增殖指数（ｐｒｏ     

食管癌及癌前病变组织P53蛋白表达的研究

对１９７例食管内窥镜活检组织中Ｐ５３蛋白的免疫组化分析表明，约３８．８％的食管轻度不典型增生，５２．０％的中重度不典型增生，６１．１％的原位癌和６２．５％食管浸润癌组织中有高表达的Ｐ５３蛋白。正常食管鳞状上皮和慢性皮炎症食管上皮中很少有Ｐ５３蛋白聚积，对１４例食管癌病人癌及癌旁不典型增生组织中Ｐ５３蛋白的免疫组化分析表明，９例Ｐ５３蛋白高表达的食管癌中，８例癌旁不典型增生组织也有高表达的Ｐ５３基因     

免疫组化法检测鼻咽病变组织中PCNA和P53的意义

本文报道采用免疫组化ＡＢＣ法检测了８０例鼻咽活检组织的ＰＣＮＡ和Ｐ５３蛋白表达情况，结果显示：（１）ＰＣＮＡ和Ｐ５３表达的阳性率在鼻咽癌分别为９４．２％和８４．６％，异型性改变为７０．０％和２８．８％，单纯性增生为３．９％和阴性，正常上皮均为阴性；（２）ＰＣＮＡ和Ｐ５３表达的细胞阳性强度和阳性细胞数量与鼻咽癌的组织类型无关，但ＰＣＮＡ和Ｐ５３表达的细胞阳性强度及Ｐ５３表面的阳性细胞数量与肿瘤的间变     

胃粘膜异型增生上皮p53蛋白表达的临床意义

采用免疫组化方法，对２４例常规胃镜活检的胃粘膜异型增生上皮的ｐ５３蛋白表达进行检测，并做１～２４个月的随访研究。结果表明，ｐ５３蛋白阳性的异型增生上皮的癌检出率为５７．１％（４／７），其中２例为早期胃癌，而ｐ５３阴性的异型增生上皮的癌检出率为１１．８％（２／１７），两者有显著性差异（Ｐ＜０．０５）。结果提示，胃粘膜上皮ｐ５３蛋白表达是一项恶性生物学指标，可作为早期诊断及鉴别诊断胃癌的标志     

P~(21)、P~(53)蛋白在胃癌及胃粘膜异型增生中的表达

本研究对４２例胃癌病例及４３例胃粘膜异型增生病例行Ｒａｓ癌基因Ｐ２１蛋白、抑癌基因Ｐ５３蛋白单克隆抗体免疫组化研究，结果发现胃癌组织中Ｐ２１阳性率为５４．７％，并与癌组织分化程度呈正相关。Ｐ５３蛋白阳性率为４２．８％，与癌组织分化程度呈负相关，说明胃癌组织中Ｐ２１蛋白及Ｐ５３蛋白的表达与其发生发展有重要关系，并与胃癌的生物学行为有关，在异型增生病例中Ｐ２１及Ｐ５３阳性多为重度病例，说明癌基因的激活与抑癌基因的突变在异型增生发生发展及癌变过程中起着重要作用。     

喉粘膜上皮异型增生p53、H-ras过度表达与预后的关系

目的：探讨喉粘膜上皮异型增生ｐ５３、Ｈ－ｒａｓ过度表达与预后的关系。方法：应用免疫组化ＡＢＣ法对６３例喉粘膜上皮异型增生（经随访其中４０例恶变为癌），５６例喉鳞癌和７例喉正常组织进行检测。结果：ｐ５３蛋白阳性率在喉粘膜上皮异型增生恶变组为５０．０％，未恶变组为１７．４％，喉癌组为５１．８％，正常组织无表达。恶为组与未恶变组间，未恶变组与喉癌组间的Ｐ５３蛋白表达差异有统计学意义（分别为Ｐ〈０．０５和     

nm23和p53蛋白在喉鳞癌组织中表达的意义

应用免疫组化技术，了３２例喉鳞癌组织及１２社异型增生组织的ｎｍ２３和ｐ６３蛋白的表达情况。结果发现：ｎｍ２３与ｐ５３在喉鳞癌组织的阳性一表达率分别为８７．５０％及８４．３８％。在癌旁异型增生组织阳性表达率分别为１００％和２０％。ｎｍ２３的表达与喉鳞癌的组织学分级有关，Ｐ５３的表达与喉鳞癌组织学分级无关，但Ｐ５３的强阳性表达预示预后不良。     

