Joanna Reszec

Expression of the Insulin‐Like Growth Factor‐I Receptor and Proapoptotic Bax and Bak Proteins in Human Colorectal Cancer

Abstract: Insulin‐like growth factor‐I (IGF‐I) and IGF‐I receptor (IGF‐IR), despite their well‐known roles in cell survival and proliferation, can also weakly enhance apoptosis. To study the relationships between the IGF‐IR and Bax as well as Bak, 144 cases of colorectal cancer were examined by immunohistochemistry, using the avidin‐biotin‐peroxidase method. The results were correlated with selected clinicopathological features of colorectal cancer and with the expression of IGF‐IR, Bax, and Bak in normal colon mucosa. In Bax‐, Bak‐, or IGF‐IR‐positive cancers, the adjacent colorectal mucosa revealed positive immunostaining for these proteins. In the majority of Bax‐, Bak‐, or IGF‐IR‐negative tumors, we observed no staining for these proteins in adjacent mucosa. The strong immunostaining for IGF‐IR, Bax, and Bak was noted in 50.8, 55.5, and 49.3% of tumors, respectively. We observed positive correlations between IGF‐IR and Bax (P < 0.002, r= 0.302), between IGF‐IR and Bak (P < 0.0001, r= 0.407), and between Bax and Bak (P < 0.0001, r= 0.474). No relationship was noted between IGF‐IR expression and tumor grade, stage, or lymph node status. We found negative associations between Bax, Bak, and tumor grade (P < 0.01 and P < 0.003, respectively), but no relationships between Bax and Bak and tumor stage or between Bax and Bak and lymph node status. Our results demonstrate that, in addition to overexpressed IGF‐IR, there are relationships between IGF‐IR and proapoptotic proteins in colorectal carcinomas that could contribute to increased cell turnover and the progression of colorectal cancer.

Expression of Cell Proliferation and Apoptosis Markers in Papillomas and Cancers of Conjunctiva and Eyelid

Abstract: Cell proliferation and programmed cell death are considered to be important events in carcinogenesis. The object of our study was to evaluate the expression of the Bcl‐2 protein family (Bcl‐2, Bak, Bax), p53, proliferating cell nuclear antigen (PCNA), and Ki‐67 protein immunoexpression as well as the correlation between the examined markers and some clinicopathological features in papillomas and cancers of conjunctiva and eyelid. Forty‐five squamous cell papillomas (SCP), 11 squamous cell cancers (SCC), and 27 basal cell cancers (BCC) were estimated. In the SCP group, p53 protein expression was observed in 30 cases (66.6%), Ki‐67 in 14 (31.1%), PCNA in 44 (97.8%), Bcl‐2 in 24 (53.3%), Bak in 28 (62.2%), Bax in 31 (68.9%), and Bcl‐xl in 11 (100%). In the SCC group, p53 protein expression was evaluated in 8 cases (72.8%), Ki‐67 in 2 (18.2%), PCNA in 8 (72.7%), Bcl‐2 in 5 (45.4%), Bax and Bak both in 10 (90.9%), and Bcl‐xl in 100%. In the BCC group, p53 protein expression was estimated in 23 cases (85.1%), Ki‐67 in 13 (48.1%), PCNA in 26 (96.2%), Bcl‐2 in 13 (48.1%), Bak in 21 (77.8%), Bax in 22 (81.5%), and Bcl‐xl in 23 (85.2%). We observed a correlation between some clinicopathological features and the examined markers of apoptosis and cell proliferation, which seemed to be important events in cancer development.

Expression of MMP-9 and VEGF in meningiomas and their correlation with peritumoral brain edema.

Meningiomas constitute up to 13% of all intracranial tumors. The predictive factors for meningioma have not been unambiguously defined; however some limited data suggest that the expression of matrix metalloproteinases (MMPs) and vascular endothelial growth factor (VEGF) may be associated with the presence of peritumoral brain edema (PTBE) and worse clinical outcome. The aim of this study was to analyze the expressions of MMP-9 and VEGF in a group of meningiomas of various grades and to study associations between these two markers and PTBE. The study included patients with supratentorial meningiomas. The patients were divided into low- (G1) and high-grade meningiomas (G2 and G3). PTBE was assessed on MRI. The expressions of VEGF and MMP-9 were determined immunohistochemically. The expression of MMP-9 was observed significantly more often in G3 meningiomas than in lower grade tumors. The presence of stage II or III PTBE was associated with a significant increase in MMP-9 expression. The expression of VEGF did not differ across the PTBE stages. Our findings point to a significant role of MMP-9 and VEGF in the pathogenesis of peritumoral brain edema in low- and high-grade meningiomas.

