Clinical importance of correlations between p53 immunoreactivity and clinicopathological parameters in lung carcinoma

Many studies have revealed the frequency of p53 abnormalities in lung cancer. However, clinico-pathological studies of p53 abnormalities have yielded conflicting results. We examined the p53 immunoreactivity and studied the correlations of p53 status and clinicopathological parameters in 76 primary lung cancers. By using DO-7 antibody, different degrees of p53 immunoreactivity was detected in 8 of 30 small cell lung cancer (SCLC, 26.6%) and 22 of 46 non-small cell lung cancer (NSCLC, 47.8%), 6 of 19 adenocarcinoma, 16 of 27 epidermoid carcinoma cases. In the whole group, no correlation was detected between the p53 status and the histological types of tumor, local tumor invasion, nodal status, and distant metastasis and patient characteristics, such as age, gender or smoking habit. P53 status was also found to have no effect on survival. However, in the NSCLC group, there was a significantly higher p53 immunoreactivity in well- and moderately-differentiated tumors (p<0.05). Patients with p53 immunoreactivity had a poor therapeutic response in the whole group. We concluded that, although p53 immunreactivity may be found in NSCLC, this does not correlate with clinicopathological parameters except therapeutic response. In SCLC p53 immunreactivity can be negligible.     

Association between p53 mutation and clinicopathological features of non-small cell lung cancer

Genetic alterations in exons 5-8 of the p53 gene, determined by single-strand conformation polymorphism and sequencing analyses, and the clinicopathological characteristics of 108 patients with non-small cell lung cancer were compared. Mutations in this gene were found in 37 of the 108 patients (34%): in 30% (23/76) of those with adenocarcinomas, 46% (12/26) of those with squamous cell, 33% (1/3) of those with large cell and 33% (1/3) of those with adenosquamous carcinomas. No associations between the incidence of p53 mutations and the histological or cytological subtypes of adenocarcinomas were found. The analysis of types of mutations, however, showed that GC transversion was relatively common in papillary and clara subtypes, whereas it accounted for only 17% at most of p53 mutations in tubular and bronchial surface epithelial cell subtypes of adenocarcinomas. Univariate analyses revealed that large tumor size, high nodal stage and positive vascular invasion of non-small cell lung cancers, and high nodal stage and high-grade nuclear atypia of adenocarcinomas were associated significantly with p53 mutations. Multivariate analyses showed that the tumor sizes of non-small cell lung cancer correlated with p53 mutations with marginal significance (P = 0.099) whereas nuclear atypia of adenocarcinomas correlated significantly (P = 0.028). No differences between the overall or relapse-free survival rates of patients with and without p53 mutations in non-small cell lung cancers or adenocarcinomas were found. These findings indicate that p53 mutations in adenocarcinomas of the lung are associated with the malignant phenotype of tumor cells, but not with patient survival.      

Over-expression of p53-protein and cigarette-smoking in bronchial-carcinoma

A recessive gene on chromosome 17 encodes a protein, known as p53, which normally acts to regulate the cell cycle, its mutation and over-expression being amongst the commonest genetic abnormalities in human malignant neoplasms. As detected by immunolabelling using the anti-p53 protein antibodies PAb1801, DO7 and CM1, over-expression was demonstrable in 13 of 59 tissue biopsy specimens from a series of patients with newly-diagnosed primary bronchial carcinoma from whom accurate data on smoking history had been obtained prior to bronchoscopy. There was a statistically significant relationship between over-expression and total exposure to cigarette smoke (p53-positive, median 46 pack-years; p53-negative, median 32 pack-years; P<0.001) and between over-expression and intensity of exposure (p53-positive, median 20 cigarettes/day; p-53 negative, median 14 cigarettes/day; P<0.001), but no difference between the two groups in terms of total duration of this exposure (p53-positive, median 47 y; p53-negative, median 46 y). These data confirm findings of previous studies suggesting a possible link between cigarette smoke and those derangements of the structure or function of the p53-encoding gene which lead to its over-expression by malignant bronchial tumours. They support, in addition, a specific mutagenic role for the chemical carcinogens it contains. Over-expression of p53 did not appear to influence survival.     

p53 gene aberrations in non-small-cell lung carcinomas from a smoking population.

