Serum p53 antibodies in patients with lung cancer: correlation with clinicopathologic features and smoking

Abnormalities of p53 gene can lead to the production of p53 antibodies (p53-Abs) in the serum of cancer patients. This study was designed to investigate the prevalence of p53-Abs in 133 lung cancer patients and the distribution of these antibodies to clinicopathologic features and smoking status. Twenty five (18.8%) lung cancer patients were found to have p53-Abs. The presence of p53-Abs did not correlate with sex or age but showed frequent association with tumors of squamous cell carcinoma (31%) in comparison with adenocarcinoma (13.6%) (P=0.052). There was a statistically significant difference in the incidence of p53-Abs between early disease group (stage I-II) and the advanced group (stage III-IV) (P=0.036), however, there was no relationship between the presence of p53-Abs and overall survival. Interestingly, the frequent of p53-Abs was higher in smokers (27.1%) than in non-smokers (13.6%), though the difference was of borderline of statistical significance (P=0.061). These findings suggested that p53-Abs could be a potential biomarker for the study of individual with lung cancer.     

Serum antibodies against p53 in relation to cancer risk and prognosis in breast cancer: a population-based epidemiological study

To perform an epidemiological evaluation of the predictive value of p53 autoantibodies in breast cancer, we measured antibodies against p53 in serum samples from 165 breast cancer patients in comparison with serum samples from 330 healthy controls, selected from the same population as the cases and matched for age, sex and specimen storage time. Median age of patients was 51 years (range 25-64 years). Presence of serum p53 autoantibodies was analysed by enzyme-linked immunosorbent assay (ELISA) and confirmed by Western blotting. The lower ELISA reactivities were similar for cases and controls, but presence of high-level reactivity was more common among cases than among controls [odds ratio (OR) 9.03, 95% confidence interval (CI) 2.40-50.43]. Presence of Western blot-detected p53 autoantibodies had a very similar association (OR 10.8, CI 3.0-59.4). Among the cases, we also studied whether there was any correlation between level of anti-p53 antibodies and stage of the disease or survival. There was no significant correlation between presence of antibodies and stage of the disease. There was a significant negative correlation between presence of p53 antibodies and survival (P = 0.003). A stepwise multivariate Cox regression analysis showed that T-stage, age and presence of anti-p53 antibodies were significant independent prognostic variables, with a dose-dependent negative effect on survival for all three variables. We conclude that presence of anti-p53 antibodies are of significance both for the risk of having breast cancer and the risk of dying from breast cancer.     

Telomerase activity in relation to p53 status and clinico-pathological parameters in breast cancer

Cell cycle deregulation can occur at different levels in cancer. In human breast cancer it includes overexpression of cyclins D1 and E, down-regulation of cyclin-dependent kinase inhibitors and inactivation of the retinoblastoma and p53 tumor suppressor proteins. Telomerase activity is strongly associated with an immortal phenotype and expression of telomerase is linked to the cell cycle. We have recently demonstrated a connection between specific cell cycle defects within the pRB pathway and levels of telomerase activity in breast cancer. In the present study, 106 tumors were investigated for p53 gene and protein status. By single strand conformation polymorphism (SSCP) analysis, 15% showed mutations within exons 5–8 and by immunohistochemistry (IHC), 29% were p53 positive. Tumors with a telomerase activity above median (i.e., telomerase^high) were significantly associated with p53 protein accumulation (p = 0.004), but not with p53 gene mutations. The strongest telomerase expression was found in tumors with p53 protein accumulation. Morphologic grade, estrogen and progesterone receptor expression differed significantly between the telomerase^high and telomerase^low groups (p < 0.0001, p = 0.016 and p = 0.046, respectively), but no difference was observed for stage or nodal status. Telomerase^high tumors were significantly associated with a poor prognosis for node-negative (N0) patients (p = 0.008), but not for node-positive (N⁺) patients, whereas the opposite was demonstrated for tumors with p53 accumulation. The survival data indicated that telomerase expression has biological importance particularly for N0 tumors, suggesting that telomerase^low tumors constitute a group of “pre-immortalized” tumors with a good prognosis. Int. J. Cancer (Pred. Oncol.) 79:343–348, 1998. © 1998 Wiley-Liss, Inc.     