P^21,P53蛋白在胃癌及胃粘膜异型增生中的表达

本研究对４２例胃癌病例及４３例胃粘膜异型增生病例行Ｒａｓ癌基因Ｐ＾２１蛋白、抑癌基因Ｐ＾５３蛋白单克隆抗体免疫组化研究，结果发现胃癌组织中Ｐ＾２１阳性率为５４．７％，并与癌组织分化程度呈正相关。Ｐ＾５３蛋白阳性率为４２．８％，与癌组织分化程度呈负相关，说明胃癌组织中Ｐ２１蛋白及Ｐ＾５３蛋白 表达与其发生发展有重要意义，并与胃癌的生物学行为有关，在异型增生病例中Ｐ＾２１及Ｐ６５３阳性多为重度病例，说     

食管鳞状细胞癌组织中P53蛋白的积聚及其临床意义

采用免疫组化方法，以DO—7单克隆抗体检测57例食管鳞状细胞癌组织中P53蛋白的表达。结果：48例癌组织有不同程度的P53蛋白积聚，阳性率为84．21％（48／57）。从21例有淋巴结转移的病例共取得61个有转移癌淋巴结，其中58个淋巴结中转移癌呈DO－7阳性，与其原发癌相比，阳性程度增高。将癌组织中P53蛋白积聚情况与病人临床病情分期进行比较，分析结果显示晚期病人癌组织中P53蛋白聚积有增加趋势。除此之外，对57修例食管鳞状细胞癌进行了PCNA染色，绝大多数P53阳性的癌细胞也呈PCNA阳性，二者呈明显的正相关，表明有P53蛋白积聚的癌细胞具有更活跃的增殖能力。本文结果提示，食管鳞状细胞癌中P53蛋白积聚与肿瘤演进有一定关系。     

Immunohistochemical detection of p53 and Bcl-2 in colorectal carcinoma: no evidence for prognostic significance.

To evaluate the prognostic significance of immunohistochemically detected p53 and Bcl-2 proteins in colorectal cancer, tissue sections from 238 paraffin-embedded colorectal carcinomas were immunostained for p53 (MAb DO-7 and CM-1 antiserum) and Bcl-2 (MAb Bcl-2:124). Staining patterns were assessed semiquantitatively and correlated with each other and with sex, age, tumour site, Dukes'' classification, tumour differentiation, mucinous characteristics, lymphocyte and eosinophilic granulocyte infiltration, and patient survival. In our series, 35% of carcinomas showed no nuclear staining and 34% (DO-7) to 40% (CM-1) showed staining in over 30% of tumour cell nuclei. A majority of carcinomas that had been immunostained with CM-1 showed cytoplasmic staining, but this was not observed with DO-7. With respect to Bcl-2, 51% of tumours were completely negative, 32% displayed weak and 15% moderate staining; only 3% showed strong positive staining. No evidence was found for reciprocity between Bcl-2 expression and nuclear p53 accumulation. From 13 cases containing tumour-associated adenoma, four were Bcl-2 negative in premalignant and malignant cells, in another four cases these cells showed similar staining intensities and in the remaining cases only the malignant colorectal cells were Bcl-2 negative. Therefore, our data indicate that Bcl-2 is dispensable in the progression towards carcinoma. Except for an association between nuclear p53 accumulation and mucinous tumours (P = 0.01), no significant correlation was found between the clinicopathological parameters mentioned above and immunostaining pattern of (nuclear or cytoplasmic) p53 or Bcl-2.     

我国不同地区食管癌中p53蛋白积聚的比较观察

作者收集了中国食管癌三个高发区（郑州、大原、汕头）和一个低发区（广州）的食管鳞癌手术切除标本，福尔马林固定，石蜡包埋，采用CM－1和Do－7作一抗，用免疫组化LSAB法对其进行检测，在四个不同地区p53蛋白积聚率无差异（P＞0．05），阳性率为62．8％（Do－7）。因此，食管鳞癌中p53蛋白积聚是一常见现象，在不同地区，p53的改变在该肿瘤形成过程中的作用是类同的。作者还发现PCNA（Proliferatingcellnuclearantigen）的表达与p53蛋白积聚的分布规律一致，两者阳性等级密切相关（P＜0．01），说明有p53蛋白积聚的肿瘤细胞是处于细胞增殖周期中，可能与肿瘤的预后有一定的关系。     

p53 protein alterations in human testicular cancer including pre-invasive intratubular germ-cell neoplasia.

