原癌基因HER2/neu表达产物p185蛋白免疫原性研究

p185蛋白是由原癌基因HER2/neu编码的跨膜糖蛋白,在许多肿瘤患者体内高表达,与疾病的预后密切相关,表达增高预后较差.已从肿瘤患者体内分离出识别p185蛋白及多肽CTL和CD4~ 的T细胞,也证实患者血清中存在抗p185抗体,p185作为肿瘤抗原已引起广泛关注.为进一步研究将该蛋白作为瘤苗应用于临床可行性,我们用表达p185蛋白的HER2/neu转基因3T3-HER2/neu细胞裂解物免疫BABL/c小鼠,经3次免疫后,免疫鼠脾细胞对表达p185蛋白3T3HER2/neu细胞呈现较强的增殖反应(平均刺激指数分别为SI=3.15±1.88),而对未转染HER2/neul基因的3T3细胞的应答较弱(SI=1.14±0.97).并检测到1号免疫鼠脾细胞对3T3-HER2/neu细胞有特异杀伤.免疫鼠血清中出现高水平的抗P185抗体(OD均值=1.02±0.16),较正常对照组(OD     

p185蛋白及其抗体与乳腺癌的关系

p185蛋白是癌基因C-erbB-2(又称neu或HER-2)编码的具有酪氨酸激酶活性的跨膜受体蛋白,激活后参与调控细胞的有丝分裂、增殖和分化[1-2].Ravdin等[3]认为约有20%～30%的乳腺癌和卵巢癌的p185蛋白呈高表达,且p185蛋白的高表达对乳腺癌,其是淋巴结阳性亚群有重要的预后判断价值.Leitzei等[4]发现 p185蛋白还有一种分泌到血清中的形式,种分泌形式在浸润性较高的乳腺癌患者中过表达,作为肿瘤诊断和监测的指标.Hynes等[5]发现p185蛋白的高表达能使患者对化疗不敏感,此研究p185蛋白及其抗体与乳腺癌的关系具有重要意义.     

ELISA法检测血清中P-185的临床应用

目的:建立一种快速简单,用于定量检测血清中P-185含量的双夹心ELISA检测法,并比较研究正常人和恶性肿瘤患者血清中P-185含量及其临床意义.方法:采用ELISA双夹心法,检测193例正常人、133例恶性肿瘤患者和34例肝硬化患者血清.结果:正常人外周血中可以检测到低水平P-185;恶性肿瘤患者和肝硬化患者血清中P-185表达水平显著高于正常人(P＜0.05).结论:ELISA法检测血清中P-185表达是可行的,P-185过度表达可以作为恶性肿瘤早期诊断标志和判断预后指标;对P-185表达进行动态监测,可以判断预后和指导临床治疗.     

血清p53抗体与肺癌生物学行为和化疗疗效的关系

目的通过对血清p53抗体与肺癌生物学行为和化疗疗效关系的研究,探讨血清p53抗体在肺癌中的临床意义。方法对57例初步诊断为肺癌的患者应用ELISA法检测血清p53抗体效价,经酶联免疫检测仪测吸光值E450,计算抗体指数,进行统计学分析。结果57例中阳性25例,阳性率为43.9%。血清p53抗体水平与肺癌的一般临床特征,如性别、吸烟指数无关,而与肺癌的病理类型、分期、分化和淋巴结转移有关。结论p53抗体的产生可能是肺癌发生的早期指征,是肺癌的不良预后因子,化疗前测定血清p53抗体可以预测化疗疗效。     

一种检测非小细胞肺癌患者血清中p53自身抗体的新方法

背景与目的 多肽阵列是近年来发展起来的一种研究分子之间相互作用的方法.本研究拟建立多肽阵列检测非小细胞肺癌患者血清中p53自身抗体的方法,筛选p53自身抗体识别的抗原决定簇序列.方法 利用点合成技术(SPOT synthesis technique)合成覆盖p53多肽链全部序列的多肽阵列;分别用ELISA和多肽阵列检测血清中的p53自身抗体,确定p53自身抗体识别的抗原决定簇序列.结果 将包含p53多肽链全部序列的重叠肽段合成到硝酸纤维素膜上.多肽阵列检测30例非小细胞肺癌患者血清中的p53自身抗体.有7例为阳性;30例健康对照者均为阴性,与ELISA检测结果相同.P53自身抗体识别p53多肽链上某些共同的抗原表位.结论 多肽阵列不仅可以用来检测肺癌患者血清中的自身抗体,还可以筛选这些自身抗体识别的抗原表位,为下一步研究自身抗体与非小细胞肺癌的早期诊断和预后的关系奠定基础.     

