Prognostic value of circulating Bcl-2 and anti-p53 antibodies in patients with breast cancer: A long term follow-up (17.5 years).

BACKGROUND: Apoptosis inhibition is a major tumorigenic factor. Bcl-2 dysregulation and TP53 mutation status, which may correlate with autoantibody generation, contribute to impaired apoptosis. OBJECTIVE: This study aimed to investigate the prognostic value of circulating Bcl-2 and anti-p53 antibodies (p53Abs) in a 17.5-year follow-up of breast cancer patients. We also analyzed the correlations of Bcl-2 and p53Abs with various clinicopathological parameters in order to assess their impact on tumor aggressiveness. METHODS: Serum Bcl-2 and p53Abs levels were analyzed by the enzyme-linked immunosorbent assay (ELISA) in 82 patients with invasive breast cancer and twenty individuals without malignancy. RESULTS: Serum Bcl-2 and p53Abs levels in breast cancer patients were significantly higher than those in controls. Patients with high levels of Bcl-2 (cut-off 200 U/ml) had a poorer prognosis (17.5-year survival) than those with lower Bcl-2 values. In combined analysis the subgroup of patients with elevated p53Abs (cut-off 15 U/ml) and elevated Bcl-2 (cut-offs 124 U/ml and 200 U/ml) had the worse prognosis in 17.5-year survival. In correlation analysis p53Abs and Bcl-2 were associated with unfavorable clinicopathological parameters. CONCLUSIONS: Our results suggest that breast cancer patients with high serum levels of p53Abs and Bcl-2 present an especially unfavorable group in a long follow-up.

Enzyme-Linked Immunosorbent Assay Detection For Malignancy Using Anti-P53 Antibodies

@#Tumour protein 53 (p53) plays an important role in the instruction of the cell cycle. In a variety of transformed cell lines, tumour protein is expressed in high amounts, and it is believed to contribute to transformation and malignancy. This research aimed to detect the anti-p53 antibodies in sera of patients with various malignant tumours and to evaluate the sensitivity and specificity of an enzyme-linked immunosorbent assay (ELISA). A case-control study was conducted on samples from 49 patients with various types of malignant tumours at Sultanah Bahiyah Hospital, Alor Setar, Kedah, Malaysia, and 32 healthy control cases with non‐malignant disease collected from Universiti Sains Malaysia clinic, Penang, Malaysia. The antibodies against p53 protein in the serum samples were analysed using the commercial ELISA kit, Calbiochem® p53- ELISAPLUS. The results showed that the rate of anti-p53 antibodies in patients with various malignant tumours was 13 out of 49 (26.5 %), compared with only 2 out of 32 (6.25%) in healthy controls (p < 0.001). The sensitivity of this kit reached 28.6% and the specificity was 93.8%. In conclusion, these results suggest that the anti-p53 antibodies can be detected in different sera of malignant tumour patients and the ELISA kit is highly specific; nevertheless, its discrimination power is not perfect because of its low sensitivity to determine the anti-p53 antibodies.

Uselessness of Serum p53 Antibody for Detecting Colitis-associated Cancer in the Era of Immunosuppressive Therapy.

BACKGROUND/AIM: The present study examined the utility of serum p53 antibody (Ab) for detecting colitis-associated cancer (CAC) in the era of immunosuppressive therapy. PATIENTS AND METHODS: Two hundred and fifty patients were analyzed, 219 had no carcinoma or dysplasia (Group non-CAC), and 31 had carcinoma or dysplasia (Group CAC). Serum p53 Abs were detected with an enzyme-linked immunosorbent assay. Immunohistochemical detection was performed in Group CAC. RESULTS: Immunosuppressive therapy was performed in 98.1% of Group non-CAC and 80.6% of Group CAC. There were no differences in serum p53 Abs positivity between Groups non-CAC and CAC (8.7% vs. 3.2%, p=0.30). p53 staining positivity was noted in 90.3% of Group CAC, and the rate of serum p53 positivity was significantly lower in patients with immunosuppressive therapy than in those without in Group CAC (0.0% vs. 16.7%, p=0.04). CONCLUSION: The utility of serum p53 Ab for detecting CAC is dubious in the era of immunosuppressive therapy.