Evaluation of Apoptosis Markers in Conjunctival and Eyelid Benign and Malignant Tumors

Abstract: The balance between cell proliferation and programmed cell death plays a crucial role in malignant development. Bcl‐2 family proteins, including proapoptosis protein Bak and antiapoptosis protein Bcl‐2, regulate the apoptotic process. Mutation of the p53 gene, which results in P53 protein accumulation, was observed in many types of human cancer. The aim of our study was to evaluate immunohistochemical Bcl‐2, Bak, and P53 protein expression and the relation between these proteins in conjunctival and eyelid benign and malignant tumors. We examined a series of 42 papillomas (CEP), 12 squamous cell cancers (SCC), and 19 cases of basal cell cancer (BCC). The age in the CEP group ranged from 18–94 years, and in the SCC and BCC groups from 42–87 years. Staining patterns were correlated with sex, age, and tumor localization. P53 protein‐positive immunostaining was observed in 71% of cases, Bcl‐2 in 83.9%, and Bak in 74.2 cases in the SCC and BCC groups. In the CEP group, P53 overexpression was observed in 90.5% of cases, Bcl‐2 in 71.4%, and Bak in 76.2%. No statistically significant correlation was found between examined protein expression and sex, age, and tumor localization. An inverse correlation was observed between P53 and Bak protein expression in the CEP group. No statistically significance correlation was noted between Bcl‐2 and P53 and Bcl‐2 and Bak protein expression in both examined groups. The obtained data suggests that P53 and Bcl‐2 protein expression coupled with decreasing Bak expression are associated with apoptosis and proliferation as well as malignant progression in conjunctival and eyelid tumors.

Validation for histology-driven diagnosis in non-small cell lung cancer using hsa-miR-205 and hsa-miR-21 expression by two different normalization strategies.

Targeted therapy of non‐small cell lung cancer (NSCLC) demands a more accurate tumor classification that is crucial for patient selection in personalized treatment. MicroRNAs constitute a promising class of biomarkers and a helpful tool for the distinction between lung adenocarcinoma (AC) and squamous cell lung carcinoma (SCC). The aim of this study was to evaluate the impact of two different normalization strategies, using U6 snRNA and hsa‐miR‐103 as reference genes, on hsa‐miR‐205 and hsa‐miR‐21 expression levels, in terms of the classification of subtypes of NSCLC. By means of a quantitative real‐time polymerase chain reaction (qRT‐PCR) microRNA expression levels were evaluated in a classification set of 98 surgically resected NSCLC fresh‐frozen samples, and validated findings in an independent set of 42 NSCLC samples. The microRNA expression levels were exploited to develop a diagnostic test using two data normalization strategies. The performance of microRNA profiling in different normalization methods was compared. We revealed the microRNA‐based qRT‐PCR tests to be appropriate measures for distinguishing between AC and SCC (the concordance of histologic diagnoses and molecular methods greater than 88%). Performance evaluation of microRNA tests, based on the two normalization strategies, showed that the procedure using hsa‐miR‐103 as reference target has a slight advantage (sensitivity 83.33 and 100% in classification and validation set, respectively) compared to U6 snRNA. Molecular tests based on microRNA expression allow a reliable classification of subtypes for NSCLC and can constitute a useful diagnostic strategy in patient selection for targeted therapy.

Assessment of inflammatory infiltration and angiogenesis in the thrombus and the wall of abdominal aortic aneurysms on the basis of histological parameters and computed tomography angiography study

The proliferation of vessels within the aneurysms wall and the intraluminal thrombus of abdominal aortic aneurysm (AAA) may be the main factor responsible for progression and rupture of AAA. The aim of this study was to compare the parameters of the thrombus (size, density, contrast enhancement) measured by com- puted tomography (CT) with histological assessment of thrombi removed during surgery. 29 patients with AAA were examined with angio-CT. Post-surgery histopathological evaluation of AAA was performed. Slides were stained with markers of B- (CD20) and T-lymphocytes (CD3), and markers of endothelial cells (CD34). Throm- bi were enhanced after contrast media administration in angio-CT (p = 0.002). There was a statistically signifi- cant correlation between contrast enhancement and the presence of B lymphocytes. Intensity of endothelial cell marker expression significantly correlated with the presence of inflammatory T- and B-cells. No statistical signif- icant correlation was found between contrast enhancement of the thrombus and markers of endothelial cells. The accumulation of inflammatory cells in the wall of AAA thrombus results in the formation of new vessels which participates to the instability of the thrombus and AAA wall. Assessment of the inflammation and neovas- cularization in the wall and thrombus of the AAA might be an important factor in monitoring the progression and the risk of aneurysms rupture. (Folia Histochemica et Cytobiologica 2012, Vol. 50, No. 4, 547-553)