Expression of the p53 oncoprotein was examined in a wide range of primary human testicular germ-cell tumours using a new mouse monoclonal antibody (MAb) BP53-11 raised and characterized in this study, in parallel with a polyclonal rabbit antiserum CM-1. Immunohistochemistry on paraffin sections showed positive nuclear reaction in at least a fraction of malignant cells in 90 (84%) out of 107 cases studied. Aberrant accumulation of the p53 protein was found among testicular tumours of all major histological types, although generally a higher percentage of positive cases and a higher proportion of p53 over-expressing nuclei within individual lesions was observed in embryonal carcinomas when compared with seminomas. The typical heterogeneous staining pattern characteristic of histological specimens was also found in a cultured cell line derived from a human embryonal carcinoma. In contrast to immunohistochemically undetectable levels in normal testes and morphologically normal tissue areas in the tumour-bearing testes, the accumulation of the p53 protein was clearly identified in a high proportion (59% of cases) of the pre-invasive lesions with positive atypical intratubular germ cells often found in the tissue adjacent to invasive tumours. Altered expression of the p53 protein is therefore a unifying feature of the majority of invasive male germ-cell tumours and the change resulting in high levels of p53 appears to be a relatively early step in the human testicular cancer pathogenesis.     

Circulating p53 antibodies, p53 gene mutational profile and product accumulation in esophageal squamous-cell carcinoma in India

Esophageal cancer (EC) in the Indian population presents in advanced stages with poor prognosis and warrants the identification of a non-invasive marker for early detection and better prognostic assessment. We have previously reported high prevalence of p53 protein accumulation in esophageal squamous-cell carcinomas (ESCCs). The present study was designed to determine (i) if esophageal cancer patients elicit a humoral immune response to intra-tumoral p53 protein accumulation and (ii) their relationship with p53 gene mutations. The goal was to compare the cellular events, p53 protein accumulation and gene mutations with the presence of serum anti-p53 antibodies (p53-Abs) and to assess the utility of serological p53-Ab analysis as a surrogate marker for p53 alterations in esophageal cancer. A high prevalence of circulating p53-Abs was observed in 36 of 60 (60%) ESCC patients. In a subset of 44 ESCCs, exons 5-9 of the p53 gene were examined for mutations by PCR and direct sequencing of PCR products. Mutational data have been correlated with p53-Abs and p53 protein accumulation in ESCCs. Circulating p53-Abs in ESCC patients were significantly associated with intra-tumoral p53 protein accumulation (p=0.0005). A strong correlation observed between humoral immune response against p53 protein, missense gene mutations and protein accumulation warrants the application of serological p53-Abs as a non-invasive surrogate marker in screening high-risk populations for early detection of malignancy.     

Comparison between p53 staining in tissue sections and p53 proteins levels measured by an ELISA technique.

We studied 51 paired samples of tissue sections and cytosol extracts from patients with breast cancer. A very high affinity monoclonal antibody to human p53 protein, DO-1, and polyclonal serum CM-1 to p53 protein were used for two site ELISA assays and CM-1 was used for immunohistochemistry to detect p53 protein accumulation in breast cancer samples. Eighteen carcinomas were positive for p53 by tissue staining and ELISA assay. Nineteen tumours were negative by ELISA and immunohistochemistry, and 14 cases with low levels of positive staining by immunohistochemistry were negative by the ELISA assay. A statistically significant correlation has been found between the degree of staining and the amount of p53 protein measured by ELISA (Pearson''s correlation coefficient r = 0.59, P < 0.00001). Our ELISA assay offers an alternative approach to evaluating the p53 status of breast biopsy material, using cytosol extracts routinely prepared for steroid hormone receptor assays. This assay should also be of general application to other situations where the level of p53 protein needs to be determined.     