COX-2和p53在乳腺癌组织中的表达及其相关性

目的:研究乳腺癌中环氧化酶-2(cy-clooxygenase-2,COX-2)和p53的表达及其相关性。方法:采用免疫组织化学法检测48例乳腺癌组织中COX-2与p53的表达情况,分析其相互间关系及与乳腺癌临床病理学特征间的联系。结果:乳腺癌中COX-2表达阳性率60·4%(29/48)。COX-2高表达与肿块>2cm、淋巴结转移阳性、ER阴性、PR阴性显著相关,而与患者年龄、肿瘤病理类型及肿瘤分期无明显关系。在p53阳性的乳腺癌患者中COX-2表达阳性率82·6%(19/23),而p53阴性的乳腺癌患者中,COX-2表达阳性率只有40·0%(10/25),两者相比有统计学意义,P=0·006。结论:COX-2在乳腺癌中高水平表达且与p53密切相关,提示乳腺癌中COX-2与p53存在互相调控机制,共同促进肿瘤的发生和发展。     

乳腺癌改良根治术患者生物学指标的临床研究

目的：探讨乳腺癌改良根治术患者乳腺癌组织中ＥＲ、ＰＲ、Ｃ-ｅｒｂＢ-２、Ｃｌｕｓｔｅｒｉｎ、ｐ-ｇｐ表达之间的相关性。方法：免疫组化法检测４８例乳腺癌改良根治术患者乳腺癌组织中ＥＲ、ＰＲ、Ｃ-ｅｒｂＢ-２、Ｃｌｕｓｔｅｒｉｎ、ｐ-ｇｐ的表达,应用数据挖掘技术对乳腺癌５种生物学指标进行分析。结果：４８例乳腺癌改良根治术患者乳腺癌组织中,ＥＲ阳性率为４３.８％,ＰＲ３５.４％,Ｃ-ｅｒｂＢ-２６０.４％,Ｃｌｕｓｔｅｒｉｎ６２.５％,ｐ-ｇｐ４１.７％,Ⅱ期ｐ-ｇｐ、Ｃｌｕｓｔｅｒｉｎ表达的阳性率低于ⅢＡ期（Ｐ＜０.０５）。指标聚类分析提示,ＥＲ、ＰＲ和Ｃ-ｅｒｂＢ-２之间具有较强的正相关性,它们与ｐ-ｇｐ、Ｃｌｕｓｔｅｒｉｎ分别呈较弱的负相关性,而ｐ-ｇｐ与Ｃｌｕｓｔｅｒｉｎ之间呈较强的正相关性。二阶段聚类分析结果提示,４８例乳腺癌患者的５个分子标记物情况可分为３类：第１类：Ｃｌｕｓｔｅｒｉｎ（－）、ｐ-ｇｐ（－）、Ｃ-ｅｒｂＢ-２（＋）和ＥＲ（＋）;第２类：Ｃｌｕｓｔｅｒｉｎ（＋）、ｐ-ｇｐ（＋）和Ｃ-ｅｒｂＢ-２（＋）;第３类：Ｃ-ｅｒｂＢ-２（－）、ＥＲ（－）和ＰＲ（－）。结论：Ｃｌｕｓｔｅｒｉｎ和ｐ-ｇｐ在乳腺癌中的表达与ＥＲ、ＰＲ和Ｃ-ｅｒｂＢ-２的表达存在明显的差异且相关性弱,有可能是乳腺癌治疗的新靶点。     

p53 mutations in breast cancer.