ELISA法检测血清中P-185的临床应用

目的 :建立一种快速简单 ,用于定量检测血清中P -185含量的双夹心ELISA检测法 ,并比较研究正常人和恶性肿瘤患者血清中P -185含量及其临床意义。方法 :采用ELISA双夹心法 ,检测 193例正常人、133例恶性肿瘤患者和 34例肝硬化患者血清。结果 :正常人外周血中可以检测到低水平P 185 ;恶性肿瘤患者和肝硬化患者血清中P -185表达水平显著高于正常人 (P <0 .0 5 )。结论 :ELISA法检测血清中P -185表达是可行的 ,P- 185过度表达可以作为恶性肿瘤早期诊断标志和判断预后指标 ;对P- 185表达进行动态监测 ,可以判断预后和指导临床治疗。     

P185蛋白表达与血清CA153联合监测在乳腺癌治疗中的意义

目的 探讨乳腺癌组织中P185蛋白表达与血清糖蛋白抗原153(CA153)的相关性及联合监测的临床意义.方法 通过免疫组化法和放射免疫法监测2005年6月至2007年5月间110例乳腺癌P185表达情况及血清CA153的水平,分析P185蛋白、CA153与乳腺癌临床特征之间的关系.结果 乳腺癌患者治疗前血清CA153的水平明显高于治疗后的水平(P<0.01),乳腺癌Ⅲ、Ⅳ期患者的血清CA153的水平明显高于Ⅰ、Ⅱ期患者的水平(P<0.01),P185蛋白高表达患者的血清CA153水平高于低表达患者(P<0.05).结论 联合监测P185蛋白与血清CA153可以用于乳腺癌的预后判断,指导辅助治疗方案的制定.     

血清COX-2在乳腺癌中的表达及与临床病理特征关系的研究

目的 利用酶联免疫吸附试验(ELISA)方法 检测乳腺癌患者血清环氧化酶-2(cyclooxygenase-2,COX-2)并探讨其与临床病理特征之间的关系.方法 随机抽取2009年9月至2010年9月柳州工人医院普通外科收治的乳腺癌患者65例,乳腺良性病变患者30例,正常人25例.采用ELISA法检测3组入选者血清COX-2水平,并作统计学分析,探讨COX-2与各项临床病理特征之间的关系.结果 血清COX-2在乳腺癌中均有表达.在65例乳腺癌中,血COX-2含量临床分期Ⅲ+Ⅳ期组比0+Ⅰ+Ⅱ期组高;有腋窝淋巴结转移者比无腋窝淋巴结转移者高;ER、PR阴性组较阳性组高;p21阳性组较阴性组高.结论 ①COX-2表达与乳腺癌发生、发展有关;②COX-2含量与患者TNM分期、腋淋巴结转移情况、ER、PR、p21有关.检测COX-2有助于评价乳腺癌患者的临床及预后.     

女性乳腺癌患者前列腺特异性抗原检测的临床意义

本文应用双抗体夹心ELISA法对24例女性乳腺癌患者测定其血清PSA总量,并作雌激素受体/孕激素受体(ER/PR)免疫组化检测,旨在探讨其在乳腺癌诊断和预后分析中的临床意义.     