Alterations in MDM2 expression in esophageal squamous cell carcinoma: Relationship with p53 status

In view of the significance of MDM2 as a regulator as well as critical target of wild type p53, this study was undertaken to determine the alteration in MDM2 expression in esophageal squamons cell carcinoma (ESCC) and its relationship to clinicopathological parameters as well as p53gene and protein status. Immunohistochemical analysis of MDM2 and p53 proteins on paraffin embedded sections from 64 surgically resected ESCCs and matched histologically normal tissues showed overexpression of MDM2 protein in 23/64 (36%) ESCCs, while the histopathologically normal esophageal tissues did not show detectable level of MDM2 immunoreactivity. Interestingly, MDM2 /p53 + phenotype was observed in 37/64 (58%) cases. None of the cases with p53 mis-sense mutations (12/30, 40%) showed detectable level of MDM2 protein. Missense p53 mutations were significantly associated with discordant p53 + /MDM2 immunophenotype (p= 0.004). The most intriguing feature of the study was accumulation of MDM2 in the absence of detectable p53 in 11% of and overexpression of MDM2 and p53 in 25% of ESCCs, suggesting a p53-independent role for MDM2 in a subset of tumors. These results underscore the involvement of MDM2 in p53-dependent and -independent pathways in the pathogenesis of esophageal cancer in the Indian population.     

P53 expression in esophageal dysplasia - a possible biomarker for carcinogenesis of esophageal squamous-cell carcinoma

The tumor suppressor gene product p53 has been detected in a high percentage of esophageal squamous cell carcinoma. To evaluate the role of this protein in carcinogenesis, we examined the p53 overexpression both in esophageal dysplasia and in esophageal squamous cell carcinoma in the same patients. Using anti-p53 antibodies pAb1801 and CM-1, we analyzed immunohistochemically 36 dysplastic lesions from 36 patients with esophageal cancer. Nuclear p53 was detected in 14 of 36 dysplasias (39%). From mild to moderate to severe dysplasia, p53 positivity showed tendency to increase in number. Seventeen of the 36 squamous cell carcinomas showed p53 expression (47%). There was a significant concurrent p53 expression in esophageal dysplasia and its related squamous cell carcinoma (p=0.00345). These results indicate that p53 mutation is closely associated with the initiation of this cancer.     

抑癌基因p53在不同地区食管癌中的表达及临床预后分析

目的：比较中国食管癌高，低发区p53蛋白的积聚，燕对p53蛋白积聚与肿瘤细胞增殖及临床预后的关系进行探讨。方法：用免疫组化LSAB法检测高发区（河南）43例食管癌和低发区（广东）40例食管癌p53蛋白及增殖细胞核抗原的表达。结果：p53蛋白阳性率分别是60．5％（河南）和57．5％（广东）。阳性率差异无显著性（P＞0．05）。p53蛋白表达无论在高，低发区均与PCNA的表达密切相关（P＜0．001）。结论：食管癌不同的高，低发区中p53蛋白的各聚是类同的。大多数p53蛋白积聚的肿瘤细胞是处于细胞增殖周期内，可能成为食管癌恶性程度的指标之一，将为临床分析预后提供客观依据。     

Unusual profile and high prevalence of p53 mutations in esophageal squamous cell carcinomas from northern Iran

Over 15,000 human tumor p53 mutations have been recorded in the scientific literature, including over 700 mutations in esophageal tumors. There are no data on p53 mutations in esophageal cancer patients from Iran yet; however, this country experiences one of the highest cancer mortality rates in the world for esophageal squamous cell carcinomas (ESCCs). The causes of this high cancer burden in Iran remain obscure and do not appear to be related to tobacco and alcohol consumption, the two major risk factors identified in Europe and North America. Because molecular analysis of tumors can provide clues to endogenous or environmental factors contributing to high cancer risk, we examined 74 Iranian ESCCs for the presence of mutations in exons 5-8 of the p53 gene by PCR and direct sequencing. Forty-eight of the 74 tumors (65%) had one or more p53 gene point mutations, including 5 patients with two or more mutations and one with a tandem mutation in codon 242. Surprisingly, over one-third of the 54 mutations we identified were transitions at CpG sites (20 of a total of 54 mutations, or 37%), a class of mutation that is significantly less common (16% of mutations) in the compilation of ESCC mutations from other countries (chi2 statistic, P < 0.0002), whereas transversions, which the literature shows to be common in ESCCs from non-Iranian patients, were infrequent in the tumors we examined here. Elevated levels of cyclooxygenase-2 and inducible nitric oxide synthase were observed in 74 and 91%, respectively, of tumors from Tehran as determined by immunohistochemistry, and high COX-2 expression correlated significantly with the presence of a p53 mutation in the tumor. Mediators of the inflammatory response in esophageal mucosa, perhaps in conjunction with specific dietary or cultural practices in Iran, may contribute importantly to the p53 mutation load in Iranian ESCC patients.