We have identified and analyzed 41 mutations in p53 in sporadic breast tumors from 136 unselected breast cancer patients and estimate that approximately 40% of such tumors contain p53 mutations. The frequency of G-T transversions and the incidence of guanosine mutations in the nontranscribed strand of the p53 gene were found to be higher than expected, and we suggest, therefore, that exogenous carcinogens have an etiological role in sporadic breast cancers. Mutations were recorded in 44 codons of the p53 gene, with no obvious mutational hot-spots, although mutations at codons 175, 194, 273, and 280 accounted for 25% of the changes. One germ-line mutation was found in 136 patients and so we conclude that constitutional mutation of p53 may be an uncommon etiological factor in breast cancer.     

Serum p53 antibodies in small cell lung cancer: the lack of prognostic relevance

Prognostic relevance of serum p53 antibodies was assessed in 96 patients with microscopically proven small cell lung cancer (SCLC). The study group included 67 males and 29 females; mean age 58 years; range 35--86 years; 60 with limited disease (LD), and 36 with extensive disease (ED). The control group consisted of 41 patients with non-malignant diseases. The presence of p53 antibodies was assayed by the immunoenzymatic method (P53 ELISA kit, PharmaCell, France). Antibodies were present in 26 SCLC cases (27%); 15 (25%) in LD and 11 (31%) in ED. Antibodies were also found in one out of 41 control subjects (2%). There was no correlation between the level of antibodies and clinical characteristics of SCLC patients including age, gender and extent of disease. The median follow-up for the entire group was 30 months (range: 11--39 months). By the time of analysis, 78 patients (82%) had deceased. Median survival in SCLC patients with and without antibodies was 42 and 39 weeks, respectively (log rank, P=0.81). These results indicate the lack of clinical relevance of serum p53 antibodies in SCLC.     

Detection of serum anti-p53 antibodies from patients with ovarian cancer in China: correlation to clinical parameters

Background: Ovarian cancer has been one of the most common malignant tumor among women in China. The aims of this research were to increase the detection efficiency of anti-p53 antibodies in the sera from patients with ovarian cancer and to assist the diagnosis for patients with ovarian cancer. Methods: The hybrid phage displaying the immunodominant epitope SQAMDDLMLS in p53 N-terminal region was constructed and the fusion protein was prepared and purified. Ninety-two nonselected Chinese women with ovarian cancer were involved in this study. Tumor p53 overexpression was assessed by immunohistochemical analysis of tissue sections. Serum antibodies to p53 were also detected by enzyme-linked immunosorbent assay (ELISA) using recombinant human wild-type p53 protein and the hybrid phage as the coating antigen respectively. Furthermore, the correlations between the anti-p53 antibodies and clinicopathological parameters were analyzed. Results: The positive rate of anti-p53 antibodies in the patients with ovarian cancer was increased (39.1%, 36/92) through the combination of the two ELISA methods compared with each method. The anti-p53 antibodies were not associated with the clinicopathologic parameters, while there was a significant correlation between the presence of p53 Abs and tissue overexpression of p53 in ovarian cancer. Conclusion: These preliminary results demonstrate that the combination of the two ELISA methods increased the positive rate of anti-p53 antibodies in patients with ovarian cancer and provided a useful marker to complement routine clinical diagnosis for patients with ovarian cancer.     

Expression of p43 in breast cancer tissue, correlation with prognostic parameters.