We examined 46 non-small-cell lung carcinomas (NSCLCs) for the presence of p53 mutations in exons 4-9, positive p53 immunostaining and loss of heterozygosity (LOH) in the TP53 locus. p53 mutations were detected in 13 tumour samples (28.3%), whereas overexpression of the p53 protein was found in 30 of 45 (67%) samples. Allelic loss was found in 9 of 38 (23.6%) informative cases. The statistical analysis revealed no significant correlation between p53 mutations and clinicopathological data, although mutations appear to occur more frequently in squamous cell carcinomas (7 of 18) than in adenocarcinomas (2 of 15). All but three individuals in this study group smoked. In contrast to previous reports, we found a higher prevalence of GC-->AT transitions than of GC-->TA transversions, as expected in a smoking population. A trend was found between p53-positive immunostaining and a history of heavy smoking (76-126 pack-years) and was inversely correlated with allelic deletion (LOH) at the TP53 locus. Eight of the 12 NSCLCs containing p53 mutations also had concomitant p53 overexpression, and it is of specific note that three of the four tumours containing p53 ''mutations'' with no overexpression of the p53 protein had either insertions or deletions in the p53 gene. No correlation was found between p53 mutations and fractional allele loss or ras mutations. p53 mutations in this Merseyside population in the UK do not appear to be as common as in other reports for NSCLC and exhibit predominance of GC-->AT transitions preferentially at non-CpG sites, suggesting that other carcinogens in addition to those in tobacco smoke may be involved in NSCLC in the Merseyside area of the UK.     

Immunohistochemical detection of H-ras protooncoprotein p21 indicates favorable prognosis in node-negative breast cancer patients.

We tested primary breast carcinoma tissues from 297 patients using a monoclonal antibody (clone 235-1.7.1.) for the expression of p21(ras). 58% of tumors were p21(ras)-positive. When calculated in a univariate fashion, p21 expression correlated with proliferation activity (proliferating cell nuclear antigen) only. Using the log-rank test (median observation time 94 months) a significantly worse prognosis (disease-free survival, overall survival) relation was found with larger tumors, nodal involvement, anaplasia, rising proliferation activity, and lack of steroid receptors. Detection of p21(ras) correlated with a more favorable prognosis but only in node-negative patients. Stepwise correlation according to the Cox hazard model ranked p21 expression as a most significant predictor of prognosis second only to nodal status. These data suggest that detection of p21(ras) indicates the presence of a parameter which may act as tumor suppressor and benefit patients' survival.     

晚期非小细胞肺癌组织中ERCC-1、Metallothionein、p53表达与铂类耐药性及预后的相关性分析

背景与目的晚期非小细胞肺癌含顺铂方案化疗敏感性分子预测指标尚处于探索阶段。本研究分析晚期非小细胞肺癌石蜡包埋组织中错配切除修复蛋白(excisionrepaircrosscomplement-1,ERCC-1)、金属硫蛋白(metallothionein,MT)、p53蛋白与顺铂化疗敏感性的关系,并观察这些指标与病人生存的关系。方法收集1994年1月-2001年12月在本院经病理组织学诊断的用含铂方案(Gemzar+DDP或NVB+DDP)化疗的初治晚期非小细胞肺癌患者的病理标本,采用免疫组化方法检测ERCC-1、MT、p53的表达,并将病人的疗效和生存资料与上述指标的表达进行比较。样本率的比较用χ2检验,用Kaplan-meier法和Cox回归进行生存分析。结果本组51例标本中,ERCC-1阳性率为42.8%,MT阳性率为57.5%,p53阳性率为37.8%。化疗有效率在ERCC-1、MT、p53阴性组和阳性组分别为33.3%、35.3%、21.4%和16.7%、27.3%、35.3%(即分别为1.99倍,1.29倍,0.61倍),但均未达统计学差异(P>0.05),ERCC-1,MT均阴性组有效率为37.5%,均阳性组有效率为14.3%,前者为后者的2.6倍,但未达统计学差异(P>0.05)。平均生存期ERCC-1、MT、p53蛋白阴性组和阳性组分别为621.03天/523.14天,556天/479天,599.64天/416.60天。Kaplan-meier分析显示三者过度表达均提示预后不良,但Cox回归分析显示只     