Sera Anti-P53 Antibody Provides New Information Which Explains the Link Between Diabetes and Cancer.

INTRODUCTION: Abnormalities in glucose metabolism in diabetic patients may lead to an increased risk of certain cancers. Epidemiological studies and meta-analysis have shown that factors such as gender, age, obesity, and insulin resistance are related to cancer incidence. The anti-p53 antibody is a known cancer marker due to tumor-associated p53 accumulation. Many studies have aimed to unravel the link between diabetes and cancer. Here, we aimed to elucidate the impact of diabetes on malignancies by analyzing anti-p53 antibody in sera of type 2 diabetes mellitus (T2DM) patients. MATERIALS AND METHODS: We conducted an observational study with a cross-sectional design. A total of 149 subjects comprised of 78 T2DM patients (32 with cancer risk and 46 subjects without cancer risk), 51 T2DM patients with cancer, and 20 healthy subjects as controls from multisites. The anti-p53 antibody was measured by enzyme-linked immunosorbent assay, while HbA1c was measured using the NGSP standardized method. RESULTS: We observed an 8.3-fold (p<0.05) increase of anti-p53 antibody in the sera of T2DM patients and a 24-fold increase (p<0.001) in T2DM patients with cancer compared to healthy subjects. The anti-p53 antibodies significantly increased almost three times (p<0.05) in T2DM patients with cancer (0.72 U/mL±0.20) compared to T2DM patients (0.25 U/mL±0.05). Meanwhile, this antibody was almost undetectable in healthy subjects as a control group (0.03 U/mL±0.03). The anti-p53 antibody level was higher in T2DM with cancer risk patients. However, we did not find a significant difference for it in T2DM without cancer risk patients (0.19 U/mL±0.03) and T2DM with cancer risk patients (0.29 U/mL±0.08). Multivariate regression analysis showed that T2DM with cancer was the only one independent factor (beta=0.218, p=0.019) that could predict the increase of anti-p53 antibody, controlled by age, gender, BMI, DM duration, and HbA1c. CONCLUSION: Our results showed that anti-p53 antibody almost not detected in healthy subjects, but 8.3-fold increase in the sera of T2DM patients and 24-fold increase in T2DM patients with cancer. Therefore, this biomarker provides new information which explains the link between diabetes and cancer.

Detection of p53 Mutations in Circulating DNA of Transplanted Hepatocellular Carcinoma Patients as a Biomarker of Tumor Recurrence.

p53 is the most commonly mutated gene in malignant human cancers. To detect p53 mutations in circulating DNA (cirDNA) of transplanted hepatocellular carcinoma (HCC) patients could be an interesting approach to know of any tumor recurrence. In this study, our objective was to determine the utility of this method in the diagnosis and the prognosis of HCC tumor recurrence.Twenty four liver transplanted HCC patients were included in the study together with a group of healthy controls. Detection of the specific p53 mutation in cirDNA was performed by high-resolution melting PCR (HRM-PCR) and COLD-PCR immediately before the transplantation. Serum anti-p53 was also determined using a p53-autoantibody ELISA kit.The results of the HRM-PCR and COLD-PCR showed two well-differentiated groups of transplanted patients after normalization by healthy controls. These data allow us to distinguish between patients with p53 mutated cirDNA and those with wild type cirDNA. Moreover, we have found that most of p53 mutated patients also presented elevated anti-p53 antibodies. The present results indicate that it is possible to detect mutated p53 genes with the cirDNA and that this could be used as a biomarker of tumor recurrence during the clinical evolution of the transplanted patients.