Placental isoferritin (PLF) and its unique superheavy chain p43 have been recently described as being synthesized by breast cancer cell lines but not by normal breast epithelial cells. Since previous reports have demonstrated a correlation between the content of 'normal' ferritin in breast cancer tissue and the degree of differentiation and prognosis, we have determined p43 in the cytosol of 122 breast cancer samples by use of the new monoclonal antibody CM-H-9. The synthesis of p43 showed a significantly negative correlation with tumor size (P = 0.0001), histologic grading (P = 0.0038), nuclear pleomorphism (P = 0.0019), rate of mitosis (P = 0.0002), lymphocytic reaction (P = 0.0001) and a significantly direct correlation with estrogen receptor status (P = 0.0009). Although patients with a higher p43 content showed a trend for a better outcome (median follow-up: 61.4 months), an independent influence of the cytosolic p43 content on survival could not be confirmed by a multiple Cox model. Therefore it seems that p43's prognostic impact is linked to the highly significant correlation with features of differentiation although a statistical bias in the Cox model due to the limited number of patients must also be taken into account. On the other hand, the significant correlation of p43 expression with factors for good prognosis was striking and consistent and warrants further research of this tumor product.     

Serum anti-p53 antibodies in patients with lung cancer

Antibodies against the p53 protein are produced by some cancer patients. In some tumour entities, the presence of p53 autoantibodies have been linked to poorer survival. This study was designed to assess the prevalence and prognostic implications of p53 autoantibodies in patients with lung cancer. Serum samples of 180 patients were tested for antibodies against p53 protein using an ELISA. We studied 134 patients with primary lung cancer [histology: small cell lung carcinoma (SCLC) n=35; non-small cell lung carcinoma (NSCLC) n=99]. The control group consisted of 46 patients without lung cancer. In 17/134 (12.6%) of the cancer patients, p53 autoantibodies were detected (4/35 SCLC, 13/99 NSCLC). Most of the positive results were found in advanced stages of NSCLC (stage I-IIIA: 1/34; stage IIIB/IV: 12/65). One of the 46 control patients tested positive. Statistical analysis of survival shows no correlation with p53 antibody status in SCLC, but a significant correlation with shorter survival in NSCLC (p=0.01). After correction for stage of disease this correlation remains significant (stage IIIB/IV: p=0.02). In our series, the presence of anti-p53 autoantibodies is almost exclusively linked to the presence of malignant disease. Prognosis for patients with NSCLC, but not SCLC seems to be linked to the p53 autoantibody status.     

Combining intracellular antibodies to restore function of mutated p53 in cancer

TP53 is a tumor suppressor gene that is mutated in 50% of cancers, and its function is tightly regulated by the E3 ligase, Mdm2. Both p53 and Mdm2 are localized in the cell nucleus, a site that is impervious to therapeutic regulation by most antibodies. We identified a cell‐penetrating lupus monoclonal anti‐DNA antibody, mAb 3E10, that targets the nucleus, and we engineered mAb 3E10 to function as an intranuclear transport system to deliver therapeutic antibodies into the nucleus as bispecific single chain Fv (scFv) fragments. Bispecific scFvs composed of 3E10 include PAb421 (3E10‐PAb421) that binds p53 and restores the function of mutated p53, and 3G5 (3E10‐3G5) that binds Mdm2 and prevents destruction of p53 by Mdm2. We documented the therapeutic efficacy of these bispecific scFvs separately in previous studies. In this study, we show that combination therapy with 3E10‐PAb421 and 3E10‐3G5 augments growth inhibition of cells with p53 mutations compared to the effect of either antibody alone. By contrast, no enhanced response was observed in cells with wild‐type p53 or in cells homozygous null for p53.     

The correlation of extranodal invasion with other prognostic parameters in lymph node positive breast cancer

BACKGROUND: The presence of extranodal invasion (ENI) in the metastatic lymph nodes is reported to increase the risk of locoregional recurrence while shortening disease-free and overall survival in patients with breast cancer. In this study the relationship between ENI and other prognostic parameters and survival is investigated. METHODS: Of 650 patients with breast cancer who were treated in Ankara Oncology Teaching and Research Hospital from 1996 to 2003, 368 (56.6%) had lymph node metastasis. The patients with axillary metastasis were separated into two groups as with and without invasion to lymph node capsule and the surrounding adipose tissue. Clinicopathologic features were analyzed by univariate and multivariate logistic regression. RESULTS: Of 368 patients with axillary metastasis, 135 (36.7%) had ENI. Based on multivariate analysis; the number of metastatic lymph nodes, lymphatic invasion, and tumor necrosis were found to be related with ENI. In the group with ENI, 5-year overall survival rate was 74.8%, compared to 82.3% for patients without ENI which was significantly lower (P = 0.04). CONCLUSIONS: In lymph node positive breast cancer with presence of ENI, adverse prognostic parameters are more frequently encountered and has a worse overall survival compared to group without ENI.     