p53 status and prognosis in stage I-IIIa non-small cell lung cancer

To investigate the role of p53 abnormalities in predicting the survival of patients with non-small cell lung cancer (NSCLC), polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and immunohistochemical analyses were performed on 74 and 67 tumor samples, respectively, from patients with pathological stage I-IIIa NSCLC. An abnormally migrating SSCP band was observed in 21 of 74 (28%) tumor specimens. DNA sequence analysis revealed 23 intragenic mutations including 3 small deletions and 20 point mutations. Immunohistochemical analysis using the DO-7 monoclonal antibody showed abnormal expression of p53 in 27 of 67 (40%) patients. The concordance rate between immunohistochemical and PCRSSCP analyses was 73% (49/67) in this study. Univariate and multivariate analyses demonstrated that abnormal expression of p53 may be associated with prolonged survival (p=0.0997 and 0.0099, respectively). In contrast, no relationship was observed between p53 mutation and overall survival (0.6968). These results suggest that p53 status and the survival outcome changes between immunohistochemical and mutational analyses in stage I-IIIa NSCLC.     

ras p21和p53的表达与尿路上皮肿瘤临床预后的关系

目的 研究ｒａｓ ｐ２１和ｐ５３的表达与尿路上皮肿瘤临床与预后的关系。方法 采用免疫组织化学ＡＢＣ法检测６６例尿路上皮肿瘤ｒａｓ ｐ２１和ｐ５３的表达。结果 ｒａｓ ｐ２１阳性表达４０例（６０．６％）,ｐ５３阳性表达３６例（５４．５％）,ｒａｓ ｐ２１和ｐ５３共同阳性表达２７例（４０．９％）;ｒａｓ ｐ２１和ｐ５３阳性表达随肿瘤病理分级和分期的上升而增强,与肿瘤复发及预后呈正相关性（Ｐ〈０．０１）     

Clinical and pathological associations with p53 tumour-suppressor gene mutations and expression of p21WAF1/Cip1 in colorectal carcinoma.

Inactivation of the p53 tumour-suppressor gene is common in a wide variety of human neoplasms. In the majority of cases, single point mutations in the protein-encoding sequence of p53 lead to positive immunohistochemistry (IHC) for the p53 protein, and are accompanied by loss of the wild-type allele. Recently, the WAF1/Cip1 gene was identified as one of the genes induced by wild-type p53, and increased expression of p21WAF1/Cip1 has been found to reflect the status of the p53 tumour-suppressor pathway. We investigated the inactivation of p53 in a relatively small, but well-characterised, group of 46 colorectal carcinomas that were previously studied for allelic alterations, ras oncogene mutations and DNA aneuploidy. Alterations in p53 were identified by IHC, loss of 17p and DNA sequence analysis of exons 5-8, whereas p21WAF1/Cip1 protein expression was determined by IHC. p53 mutations were identified in 19 of the 46 tumours (41%), whereas positive IHC for p53 was found in 21 of the 46 tumours (46%). Positive IHC for p21WAF1/Cip1 was detected in 16 of 42 cases (38%). We found no relationship between p21WAF1/Cip1 staining and p53 protein expression or p53 mutational status. Inactivating mutations in the p53 gene correlated with LOH at 17p but not with LOH at 5q or 18q, Dukes'' stage, tumour grade or DNA ploidy. There was a higher survival rate independent of Dukes'' stage in the group with no alterations in p53 compared with those with evidence of dysfunction of p53, but the difference was not statistically significant. We conclude that inactivation of p53 and altered expression of p21WAF1/Cip1 are common in colorectal carcinoma but do not correlate with each other or with the clinical or pathological parameters investigated.     