The expression of hypoxia-inducible factor-1 in primary brain tumors

Purpose: Primary brain tumors are common type of neoplasms. The most common are astrocytic tumors, so do meningiomas of various grades. The etiology is still unknown; however, there are lots of data presenting new theories about genetic alterations responsible for low- or high-grade astrocytic tumors development as well as meningiomas, despite this the results are divergent. The aim of the study was to evaluate hypoxia-inducible factor-1 (HIF-1) expression in meningiomas and astrocytic tumors of various grades. Material and methods: One hundred six cases of astrocytic tumors were divided into diffused astrocytoma (24 cases), anaplastic astrocytoma (40 cases) and glioblastoma groups (42 cases). Among glioblastoma group, 30 cases were secondary glioblastoma. One hundred fifty-four meningioma cases were divided as low-grade meningioma (G1: 104 cases) and high-grade meningioma groups (G2: 43 cases and G3: 7 cases). Twelve low-grade meningiomas transformed into high-grade tumors, 17 low-grade meningiomas recur within 12 years. HIF-1 expression was estimated using immunohistochemistry under the light microscope. Statistical analysis was performed in all examined groups. Results: HIF-1 expression was observed in 37.5% cases of diffused astrocytomas, in anaplastic astrocytomas 27.5% tumors were HIF-1 positive, in the glioblastoma goup HIF-1 expression was observed in 83.3% cases. All secondary glioblastomas were positive for HIF-1. Low-grade meningiomas were positive for HIF-1 in 55.7%, in high-grade meningiomas, HIF-1 expression was observed in 84%. All meningiomas, which progressed from low- to high-grade meningiomas, were HIF-1 positive. Conclusion: HIF-1 expression is associated with the development and progression of both astrocytic tumors and meningiomas.

Is littoral cell angioma of the spleen as rare as previously believed in the pediatric population

Littoral cell angioma (LCA) is a rare primary splenic vascular tumor, originating from the littoral cells lining the red pulp sinuses of the spleen. There are only a handful of case reports of LCA in children to be found in the literature. We performed a retrospective analysis of the medical charts of pediatric patients with splenic lesions who were treated between 2005 and 2010 in the Pediatric Surgery Department of the Medical University of Bialystok. Surprisingly, LCA accounted for 37.5% of the splenic lesions found in our series. The majority of LCA tumors are benign, but given their malignant potential, splenectomy and long-term follow-up should be the gold standard for their management. We strongly support the use of further cross-sectional studies to properly elucidate the prevalence of littoral cell angioma of the spleen in the pediatric population.

Survivin, caspase-3 and MIB-1 expression in astrocytic tumors of various grades

PURPOSE Gliomas are the most common primary brain tumors. The etiology is still unclear and the progression from low to high-grade gliomas is frequent. The molecular mechanisms are quite established, however the heterogeneity of glioblastomas force the scientist to look for the new therapeutic targets. The aim of the study was to evaluate the caspase-3 and survivin expression in correlation with MIB-1 expression in gliomas of various grade to assess the apoptosis in gliomas and to determinate new possible targets for the future therapy. MATERIAL AND METHODS We identified 131 patients with a histopathological diagnosis of astrocytic tumors (diffuse astrocytoma, anaplastic astrocytoma and glioblastoma). The evaluation of caspase-3, survivin and MIB-1 expression was done using immunohistochemical methods. RESULTS Caspase-3 and survivin expression was observed both in low- and high-grade astrocytomas. The differences in expression were the most evident in glioblastoma group. All primary glioblastomas (31 cases) expressed caspase-3. In secondary glioblastoma group only 17 out of 30 specimens were positive for caspase-3. Survivin expression was observed in 80.6% primary glioblastomas and in all examined secondary glioblastomas and the staining was strong and diffuse in all cases. MIB-1 expression was low in diffuse astrocytomas (DA) and ranged between 1 and 5%. In anaplastic astrocytoma group it was ranged between 5 and 10% and the highest percentage of the positive cells was observed in glioblastoma cases and ranged from 10% even to 30%. The most evident MIB-1 expression was observed in the cells surrounding the pathological blood vessels and necrosis. CONCLUSIONS The high incidence of survivin and caspase-3 expression in diffuse and anaplastic astrocytoma cases may suggest, that the regulation between pro- and antiapoptotic proteins may play an important role in tumor growth and progression. The overexpression of survivin and MIB-1 expression in glioblastoma cases also may confirm the theory about the important role of anti-apoptotic and proliferation processes in glioblastoma progression and as such may be potential therapeutic targets.

c-erbB-2 protein expression in astrocytic tumors of the brain

Summary Background Astrocytic tumors are the primary brain tumors, which often progress to glioblastoma, a highly malignant neoplasm of the central nervous system. There is much new data regarding to the formation and progression of these tumors; however, glioblastoma remains one of the most fatal neoplasms in humans. The aim of the study was to evaluate the role of c-erbB-2 protein expression in various groups of astrocytic tumors. Material/Methods 65 cases of astrocytic tumors were divided into 3 groups: diffuse astrocytoma (group I; n=17 cases), anaplastic astrocytoma (group II; n=23 cases) and glioblastoma (group III; n=25 cases). C-erbB-2 protein expression was estimated semiquantitatively on immunohistochemically stained tissue sections using antibodies against c-erbB-2 protein. Statistical analysis was performed in all examined groups. Results The c-erbB-2 protein expression was observed in 15 out of 17 cases (88.3%) in group I, 22 out of 25 cases (88%) cases in group II, and in 19 out of 23 cases (82.6%) in group III. There were no statistically significant differences between the examined groups. The strongest c-erbB-2 immunoexpression was observed in low grade astrocytomas (diffuse astrocytomas G2); in the glioblastoma group the c-erbB-2 protein expression was weak and 17.4% of cases were negative. Conclusions C-erbB-2 protooncogene alteration is an early phenomenon in glial tumor development and progression.