重组小鼠Izumo的表达及其特异性抗体对小鼠体外精卵融合的影响

背景与目的:精子膜蛋白Izumo在精卵融合中起关键作用.本研究旨在表达和纯化重组小鼠Izumo蛋白(mIzumo),探讨重组mIzumo在小鼠体内的免疫原性及其特异性抗体对小鼠体外精卵融合的影响.材料与方法:将编码mIzumo蛋白的cDNA序列亚克隆到pET28a(+)原核表达载体上,构建pET28a(+)-mIzumo重组质粒.在BL21(DE3)大肠杆菌中表达重组6His-mIzumo融合蛋白,并通过Ni2+亲和层析纯化.使用福氏佐剂乳化纯化的6His-mIzumo,分别免疫雌性和雄性C57BL/6小鼠,并采集免疫前和免疫后血清.用Western Blot和ELISA检测血清中抗6His-mIzumoIgG抗体的特异性和滴度.采用IVF和精卵融合试验检测血清中抗6His-mIzumo抗体对小鼠精卵融合的影响.结果:纯化的6His-mIzumo在SDS-PAGE凝胶上显示为约60 kD的单一条带.免疫了重组蛋白的雌性和雄性小鼠都产生了抗6His-mIzumoIgG抗体.抗6His-mIzumoIgG抗体与重组蛋白,小鼠睾丸、附睾及精子膜蛋白存在交叉反应,免疫印迹显示为约60kD的特异性条带.ELISA结果表明,免疫6His-mIzumo后至少6周之内,血清中的抗6His-mIzumoIgG抗体维持在最高水平.免疫后血清处理组的小鼠精子与去透明带卵母细胞发生胞膜融合的能力明显低于免疫前血清处理组(P＜0.01).结论:纯化的6His-mIzumo融合蛋白能够诱发雌性和雄性小鼠产生可阻断精卵融合发生的特异性血清抗体.因此,6His-mIzumo可作为同种异体抗原用于免疫避孕候选疫苗的研究.     

Detection of serum p53 protein in patients with different gastrointestinal cancers

Overexpression of p53 has been found in many types of human malignancy. The present study aimed to detect preoperative serum p53 among 158 patients with different gastrointestinal cancers using ELISA technique based on mouse anti-p53 DO-7 monoclonal antibody and anti-p53 rabbit polyclonal antibody. A single band of 53kDa was detected in nuclear protein tissue extracts of selected cancer patients and in 96% of the corresponding sera using Western blot assay. The ELISA technique revealed that the serum p53 was detected in 100% of patients with cholangiocarcinoma, 76% of pancreatic carcinoma, 75% of hepatocellular carcinoma, 70% of colon cancer, 60% of esophagus carcinoma, and 35% of gastric carcinoma. The serum p53 concentrations of the positive patients were highly elevated (P<0.001) compared with healthy individuals. These results suggest that immunodetection of serum p53 could be valuable for post-operative monitoring during follow up in preoperatively positive patients with gastrointestinal cancers.     

Measurement and evaluation of serum anti-p53 antibody levels in patients with lung cancer at its initial presentation: a prospective study.

Anti-p53 antibodies in sera are known to be products of the host immune response to mutated p53 protein, and are present in some patients with various types of cancer. In this study, we measured serum anti-p53 antibody levels in 52 patients with lung cancer and 63 normal volunteers to determine the relationship between anti-p53 antibody level and clinical features of lung cancer patients. Anti-p53 antibody level was measured by an enzyme-linked immunosorbent assay and expressed as an anti-p53 antibody index, defined as the ratio of absorption of serum sample to that of p53-positive serum. The median anti-p53 antibody index was 6.6 for lung cancer patients, and higher than that in normal volunteers (1.7) (P = 0.0000). For lung cancer patients, significant differences in index levels were found by histology (4.3, n = 25, adenocarcinoma vs 8.7, n = 18, squamous cell carcinoma vs 64.8, n = 2, large-cell carcinoma vs 9.8, n = 7, small-cell carcinoma; P = 0.0109). High anti-p53 antibody index levels were observed for both large-cell carcinoma and small-cell carcinoma. When the cut-off level was set at 7.2, determined using the twice 95% specificity level for normal volunteers, the sensitivities of anti-p53 antibodies were 46.1% for all lung cancers, 28.0% for adenocarcinoma, 55.6% for squamous cell carcinoma, 100% for large-cell carcinoma and 71.4% for small-cell carcinoma. However, there were no significant differences in index level by gender, age, smoking index, presence of previous or concomitant cancer or disease stage. Multivariate analysis using a logistic regression model demonstrated that histological type of tumour was a dominant factor associated with elevation of anti-p53 antibody index level (P = 0.0184). These findings suggest that serum anti-p53 antibody index level might be independent of tumour burden and the presence of previous or concomitant cancer in our series of lung cancer patients, but is clearly strongly correlated with tumour histological type.     