Comparative analysis of p16/CDKN2, p53 and ras gene alterations in human non-small cell lung cancers, with and without associated pulmonary asbestosis

To investigate genetic abnormalities in human non-small cell lung carcinomas (NSCLC) associated with pulmonary asbestosis as compared with nou-asbestos linked lung cancers, twenty-nine primary non-small cell lung carcinomas (NSCLC) were examined for genetic abnormalities of p16/CDKN2, p53 and ras genes by single-strand conformation polymorphism analysis of polymerase chain reaction products (PCR-SSCP) and direct sequencing. Ten specimens were obtained from autopsies in which concurrent pulmonary asbestosis was present, while 19 samples were surgical specimens from asbestosis-free patients. K-ras mutations were detected in 10% each of the cancers from both asbestosis and non-asbestosis cases. p16/CDKN2 deletions or mutations and p53 aberrations were demonstrated in 20% and 10% of tumors from asbestosis cases, whereas, 32% and 21% of the cancers, respectively, from asbestosis-free patients were positive. In conclusion, it is suggested that the enhancement of neoplasia in the lung by asbestos is not dependent on suppression of p16/CDKN2 and p53 or ras activation and therefore, that asbestosis may activate alternate tumorigenic pathways in the development of NSCLC.     

P糖蛋白在胰腺癌中的表达及其与p21^ras和p53蛋白表达的关系

为了解Ｐ糖蛋白在原发和复发胰腺癌组织中的表达及其临床意义，并分析其表达与Ｐ２１＾ｒａｓ和Ｐ５３蛋白表达的关系，采用免疫组化泽４８例原发胰腺导管癌和１５例复发胰腺癌的石蜡包埋标本进行Ｐｇｐ，ｐ２１＾ｒａｓ蛋白和ｐ５３蛋白单抗的免疫组化染色。     

p21、p185、p53蛋白在卵巢浆液性肿瘤中表达的临床意义

目的：探讨ｒａｓ、ｎｅｕ、ｐ５３基因在肿瘤演进过程中的作用。方法：采用链蛋白生物素免疫组化法（ＬＳＡＢ）检测了ｒａｓ、ｎｅｕ、ｐ５３基因产物ｐ２１、ｐ１８５、ｐ５３蛋白在卵巢浆液肿瘤中的表达情况。结果：三种蛋白在良性、交界性和恶性肿瘤组织中阳性率呈递增趋势，联合表达率亦呈递增趋势，差异显著（Ｐ＜０．０５），ｐ２１表达与患者生存期有关，过度表达组低于非过度表达组（Ｐ＜０．０１）。结论：联合检测ｐ２１、ｐ１８５、ｐ５３蛋白在卵巢浆液性肿瘤组织中的表达情况，对该肿瘤的诊断及预后判定具有重要参考价值     

端粒酶与p53、p16基因在非小细胞肺癌中的表达及其意义

目的研究端粒酶与p53、p16基因在非小细胞肺癌(NSCLC)中的表达及其意义.方法应用SP法免疫组化技术和TRAP法分别检测40例肺癌组织中端粒酶和p53、p16基因的表达.结果端粒酶及p53、p16基因在NSCLC中的阳性率分别为92.5%、37.5%和66.7%;p53基因阳性表达率与NSCLC及病理分期显著相关(P＜0.05).p16基因阳性表达率与性别、年龄、病理分期、吸烟史、组织学类型和分化程度无显著相关(P＞0.05).端粒酶活性与年龄呈负相关,并与p53基因表达情况和病理分期有关(P＜0.05).结论端粒酶和p53基因的表达与NSCLC的病期进展有关.     

