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.

Mitochondrial unfolded protein response, mitophagy and other mitochondrial quality control mechanisms in heart disease and aged heart.

Mitochondria are involved in crucial homeostatic processes in the cell: the production of adenosine triphosphate and reactive oxygen species, and the release of pro-apoptotic molecules. Thus, cell survival depends on the maintenance of proper mitochondrial function by mitochondrial quality control. The most important mitochondrial quality control mechanisms are mitochondrial unfolded protein response, mitophagy, biogenesis, and fusion-fission dynamics. This review deals with mitochondrial quality control in heart diseases, especially myocardial infarction and heart failure. Some previous studies have demonstrated that the activation of mitochondrial quality control mechanisms may be beneficial for the heart, while others have shown that it may lead to heart damage. Our aim was to describe the mechanisms by which mitochondrial quality control contributes to heart protection or damage and to provide evidence that may resolve the seemingly contradictory results from the previous studies.

Telomerase activity in breast cancer patients: association with poor prognosis and more aggressive phenotype.

Telomerase expression is an important mechanism of tumor unlimited replicative potential. The aim of this study was to evaluate prognostic impact of telomerase activity in breast cancer patients and to correlate telomerase activity with established prognostic factors. We analyzed tissue of 102 malignant breast lesions and 20 healthy breast tissues. Telomerase activity was determined by telomeric repeat amplification protocol assay. Telomerase activity was present in 77 (75.49 %) of 102 breast cancers. Telomerase activity in breast cancers was statistically significantly higher in comparison with the activity in normal breast tissue. The levels of telomerase activity were significantly positively correlated with tumor size, axillary nodal status, histological grade, HER-2/neu protein expression in tumor tissue and expression of the nuclear antigen Ki-67. A statistically significant negative correlation was found between the presence of ER and telomerase activity. There was no correlation between telomerase activity and concentration of PR or the age of patients. Kaplan-Meier analysis showed that patients with higher telomerase activity had significantly shorter 10-year disease-free survival (p < 0.0001) and 10-year overall survival (p < 0.0001) than those with lower telomerase activity. These results were confirmed by logistic regression analysis. Our results support the prognostic role of telomerase activity and its relationship with the more aggressive phenotype of breast cancer.

Circulating Her-2/neu extracellular domain in breast cancer patients-correlation with prognosis and clinicopathological parameters including steroid receptor, Her-2/neu receptor coexpression.

HER-2/neu extracellular domain (ECD) can be detected in blood as a soluble circulating protein. The aim of this study was to analyze the relationship between HER-2/neu extracellular domain in the serum and the prognosis in breast cancer patients. We also correlated HER-2/neu ECD with various clinicopathological factors including steroid receptor, HER-2/neu receptor coexpression. The serum from seventy nine patients with invasive breast cancer and twenty individuals without malignancy was analyzed using the enzyme-linked immune adsorbent assay method. The cut-off value was estimated by the ROC curve analysis (15.86 µg/L). HER-2/neu ECD values in the serum of patients with breast cancer were significantly higher than in control subjects. Circulating HER-2/neu ECD was significantly associated with the histological grade of tumors and the status of axillary lymph nodes. Negative correlation was observed between HER-2/neu ECD in the serum and estrogen receptor positivity. When we analyzed HER-2/neu ECD in relation with coexpression of steroid receptor and HER-2/neu receptor in tissue, statistically higher values were found in the subgroup of patients with steroid receptor negative, HER-2/neu negative tumors than in the other subgroups. HER-2/neu ECD was not an independent factor in the univariate and multivariate analysis. However, elevated HER-2/neu ECD levels were found in patients with breast cancer possessing more aggressive phenotype.

Prognostic value of ki-67 in breast carcinoma: tissue microarray method versus whole section analysis- potentials and pitfalls.