p53 polymorphisms and haplotypes in breast cancer

Three polymorphisms in the human tumor suppressor gene p53 (BstUIand MspI RFLPs in exon 4 and intron 6 respectively and a 16bp duplication in intron 3) and their haplotype combinationswere studied in patients with breast cancer and controls. Asignificant increase in the codon 72 BstUI A1 (pro) allele frequency(p= 0.016) and of individuals carrying the pro allele (pro/proand pro/arg) (OR,1.47; p = 0.014; 95% CI, 1.08–2.00) wasobserved in breast cancer. This increase wasmost pronouncedin highly differentiated breast cancer. Significant associationswere found only in BstUIand haplotypes containing this polymorphism,which indicates that the codon 72 pro allele may be functionallyinvolved in low malignancy breast cancer. The distributionsof genotypic combinations in breast cancer patients and controlswere significantly different (p = 0.005). Two BstUI–16bp-MspI combinations were significantly overrepresented; 2–1,1–1, 2–2 (OR, 1.61; 95% CI, 1.13–2.30) and1–1, 2–1, 2–1 (OR, 2.94; 95% CI, 1.37–6.27).     

Survivin在乳腺癌中的表达及其与p53的关系

目的:观察Survivin在乳腺癌及癌旁组织中的表达情况,探讨其与突变型p53基因表达的相关性。方法:用免疫组化SP法检测60例乳腺癌组织,60例癌旁乳腺正常组织中Survivin和p53的表达情况,分析其与腋窝淋巴结转移之间的关系。结果:60例乳腺癌组织中Survivin蛋白的阳性表达率为78.33%,明显高于癌旁正常组织(5.00%)(P<0.05),Survivin在p53阳性的乳腺癌组织中的表达率明显高于p53阴性组,Survivin的表达与乳腺癌的病理分级和淋巴结转移状况无显著相关。结论:乳腺癌组织中Survivin表达上调与乳腺癌的发生与发展密切相关,其表达与p53基因突变显著相关,两者可能协同作用参与乳腺癌的发生与发展。     

p53 allele losses, mutations and expression in breast cancer and their relationship to clinico-pathological parameters.

The p53 locus on the short arm of chromosome 17 at 17p 13.1 was examined for loss of heterozygosity, mutation, mRNA and protein expression in 60 primary breast cancers. Allele loss around the p53 locus was detected in 19/45 informative tumours (42%). p53 mutations in the evolutionarily conserved exons 5 to 9 were detected in 17/60 (28%) by amplification mismatch and confirmed by direct DNA sequencing. p53 mRNA expression was detected by Northern blot in 36/59 (61%) of tumours, and p53 protein expression using antibody 1801 on frozen-tissue sections in 13/44 of the tumours examined. p53 mutation was significantly associated with oestrogen-receptor-poor tumours (p less than 0.01) and hence with poor prognosis, but not with other clinical or pathological parameters. There was no statistical correlation between loss of heterozygosity around the p53 locus at 17p13.1 and p53 mutation. Furthermore, p53 mutation was not associated with p53 expression detected by immunohistochemical staining with antibody 1801 or as p53 mRNA. In addition, events on 17p (allele losses, p53 mutation, p53 expression) were independent of c-erbB-2 expression. In breast cancer, by contrast with colorectal, lung and ovarian cancer, there appears to be no clear association between p53 DNA abnormalities and p53 expression.     