肺鳞癌细胞p~(53)蛋白及ras p~(21)的免疫组织化学定量研究

应用免疫组织化学方法检测２２例肺鳞癌组织标本的ｐ５３及ｒａｓｐ２１表达，用显微图像分析仪测定阳性细胞百分率Ｐ／Ａ（％）和平均光密度（ＡＯＤ），并采用阳性水平指数（ｐｏｓｉｔｉｖｅｌｅｖｅｌｉｎｄｅｘ，ＰＬＩ）作为免疫组化反应水平指标，对癌基因蛋白进行比较研究。结果显示：ｐ５３表达阳性率为５０％，ｒａｓｐ２１表达阳性率为６３６％。ｐ５３蛋白表达在非角化型鳞癌组高于角化型鳞癌组；ｒａｓｐ２１的表达随组织分化程度的增高而增高，ｒａｓｐ２１与ｐ５３的表达水平无直线相关关系。     

p53 gene mutations in non-small-cell lung cancer cell lines and their correlation with the presence of ras mutations and clinical features.

We screened 77 non-small-cell lung cancer (NSCLC) cell lines for mutations of the p53 gene using a single-strand conformation polymorphism (SSCP) assay. We found that 57 cell lines (74%) had mutations of the p53 gene. Three cell lines had a deletion of the p53 gene. Of the remaining 54 cell lines, 49 cell lines were sequenced and 52 mutations were confirmed. In contrast to previously published p53 mutations in other human tumors, the p53 gene mutations in NSCLC were diverse with regard to the location and nature of the mutations. The region corresponding to codons 144-166, which is outside the evolutionarily conserved regions, was a frequent site of p53 gene mutations in NSCLC. The presence of a p53 gene mutation was not associated with age, sex, histological types, culture site, treatment intent, presence of prior cytotoxic treatment, neuroendocrine differentiation, median culture time or patient survival. The prevalence of p53 mutations in cell lines with ras mutations did not differ from that in cell lines without ras mutations. However, p53 gene mutations in NSCLC cell lines with ras mutations tended to cluster in exon 8, suggesting the presence of a functional domain of the p53 gene relating to interaction with the ras gene. We conclude that p53 and ras mutations are frequent and apparently independent genetic alterations which play different roles in the pathogenesis, progression and prognosis of NSCLC.     

The abnormalities in the p53/p21WAF1 pathway have a significant role in the pathogenesis and progression of gastrointestinal stromal tumors

To elucidate the prognostic role and relationship of the p53/p21/PCNA pathway in gastrointestinal stromal tumors (GISTs), a total of 167 resected gastric and small intestinal CD117-positive stromal tumor specimens were collected from January 1987 to December 2000. Immunohistochemical studies were performed on the paraffin sections with antibodies of p53, p21/WAF1, proliferating cell nuclear antigen (PCNA) and Ki-67. The immunoreactivity of four markers was recorded by labeling index (LI, %) for clinicopathologic and survival correlation. The labeling index was 0-83% for p53, 0-81% for p21/WAF1, 0-33% for Ki-67 and 12-92% for PCNA. Completely negative immunostaining (LI<1%) was found in 54.5% of p53, 25.8% of p21/WAF1 and 44.3% of Ki-67. The LI of four markers strongly correlate with each other (p<0.05). Furthermore, the LI of all four markers positively correlate to microscopic tumor mitotic counts (p<0.05). Only the LI of p53 and PCNA positively correlate to tumor sizes. Tumors with non-spindle cell type (versus spindle cell) and high cellular pleomorphism (versus low) exhibited a higher p53, p21/WAF1 and PCNA LI (p<0.05). Increased NIH risk significantly correlates to increased p53, PCNA and Ki-67 (p<0.05) LI. Survival analysis indicated that a large tumor size (> or =5 cm, p=0.003), increased tumor mitosis (> or =5/50 HPF, p<0.001), high NIH risk (p<0.001), non-spindle cell type (p=0.024), high p53 LI (p<0.001), high p21/WAF1 LI (p=0.007), high Ki-67 LI (p<0.001) and high PCNA LI (p<0.001) were prognostic factors for poor disease-free survival. Independent factors are tumor size, NIH risk, p53 and p21/WAF1 LI. We demonstrated the first evidence of the linear relationship and prognostic role of the p53/p21/PCNA pathway in gastrointestinal stromal tumors. Abnormalities of the p53/p21WAF1 pathway lead to increased proliferating states, thereby triggering the progression of GISTs.     