In our study we have compared the prognostic value of two distinct methods of immunohistochemical Ki-67 determination, tissue microarray (TMA) and classical whole section analysis. "Cut-off" values were used according to the 2009 St. Gallen Consensus. Tissue specimens were obtained from a consecutive retrospective series of 215 female patients with primary invasive tumours. Two hundred and thirteen patients were included in the study. Data on Ki-67 was collected by both tissue microarray (TMA) and whole section analysis. Follow up data on overall (OS) and disease-free survival (DFS) were collected. Median follow-up was 95 months (range from 7.8 through 107 months). Mutual correlation of two Ki-67 determination methods was non-significant (Person's r = 0.13417; p = 0.0528). There was statistically significant association of whole section Ki-67 expression with histological and nuclear grade, progesterone receptor and HER2/neu status. The expression of Ki-67 protein in TMAs correlated only with histological and nuclear grade, but not with other traditional clinicopathological factors. Statistically significant differences in DFS (p = 0.0156) and OS (p = 0.0028) were confirmed between subgroups with low and high whole section Ki-67 expression. When subgroups with high and intermediate expression were compared, significant difference was found in DFS (p = 0.0272), but not in OS (p = 0.0624). On the other hand, there was no statistically significant difference either in DFS, or in OS, according to the expression of Ki-67 in TMAs (p = 0.6529; p = 0.7883; p = 0.7966 for DFS, and p = 0.8917; p = 0.6448; p = 0.4323 for OS, respectively). In our study, classical whole section was superior to TMA analysis in terms of prognosis and clinicopathological correlation. Our results indicate that the method used may have impact on prognostic significance of Ki-67. Further studies are needed, covering a greater number of patients and including a precisely defined stage and treatment patient cohorts, in order to solve controversies in Ki-67 assessment methodology.

Prognostic significance of urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor (PAI-1) in patients with primary invasive ductal breast carcinoma - a 7.5-year follow-up study.

AIMS AND BACKGROUND: Urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor (PAI-1) are key molecules in pericellular proteolysis, a process that plays an important role in tumor invasion and metastasis. In the current study we investigated the prognostic significance of uPA and PAI-1 in primary invasive breast cancer. METHODS AND STUDY DESIGN: uPA and PAI-1 antigen levels were determined by enzyme-linked immunosorbent assay in cytosols of 177 invasive ductal carcinoma specimens. The prognostic significance of uPA and PAI-1 was assessed for overall survival. The median follow-up time was 90 months. RESULTS: In univariate analysis, both uPA (third versus first tertile range of values; P = 0.02; HR = 2.08) and PAI-1 (third versus first tertile; P = 0.0007; HR = 3.1) were significant prognostic markers for overall survival. In multivariate analysis only nodal status (N2 vs N0; P = 0.0001; HR = 3.94) and PAI-1 (third versus first tertile; P = 0.004; HR = 3.05) remained significant independent prognostic factors. Both uPA and PAI-1 were correlated with established prognostic markers including histological grade, tumor size and Nottingham index. CONCLUSION: Our study with a 7.5-year follow-up confirmed the relation between elevated uPA and PAI-1 values and an aggressive course of invasive breast cancer. The prognostic significance of PAI-1 as an independent marker was proved for the overall group of breast cancer patients and the subgroup of node-positive patients.

TNF alpha promoter polymorphisms analysis in benign and malignant breast lesions.

Polymorphisms in genes involved in the complex mechanisms of carcinogenesis may affect the susceptibility to cancer. The multifunctional cytokine tumor necrosis factor alpha (TNF alpha) has an important role in the pathogenesis of inflammatory, autoimmune and malignant diseases. It has a large spectrum of activities, including both antitumorigenic and protumorigenic. In recent years, several TNF alpha promoter polymorphisms have been identified and related to the expression level of cytokine and to the susceptibility to solid tumors. The aim of our study was to investigate the frequency of three TNF alpha promoter polymorphisms (-1031, -308 and -238) in benign (fibrocystic changes) and malignant (invasive carcinoma) breast lesions. Using "real-time" PCR SNP analysis these polymorphisms were determined in 76 patients with benign and 158 patients with malignant breast lesions. The high expression genotypes at any of the three SNP polymorphisms were more frequent in invasive breast carcinoma (in 81 of 158 examined, 51.3%) than in fibrocystic changes (in 33 of 76 examined, 43.4%). The combined frequency of high production genotypes (-1031 T/C and C/C, -308 G/A and A/A and -238 G/A and A/A) was higher in patients with invasive breast carcinoma than in those with fibrocystic changes. However, these results were not statistically significant. Further studies on a larger group of patients are needed to evaluate the significance of potential differences in TNF alpha genotypes in different breast lesions.