Detection of antibodies against the cellular protein p53 in sera from patients with breast cancer

Antibodies reacting with the host protein p53 were found in the sera of patients with primary or secondary carcinoma of the breast. Fourteen out of the 155 sera from breast cancer patients tested were positive for anti-p53 antibodies (9%) and no positives were detected among 164 control sera from normal women tested. The locations of the first metastasis in patients with positive sera were unusual, with more lung metastases and fewer bone metastases than expected. The detection of anti-p53 antibodies indicates that p53 is altered in amount, type or presentation in breast tumors so that it becomes immunogenic.     

Cigarette smoking and other risk factors in relation to p53 expression in breast cancer among young women.

p53 mutations may be a fingerprint for cigarette smoking and other environmental carcinogens, including breast carcinogens. This study was undertaken to explore whether p53 mutations are associated with environmental or other suspected or established risk factors for breast cancer. p53 protein detection by immunohistochemistry (which is more easily quantified in large epidemiological studies than are mutations, and are highly correlated with them) was determined for 378 patients from a case-control study of breast cancer. In this population-based sample of women under the age of 45 years, 44.4% (168/378) of the cases had p53 protein detected by immunohistochemistry (p53+). Polytomous logistic regression was used to calculate the odds ratios (ORs) for p53+ and p53- breast cancer, as compared with the controls, in relation to cigarette smoking and other factors. The ratio of the ORs was used as an indicator of heterogeneity in risk for p53+ versus p53- cancer. The ratio of the ORs in a multivariate model was substantially elevated among women with a greater than high school education [2.39; 95% confidence interval (CI), 1.43-4.00], current cigarette smokers (1.96; 95% CI, 1.10-3.52), and users of electric blankets, water beds, or mattresses (1.78; 95% CI, 1.11-2.86). Nonsignificant heterogeneity was noted for family history of breast cancer and ethnicity but not for other known or suspected risk factors. Coupled with the strong biological plausibility of the association, our data support the hypothesis that in breast cancer, as with other tumors, p53 protein immunohistochemical detection may be associated with exposure to environmental carcinogens such as cigarette smoking.     

Serum p53 antibodies in patients with oral lesions: Correlation with p53/HSP70 complexes

The functional significance of alterations in expression of tumour-suppressor gene p53 and 70-kDa heat-shock protein (HSP70) in eliciting p53-specific humoral response in oral-cancer patients is not yet known. In this study, p53 auto-antibodies were analyzed in sera from oral-cancer patients by immunoblotting and results were correlated with clinicopathological features of the patients as well as with the levels of HSP70, p53 and p53-HSP70 complexes in matched patients' tumour tissue, for determining their diagnostic/prognostic significance and relationship to survival. Circulating anti-p53 antibodies were observed in 7 of 30 cancer patients and 3 of 25 patients with pre-malignant lesions. Over-expression of p53 protein in matched oral lesions was observed in 22 of these 30 cancer patients and 14 of 25 patients with dysplastic lesions. No detectable levels of p53 protein or anti-p53 antibodies were observed in normal subjects (15 cases). Elevated levels of HSP70 were observed in 23 of these 30 oral tumours and 17 of 25 dysplastic lesions. All the anti-p53-antibody-seropositive cases showed elevated levels of p53 and HSP70 proteins, as well as formation of p53-HSP70 complexes, in matched dysplastic or malignant lesions, suggesting that these molecular alterations may be early events in oral tumorigenesis and are implicated in eliciting p53-specific humoral immune response in these patients. Anti-p53-antibody-seropositive cases showed poor prognosis and significantly decreased overall disease-free survival in comparison with the seronegative cases. Detection of circulating anti-p53 antibodies may serve as a useful non-invasive marker for identifying oral tumours having poor prognosis. Int. J. Cancer 74:609–613, 1997.© 1997 Wiley-Liss, Inc.