Expression of insulin-like growth factor receptor type 1 correlate with lymphatic metastases in human gastric cancer.

Most patients with gastric cancer are diagnosed at advanced clinical stages with a high frequency of lymph node metastasis. It is very important to find novel factors for the early diagnostic and prognostic evaluation of gastric cancer. It has been shown that IGF-1R activates mitotic division and inhibits apoptosis of cancer cells through the activation of signaling MAP/ERK and PI3K/Akt-1 pathways. IGF-1R plays a role in cell transformation and maintenance of the phenotype in modified cells. Moreover, an IGF-1 receptor effect influences the processes of adhesion, migration, invasion and metastasis of tumor cells. The aim of the study was to assess the expression of IGF-1R in gastric carcinoma in correlation with selected anatomo-clinical parameters. The study enrolled a group of 49 patients treated surgically for gastric cancer. 28 patients had no lymph node metastases. The expression of the studied proteins was assessed using the immunohistochemical method. We found that the expression of IGF-1R in gastric cancer is associated with lymph node metastasis (p < 0.001), is correlated with worse prognosis and high histological malignancy grade, and is an independent predictor of survival in patients with gastric cancer (p < 0.001). IGF-1R may play an important role in tumor growth and metastasis via the lymphatic pathway.

Diagnostic significance of TIMP-1 level in serum and its immunohistochemical expression in colorectal cancer patients.

Tissue inhibitor of metalloproteinase-1 (TIMP-1) inhibits the ability of cancer cells to metastasize, but it can also stimulate cancer development. The aim of this study was to assess the level of TIMP-1 in serum and its expression in patients with colorectal cancer (CRC). The study group consisted of 43 patients diagnosed with colorectal cancer and 24 healthy volunteers. The level of TIMP-1 was assessed by the ELISA method while the expression of this protein was performed immunohistochemically. The concentration of TIMP-1 in the sera of colorectal cancer patients was significantly higher than in the healthy control group (p = 0.004). Higher level of TIMP-1 in the sera correlated with female gender (p = 0.045), tumor location in colon (p = 0.016), poorly differentiated tumor (p = 0.034) and higher platelet count in whole blood (p < 0.004). A positive reaction of the protein in cancer cells was observed in 31 cases and was found to correlate negatively with its reaction in peritumoral stroma (p < 0.001). According to this study, TIMP-1 protein may play an important role in cancer development. The assessment of this molecule in serum and tissue can be useful at the time of diagnosis and can help us to understand the nature of colorectal pathogenesis.

Expression of phosphatase of regenerating liver-3 (PRL-3) in endometrioid cancer and lymph nodes metastases.

PURPOSE: We identify the expression of PRL-3 in primary endometrioid endometrial cancer and metastases in relation to the clinicopathological characteristics. MATERIAL/METHODS: The study involved 30 patients with type I endometrial cancer. Twelve of them were diagnosed with metastases in various localization of abdomen. The PRL-3 expression was evaluated on the basis of immunohistochemistry results by the use of monoclonal antibody anti-PRL3 clone 3B6. RESULTS: The intensity of PRL-3 expression in correlation with tumor stage was statistically significant (p = 0.024). The strongest reaction was noted in cases classified as a 1a and 1b stage defined by FIGO. The strength of PRL-3 expression is significantly associated with the degree of histological tumor grade (p = 0.035). CONCLUSIONS: The strong expression of PRL-3 in the primary tumor that was significantly correlated with the grade and clinical stage suggest that PTP4A3 participates in the process of endometrial carcinogenesis.

Matrix metalloproteinase 2 (MMP-2) and their tissue inhibitor 2 (TIMP-2) in gastric cancer patients.

BACKGROUND: Matrix metalloproteinase 2 (MMP-2) is able to degrade type IV collagen and its activity is mostly regulated by tissue inhibitor of matrix metalloproteinase 2 (TIMP-2). These proteins might play a role in tumor progression, including gastric cancer (GC). METHODS: The study included 108 individuals, GC patients and healthy subjects. Serum levels of all analyzed markers were evaluated by the immunological methods, while immunohistochemistry was used to assess the expression of these proteins in GC, interstitial inflammatory cells and normal tissues. RESULTS: The percentage of positive reactions of MMP-2 and TIMP-2 was higher in GC and inflammatory cells compared to normal tissue, while serum levels of these proteins were statistically lower in GC patients in comparison to healthy subjects. There was a significant positive correlation between TIMP-2 immunoreactivity in inflammatory cells and the presence of lymph node metastasis. Area under ROC curve (AUC) for TIMP-2 was higher than MMP-2, while serum MMP-2 was an independent prognostic factor of GC patients' survival. CONCLUSION: Our findings suggest that TIMP-2 seems to be a predictor of tumor progression, especially for nodal involvement, whereas serum MMP-2 might be useful as an independent prognostic factor of patients' survival.