Clinical implications of serum anti-p53 antibodies for patients with gastric carcinoma

BACKGROUND: Mutations of p53 can lead to the production of anti-p53 antibodies in sera of cancer patients. Before this study, the value of preoperative serum anti-p53 antibodies in determining the prognoses of patients with gastric carcinoma had yet to be determined. METHODS: The authors used a highly specific enzyme-linked immunosorbent assay (ELISA) kit (Pharma Cell, France) to determine the preoperative presence of serum anti-p53 antibodies in 120 patients with gastric carcinoma. The relation between the positivity of serum anti-p53 antibodies and p53 abnormal staining of gastric carcinoma tissues was examined. Clinicopathologic characteristics and prognoses of these patients were given attention. RESULTS: Anti-p53 antibodies were detected in 19.2%(23 of 120) of these patients with gastric carcinoma. Among those who were positive for anti-p53 antibodies, female patients were predominant, the depth of invasion was greater, and liver metastasis was present, as compared with those who were negative for anti-p53 antibodies. Regarding other factors, there were no differences between those who were positive or negative for anti-p53 antibodies. Gastric carcinoma tissues had a 60.9% (14 of 23) positivity rate of p53 staining with anti-p53 antibodies and a 33.0% (32 of 97) negativity rate, and this difference was statistically significant (P < 0.05). The survival time of patients with anti-p53 antibodies in their sera was shorter than that of subjects with sera negative for anti-p53 antibodies (P < 0.05). The presence of anti-p53 antibodies was not an independent prognostic factor in multivariate analysis. CONCLUSIONS: Serum assay of anti-p53 antibodies is a rapid and readily facilitated test for predicting tumor advancement, depth of invasion, and liver metastasis, and it will show a poorer prognosis for surgically treated patients with gastric carcinoma.     

血清p53抗体水平与肺癌临床指标关系的研究

目的：通过对肺癌患者血清Ｐ５３抗体水平与肺癌临床指标关系的研究，探讨血清ｐ５３抗体在肺癌中的临床意义。方法对６８例肺癌患者应用ＥＬＩＳＡ法检测血清Ｐ５３抗体滴度，经酶联免疫检测仪测吸光值Ｅ４５０，计算抗体指数，进行统计学分析。结果６８例中阳性３２例，阳性率为４７．１％，血清Ｐ５３抗体水平与肺癌的一般临床特征，如性别、分期、吸烟指数、既往治疗均无关，而与肺癌的病理类型和肿块大小有关。结论Ｐ５３抗体的     

Prevalence of antibody against the core protein of hepatitis C virus in patients with hepatocellular carcinoma.

We have cloned the whole structural region of the hepatitis C virus (HCV) genome and transiently expressed the nucleocapsid protein in animal cells. Since the nucleotide sequences of this region of the HCV genome has been shown to be highly conserved among different HCV isolates, the assay detecting the antibody to this expressed protein is useful for studying the pathogenicity of HCV. In this work, we investigated the presence of antibodies to HCV nucleocapsid protein (p22) in patients with hepatocellular carcinoma (HCC) and compared its frequency with that of antibody to HCV non-structural protein (C-100), which is presently applied for blood screening for transfusion and diagnosis for chronic hepatitis C. By a sensitive Western blot analysis, 85 of 102 (83.3%) sera of hepatitis B virus surface antigen (HBsAg)-negative HCC patients were positive for the antibody to p22 (anti-p22), whereas 68 of the same 102 cases (66.7%) were positive for the anti-C100 by ELISA. The prevalence of anti-p22 in 23 HBV carrier HCC patients, 56 patients with non-HCC cancer and 100 healthy blood donors were 4.3, 12.5 and 1.0%, respectively. Thus, high prevalence of anti-p22 in non-B HCC confirmed that HCV infection is closely related to the development of HCC. Furthermore, the anti-p22 assay can detect HCV-infected patients who could not previously be identified as such by the present anti-C100 assay.     