Clinical Implication of Serum Anti-p53 Antibodies and Overexpression of p53 Protein in Myelodysplastic Syndrome and Acute Myeloid Leukemia / 대한혈액학회지

BACKGROUND: Alteration of p53 genes is rare, but it is related with progressive diseases in hematologic malignancies. The wild type of p53 protein is not usually detected, but mutated p53 proteins are accumulated in the nuclei of tumor cells, which can be detected by immunohistochemical stain. Anti-p53 antibodies are found in the sera of patients with various malignant tumors as the result of immune response to accumulation of mutated p53 protein in tumor cells. METHODS: The relation of serum anti-p53 anti-bodies and cellular p53 protein expression to clinical features in 36 cases of myelodysplastic syndrome (MDS) and 58 cases of acute myeloid leukemia (AML) was analyzed. Anti-p53 antibodies in the patient's sera were measured with enzyme immunoassay (p53 autoantibodies ELISA, Dianova, Hamburg, Germany). Immunohistochemical staining for p53 protein was performed with the streptavidin-biotin-peroxidase method (LSAB kit, DAKO, Denmark) and anti-p53 monoclonal antibody (DO-7, DAKO, Denmark) in paraffin embedded bone marrow biopsy sections. RESULTS: Anti-p53 antibodies were positive in one of 36 (2.7%) MDS cases, and in four of 58 (6.8%) AML cases. Overexpression of p53 protein was seen in five (13.9%) of MDS and in five (8.6%) of AML. Serum p53 antibodies and p53 protein overexpression were more frequently found in RAEB, RAEB-t and M6 sutypes. There was no relation between anti-p53 antibodies and p53 protein overexpression in MDS and AML. The patients of MDS with anti-p53 antibodies and p53 overexpression tended to have severe dyserythropoiesis, higher Bounemouth scores and poor prognostic karyotypes and associated with shorter survival duration as compared to those without anti-p53 antibodies and p53 overexpression (4+/-1 vs 26+/-4 months, P=0.007). The patients of AML with anti-p53 antibodies and p53 protein overexpression tended to have poor prognostic karyotypes and resistance to chemotherapy. CONCLUSION: Anti-p53 antibodies are rarely observed in sera of patients with MDS and AML, probably reflecting the relatively low incidence of p53 mutation. But the findings suggest that the presence of p53 alteration could be useful to predict resistance to chemotherapy and short survival in particular sutypes of MDS and AML.

Clinical Significance of p53 Protein Overexpression and Serum Anti-p53 Antibodies in Patients with Acute and Chronic Leukemia / 대한임상병리학회지

BACKGROUND: Mutations of p53 gene, rarely found in leukemia, result in accumulation of mutated p53 protein in the nuclei of tumor cells, which can be detected by immunohistochemistry. Lately, anti-p53 antibodies were found in the sera of patients who had solid tumors as a result of immune response to accumulation of mutated p53 protein in tumor cells. METHODS: For investigation of the clinical implication of cellular p53 protein overexpression and serum p53 antibody, immunohistochemical staining for p53 protein of B-5 fixed paraffin embedded bone marrow biopsies and enzyme immunoassay for the presence of anti-p53 antibodies of sera were performed simultanously; in 58 cases of AML, 34 cases of ALL, 11 cases of acute leukemia at relapse, 13 cases of CML in chronic phase and 5 cases of CLL. RESULTS: Overexpression of p53 protein was found in 9.1%(11/121) of all leukemias, with 8.6% of AML with predominance of M6, 5.9% of ALL, 18.2% of acute leukemia at relapse and 40% of CLL, but not found in CML. Serum anti-p53 antibodies were found in 5.8%(7/121) of all leukemias, with 6.9% of AML and 5.9% of ALL, 9.1% of acute leukemia at relapse, but not found in chronic leukemias. In AML and ALL, age, sex, hemoglobin, leukocyte count, platelet count and blast % were not related with p53 protein expression. The AML patients with p53 protein overexpression have more unfavorable karyotypes(complex karyotype, -5, -7 and t(10;11)), with shorter overall survival as compared to those without p53 protein overexpression. The presence of serum anti-p53 antibodies was not related with clinical findings of leukemias. CONCLUSIONS: The indications are that p53 gene alterations will contribute to disease development and progression in some specific patients with leukemia, due to the rare frequency of overexpression of p53 protein and serum anti-p53 antibodies in leukemia. Analysis of the p53 protein and serum p53 antibodies could screen p53 gene mutation and predict prognosis for some leukemias.