Expression of ras Rb1 and p53 proteins in human breast cancer.

The ras, Rb and p53 genes have been implicated in the development of human breast cancer. Qualitative or quantitative changes in the expression of the ras p21 may lead to cell transformation, and this has been previously demonstrated in breast cancer. Both the retinoblastoma protein (Rb1) and the p53 gene product appear to function as negative regulators of cell division. We have investigated the expression of ras p21, Rb1 and p53 proteins in human breast cancer patients immunohistochemically, and correlated the results with a range of clinical and pathological parameters. Ras p21 expression was elevated in 65 per cent and p53 in 23 per cent of cases. Rb1 was expressed in 58 per cent of breast cancer tissues and in 75 per cent of normal tissue. Only four patients were found to have loss of Rb1 expression and also overexpression of both p53 and ras gene products. No correlations were found between the expression of these three genes and menopausal status, histological types or tumour grade. However, a correlation was found between Rb1 loss of expression and tumour diameter (greater than 2 cms), and no lymph node metastasis. Also, a significantly higher number of p53 staining specimens were found to be overexpressing the ras gene. These results suggest that all three oncogenes are most likely involved in the development of breast cancer but that their role is complex.     

RAS AND p53 EXPRESSION IN HUMAN THYROID CARCINOMA

Both benign and malignant nodular disorder of the thyroid gland may give rise to similar symptomatology. Even though clinical background and pathologic criteria may predict prognosis, there are still patients without these adverse prognosis indicators who develop subsequent local invasion or distant metastasis after surgical intervention and eventually succumb to the disease.[1] In recent years it has become apparent that malignant transformation is a result of the accumulation of genetic abno…     

Disruption of p53-p21/WAF1 cell cycle pathway contributes to progression and worse clinical outcome of hepatocellular carcinoma

p53-p21/WAF1 cell cycle pathway plays an important role in growth control, and the inappropriate deregulation of this pathway has been implicated in carcinogenesis. Although the role of p53 in hepatocellular carcinoma (HCC) has been suggested, its exact molecular mechanism in relation to its down-stream gene p21/WAF1 remains unclear. To investigate the relationship between the expression of p53 and p21/WAF1 and the possible roles of the 2 proteins in HCC, we examined the intracellular expression of p53, p21/WAF1 and PCNA immunohistochemically, together with apoptosis by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling assay in 35 clinical tissue specimens. The correlation between the clinicopathologic parameters and the intracellular gene expression were analyzed. The results showed that p53 over-expression is a reliable marker for mutational modulation of p53 function. p53 was negatively correlated with p21/WAF1 in hepatitis B virus-related HCC (p=0.024, r=-0.432). Patients with a high p53 expression had a significantly higher Edmondson grading (12/21 vs 13/14, p=0.024) and larger tumor size (10 vs 6 cm, p=0.029). Patients with higher p53 expression had shorter disease-free survival (4 vs 19 months, p=0.0131) and overall survival (11 vs 42 months, p=0.0031). Intracellular expression of p21/WAF1 was positively correlated to proliferating cell nuclear antigen (p=0.001, r=0.776) and apoptosis (p=0.003, r=0.639). Our findings suggest that disruption of p53-p21/WAF1 cell cycle pathways contributes to tumor progression and worse clinical outcome of HCC.