[Pathophysiology of ischaemia-reperfusion injury]. / Patofiziologija ishemijsko-reperfuzijske ozljede.

Reperfusion of ischaemic tissue provides oxygen and substrates that are necessary for tissue recovery and concurrently removes toxic metabolites. However, reperfusion may induce various detrimental processes that may cause further tissue damage. Such deterioration of tissue function after reperfusion is defined as ischaemia-reperfusion injury. The consequences of ischemia-reperfusion injury vary from reversible cell dysfunction to local and remote tissue destruction, multiple organ failure and death. The pathogenesis of ischaemia-reperfusion injury is complex and includes excessive production of reactive oxygen species, activation of neutrophils, activation of complement, involvement of cytokines and other inflammatory mediators, vasoactive substances NO and endothelin. This review discusses the pathophysiology of ischaemia-reperfusion injury, the mechanisms of reactive oxygen species production, and the role of other factors in the pathogenesis of such injury. Several approaches and procedures used in pre-clinical and clinical studies in order to limit ischaemia-reperfusion injury are also presented.

[Gastrointestinal stromal tumors (GIST)--a paradigm of successful targeted therapy of solid tumors]. / Gastrointestinalni stromalni tumori (GIST)--paradigma uspjesnog ciljanog lijecenja solidnih tumora.

Gastrointestinal stromal tumors (GIST) have attracted basic scientists as well as clinicians in the last 10 years. The reason for this is explanation of the pathogenetic mechanism of tumor growth by activation of c-Kit protein, followed by a rationally designed suppressor, a drug named imatinib. It is the first successful therapy for solid tumors to date, although there are other ongoing studies of agents with targeted action on different molecules in different tumors. In 80% of patients there is a clinical benefit from imatinib trreatment. GIST shows great diversity in clinical presentation and some questions still remain, such as malignant potential and prognostic criteria in these tumors. Imatinib therapy demonstrates many favorable effects such as acceptable toxicity and relative mild side effects, excellent quality of life, good patient compliance, etc. There are ongoing trials of new agents designated for target molecules, which would hopefully show benefit after developing resistance to imatinib.

Expression of c-myc, erbB-2, p53 and nm23-H1 gene product in benign and malignant breast lesions: coexpression and correlation with clinicopathologic parameters.

The aims of this study were to assess the expression of protein products of c-myc, erbB-2, p53 and nm23-H1 gene in benign and malignant breast lesions, to estimate their possible coexpression and to correlate the results of immunohistochemical analysis with various clinicopathologic parameters. The method used was the immunohistochemical detection of the corresponding protein. Expression of c-myc protein was high in both malignant and benign lesions (95% and 100%). Expression of erbB-2 and mutated p53 proteins in malignant lesions was 27% and 34%. These proteins were present in benign lesions as well: 7.8% of benign lesions were positive for erbB-2 protein and 19.6% for p53 protein. The expression of nm23-H1 protein was similar in benign and malignant lesions: 47% and 54%. The coexpression of nm23-H1 and mutated p53 protein was found in 14 carcinomas (16.5%). We found a tendency of negative correlation between the expression of these two proteins. We also found a negative correlation between the size of breast carcinomas and the expression of nm23-H1, a higher proportion of nm23-H1-positive carcinomas in the group of erbB-2-negative, p53-negative carcinomas and a higher proportion of nm23-H1-positive carcinomas in the group of malignant lesions with negative axillary lymph nodes. Our results support the hypothesis that in women with breast cancer the expression of nm23-H1 gene may contribute to more favorable phenotype. We also showed that some changes found in malignant breast tumors such as the presence of mutated p53 protein and the expression of erbB-2 protein may be found in benign lesions as well.