Immunohistochemical assessment of apoptosis-associated proteins: p53, Bcl-xL, Bax and Bak in gastric cancer cells in correlation with clinical and pathomorphological factors.

PURPOSE: The p53 protein as well as Bcl-2 family proteins such as Bax, Bak and Bcl-xL regulate apoptosis. The study objective was to analyze the expression of p53, Bak, Bcl-xL and Bax in gastric cancer and in healthy gastric mucosa. MATERIAL AND METHODS: The study group consisted of 66 patients with gastric cancer, treated surgically in II Department of General and Gastroenterological Surgery, Medical University of Bialystok. The expression of the studied proteins was assessed using the immunohistochemical method. RESULTS: Significant differences were found in the expressions of the studied proteins as compared to healthy gastric mucosa. The expressions of p53 and Bax were significantly higher (70% vs 13% and 50% vs 13%), whereas those of Bak and Bcl-xL significantly lower (18% vs 83% and 74% vs 97%) in cancer cells than in normal mucosa (p<0.001). Significant differences were also noted in the expressions of Bax and Bcl-xL in relation to histological type. In the intestinal type (Lauren I), the expressions of Bax and Bcl-xL were higher as compared to the diffuse type (Lauren II) (93% vs 43% and 91% vs 43%). Simultaneously, correlations were noted between changes in the expression of Bax vs Bcl-xL and Bak. High expression of Bax showed a positive correlation with reduced Bak and Bcl-xL (p<0.05). Moreover, positive expression of p53 caused poorer distant survival of patients (p<0.05). CONCLUSION: Our study concluded that disturbances in the expression of p53, Bax, Bcl-xL and Bak proteins are associated with their involvement in the process of carcinogenesis in the stomach. It is suggesting that they might appeared in the early phase of carcinogenesis.

Immunohistochemical assessment of PRL-3 (PTP4A3) expression in tumor buds, invasion front, central region of tumor and metastases of colorectal cancer.

PURPOSE: The aim of the study was to evaluate immunohistochemical expression of PRL-3 protein tumor buds, invasion front, central region of tumor and metastases of colorectal cancer. MATERIAL/METHODS: The PRL-3 expression was analyzed in 103 colorectal carcinoma patients, using the immunohistochemical method with a monoclonal antibody 3B6 anti-PRL-3 (Attogen Biomedical Research, USA). RESULTS: Positive reaction for PRL-3 was observed in 36.9% of cases in the central region of tumor, in 64.3% in the invasion front, and in as many as 81.4% in buds (present in 70/103 cases), in 100% in metastases to local lymph nodes, in 100% in metastases to the liver and in 97.1% in metastases to the lungs. The findings indicate that cancer cells obtain this protein already in the early stages of metastasizing. PRL-3 is present not only in metastases to local lymph nodes but also to distant organs. It is likely that PRL-3 protein takes part in the initiation of metastasizing of cancer cells. Also the presence of lymph and blood vessel invasion was found only to correlate with increased percentage of patients with strong PRL-3 expression in tumor buds (p=0.046). CONCLUSION: Our results may suggests the participation of PRL-3 protein as a marker of the presence of colorectal cancer metastasis to the lymph nodes and distant metastases, which is independent of parameters such as gender and age of the patients, tumor location, histological type and grade of histological malignancy and stage of tumor.

The EGFR expression in gastric mucosa of children infected with Helicobacter pylori.

PURPOSE: Epidermal growth factor receptor (EGFR) modulates balance between proliferation and apoptosis in gastric mucosa of the gastrointestinal tract. The aim of the study was to evaluate immunohistochemically the EGFR expression in epithelial and gland cells of antral mucosa in children infected with Helicobacter pylori (H. pylori). MATERIAL/METHODS: The study included 44 children, aged from 5 to 18 years (mean age 13+/-3.4 years) with dyspeptic symptoms, of whom 30 (68.2%) children were infected with H. pylori, 14 (31.8%) children constituted controls. Endoscopic and histopathological assessment of antral mucosa samples was performed according to the Sydney System. Samples taken from gastroscopy were prepared to evaluate EGFR expression in epithelial and gland cells of antrum mucosa according to the manual of a detection kit of EnVision+System-HRP (DAKO). RESULTS: In children H. pylori infected, the EGFR expression in epithelial cells of antral mucosa equaled on average 82.5+/-15 cells/mm2 and ranged from 45.0 to 98.0 cells/mm2 as well as differed statistically significantly when compared to controls (10.2+/-5.0 cells/mm2) (p<0.001). In children with H. pylori infection, the EGFR expression in gland cells of antral mucosa ranged from 2.0 to 85.0 cells/mm2 (mean 25.7+/-22.6 cells/mm2); was lower and differed statistically significantly from controls (54.2 +/- 29.6 cells/mm2) (p<0.001). In children H. pylori infected, there was a statistically significant difference (p<0.001) between the EGFR expression in epithelial and in gland cells of antral mucosa. CONCLUSION: The increased EGFR expression in epithelial cells in comparison with gland cells of antral mucosa in children with H. pylori infection may suggest its role in regeneration processes of gastric mucosa.