Testing for anti-p53 antibodies increases the diagnostic sensitivity of conventional tumor markers

The aim of this study was to determine whether anti-p53 antibodies are of clinical significance as a serological marker in the diagnosis and monitoring of malignancies. A total of 1874 serum samples from 591 patients with various types of cancer, esophageal, gastric, colorectal, pancreatic, hepatocellular, breast, and urogenital cancer, and 436 control individuals were analyzed by immunoblot for antibodies against p53. The anti-p53 antibody test was correlated with expression of conventional tumor markers, survival and the clinicopathological features of malignant disease. Anti-p53 antibodies were found in 23.4% (138/591) of the sera of patients with malignant disease (range 11.5-34%). The detection of anti-p53 serum antibodies had a specificity of 100% for malignancy (p<0.0001). The overall sensitivity of measuring established tumor markers was 62.9% (372/591). The elevation of conventional tumor markers and the presence of anti-p53 antibodies in the sera of patients with malignant disease turned out to be an independent variable (p<0.05). Combination of established tumor markers with the anti-p53 antibody test led to an increase in diagnostic sensitivity of 8% (49/591) (p<0.01). Thus, the independence of anti-p53 antibodies from established tumor markers allows the serological detection of additional tumor patients. Kaplan-Meier analysis revealed a trend toward a poorer prognosis in hepatocellular carcinoma and breast cancer patients who were anti-p53 serum positive. In conclusion, testing for anti-p53 antibodies can increase the diagnostic sensitivity when used in combination with measurement of conventional tumor markers. This increase is achieved without a parallel decrease in specificity.     

Detection of circulating gastric carcinoma-associated antigen MG7-Ag in human sera using an established single determinant immuno-polymerase chain reaction technique

BACKGROUND: In 1994, a novel sensitive method termed immuno-polymerase chain reaction (PCR) for the detection of the gastric carcinoma-associated antigen MG7-Ag in the gastric carcinoma cell line KATO III was reported. Compared with the enzyme-linked immunoadsorbent assay, the single determinant immuno-PCR technique could allow for as few as 20 cells to be detected and was found to show an approximately 10,000-fold enhancement in sensitivity of the detection limit. The current study clinically evaluated the significance of serum MG7-Ag detection in gastric carcinoma patients. METHODS: The sera of patients were immobilized on wells and a specific DNA molecule, which could be amplified by PCR, was employed as a marker. The biotinylated monoclonal antibody against gastric carcinoma was added to bind the antigen immobilized on the wells. After the biotinylated antibody was bound to the antigen, free avidin was used to attach a biotinylated monoclonal antibody and biotinylated DNA molecule. The biotinylated DNA complexed with antigen-antibody-avidin was amplified by PCR and the PCR products were analyzed by agarose gel electrophoresis. In the current study this method was used to detect circulating MG7-Ag in the sera of patients with gastric carcinoma and other various malignancies. For comparison, carcinoembryonic antigen, CA 50, CA 19-9, and TAG-72 were quantitated by radioimmunoassay and immunoradiometric assay using the relevant commercial kits in the same sera samples from 86 patients with pathologically confirmed gastric carcinoma and 83 patients with relevant benign diseases of the stomach. In addition, the semiquantitative analysis of PCR products among gastric carcinoma patients with or without metastasis was performed to compare the intensity of DNA band amplification. RESULTS: Using the immuno-PCR assay, positive results were obtained in 164 of 198 patients with gastric carcinoma (82.8%). The rates of positivity in other malignancies were 17.4% for esophageal carcinoma (15 of 86 patients), 44.4% for colonic carcinoma (40 of 90 patients), 0% for liver carcinoma (none of 84 patients), 2.2% for ovarian carcinoma (1 of 45 patients), 0% for uterine carcinoma (none of 27 patients), and 6.1% for lung carcinoma (4 of 66 patients). The positive results obtained from those patients with benign diseases were: 7.7% for peptic ulcer (6 of 78 patients), 5.9% for chronic gastritis (7 of 118 patients), 3.3% for chronic colitis (2 of 60 patients), and 0.8% for healthy blood donors (2 of 236 patients). In addition, the semiquantitative analysis of PCR products showed that the intensity of DNA band amplified from the PCR products of those patients with metastasis was much higher than that of patients without metastasis or those with early stage tumors (1.94 +/- 0.03 vs. 1.28 +/- 0.02). In comparative studies of immuno-PCR and commercial assays for tumor-associated antigens the sensitivity of immuno-PCR was 81.4% and pseudopositivity was lower (8.4% vs. 7.2-12.0% with radioimmunoassay or immunoradiometric assay). CONCLUSIONS: The results of the current study demonstrate that introducing PCR into the indirect determination of tumor-associated antigen in the serum can improve the sensitivity of detection greatly. This novel assay also might be used to monitor the circulating amount of tumor-associated antigen after gastrectomy and provide information regarding recurrence or metastasis, as well as for screening elderly patients who have no indications for endoscopy and those with precancerous conditions. The application of immuno-PCR in the serologic diagnosis of carcinoma has significant advantages including ready application in the clinical setting as well as use as a potential screening tool in mass surveys of high risk populations with gastric carcinoma. (c) 2000 American Cancer Society.     

Status of c-erbB-2 in gastric adenocarcinoma: a comparative study of immunohistochemistry, fluorescence in situ hybridization and enzyme-linked immuno-sorbent assay

c-erb-2 amplification and overexpression are currently attracting a great deal of attention because a new adjuvant therapy using an antibody against the c-erbB-2 gene product, trastuzumab (Herceptin; Genentech, Inc., South San Francisco, CA), has proved effective in treating breast cancer with amplification and/or overexpression of c-erbB-2. Aberrations of c-erbB-2 have also been detected in ovarian, endometrial and gastric carcinomas at varied frequencies. Amplification of the c-erbB-2 locus (17q12-q21.32), overexpression of c-erbB-2 protein (p185) and serum levels of soluble c-erbB-2 protein fragments (p105) were examined in gastric cancer patients using fluorescence in situ hybridization (FISH), immunohistochemistry and enzyme-linked immunosorbent assay (ELISA), respectively. Overexpression of c-erbB-2 protein was found in 29 (8.2%) of the 352 gastric carcinomas analyzed. In FISH analysis, all tumors with 3+ immunostaining and 1 of 5 tumors with 2+ staining showed high-level amplification of c-erbB-2. Pre-operative serum p105 was quantified in serum specimens from 129 patients with gastric cancer and 28 patients with benign diseases. There were no significant differences in the serum p105 levels among 11 patients with c-erbB-2-overexpressing carcinomas, 118 patients with c-erbB-2 non-overexpressing carcinomas and 28 controls, although a single case of gastric carcinoma overexpressing c-erbB-2 with extensive liver metastasis had a higher level than the cut-off value. The mechanisms of overexpression of p185 and high-level amplification of c-erbB-2 in gastric adenocarcinomas seem similar to those well-established in breast cancers. Patients having gastric adenocarcinoma with c-erbB-2 amplification are potential candidates for a new adjuvant therapy using humanized monoclonal antibody.     

Complexes between urokinase-type plasminogen activator and its receptor in blood as determined by enzyme-linked immunosorbent assay

Complexes between urokinase-type plasminogen activator (uPA) and its receptor (uPAR) were assessed in plasma and serum from 39 breast cancer patients and from 20 healthy individuals, applying a recently developed enzyme-linked immunosorbent assay (ELISA) for the analysis of these complexes in tumor tissue extracts. The assay is based on a combination of rabbit polyclonal anti-uPA antibodies for catching and a mouse anti-uPAR monoclonal antibody (MAb) for detection. The specificity of the assessment of uPA:uPAR complexes was verified by simultaneous analysis of the individual blood samples in corresponding non-sense ELISA formats, in which either the anti-uPA catching antibody or the anti-uPAR detecting antibody was substituted with an irrelevant antibody. Assessment of native uPA:uPAR complexes was ascertained by demonstrating the absence of any de novo formation of uPA:uPAR complexes in plasma and serum during the sample incubation step in the ELISA, as verified by the use of a peptide antagonist for uPAR. Plasma and serum samples contained almost identical levels of uPA:uPAR complexes. The levels of uPA:uPAR complexes were found to be significantly lower in serum from breast cancer patients compared to the serum of healthy donors, while the levels of (total) uPAR in plasma from breast cancer patients were significantly higher than in plasma from the healthy controls. In addition, the free, uncomplexed uPAR levels, estimated by subtraction of uPA:uPAR complex levels from (total) uPAR levels, were significantly elevated in plasma as well as in serum from breast cancer patients compared to healthy individuals. The uPA:uPAR complex levels were highly comparable to the uPA levels analyzed in the same plasma and serum samples, indicating that most if not all of the uPA present in these samples is complexed with uPAR. Int. J. Cancer 77:236–242, 1998.© 1998 Wiley-Liss, Inc.     

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.