Functional categories of TP53 mutation in colorectal cancer: results of an International Collaborative Study.

BACKGROUND: Loss of TP53 function through gene mutation is a critical event in the development and progression of many tumour types including colorectal cancer (CRC). In vitro studies have found considerable heterogeneity amongst different TP53 mutants in terms of their transactivating abilities. The aim of this work was to evaluate whether TP53 mutations classified as functionally inactive (< or=20% of wildtype transactivation ability) had different prognostic and predictive values in CRC compared with mutations that retained significant activity. MATERIALS AND METHODS: TP53 mutations within a large, international database of CRC (n = 3583) were classified according to functional status for transactivation. RESULTS: Inactive TP53 mutations were found in 29% of all CRCs and were more frequent in rectal (32%) than proximal colon (22%) tumours (P < 0.001). Higher frequencies of inactive TP53 mutations were also seen in advanced stage tumours (P = 0.0003) and in tumours with the poor prognostic features of vascular (P = 0.006) and lymphatic invasion (P = 0.002). Inactive TP53 mutations were associated with significantly worse outcome only in patients with Dukes' stage D tumours (RR = 1.71, 95%CI 1.25-2.33, P < 0.001). Patients with Dukes' C stage tumours appeared to gain a survival benefit from 5-fluorouracil-based chemotherapy regardless of TP53 functional status for transactivation ability. CONCLUSIONS: Mutations that inactivate the transactivational ability of TP53 are more frequent in advanced CRC and are associated with worse prognosis in this stage of disease.

Evaluation of proliferating markers Ki-67, PCNA in gastric cancers.

Tumours from 45 patients with advanced gastric cancer were assessed by immunohistochemistry. Tissue sections were fixed in 10% buffered formaldehyde solution, embedded in paraffin and stained immunohistochemically with anti-human Ki-67 and PCNA antibodies. No correlation was found between Ki-67, PCNA protein expression, the age of patients and the localization of tumour. A significant, positive association was observed between the expression of Ki-67, PCNA and tumour differentiation and Lauren's classification. Also a strong correlation was found between lymph node involvement and the expression of Ki-67 protein. These data suggest that the expression of Ki-67, PCNA proteins were closely connected with the high grade of tumour malignancy.

Evaluation of protein products of cell cycle regulating genes in gastric cancer.

Formalin fixed, paraffin embedded tissue samples of 45 gastric carcinomas, resected curatively, were used for the study. An immunohistochemical analysis employed monoclonal antibodies: p53 (No N1581, DAKO) and p27KIP1 (NCL-p27KIP1, Novocastra). Positive nuclear protein expression was assessed at the 30% level. We found no correlations between the expression of either protein and Lauren's classification, the age of patients and tumour localization. Borderline significance of p=0.07 was noted in the association of p53 expression and histological differentiation. However, a decrease of p27 expression and an overexpression of p53 correlated with the presence of lymph node metastases (p<0.01). Simultaneously, the expression of p27 protein in main mass of tumour correlated with the lack of p53 expression in the main mass and lymph node metastases.

Effects of changes at the site of E-cadherin expression as an indicator of colon cancer aggressiveness.

The aim of the work was to study the effects of changes in the location of E-cadherin from membrane to cytoplasm and the appearance of metastases and recurrence in patients with colon cancer of pT1 grade. The study group consisted of 34 patients with colon cancer. The material was fixed in 10% buffered, directly following surgery, fixed in fonnaldehyde and embedded in paraffin blocks by a standard method. Immunohistochemical reactions were performed, using monoclonal E-cadherin antibodies (Novocastra, NCL-E-Cad). Statistical analysis did not show any relation between the change in the location of E-cadherin expression, the patients' sex, and the location of changes. Simultaneously, we observed a strong relationship between the presence of exudate in the vessels from cancer cells, the histological grade and the loss of E-cadherin expression in the main tumour mass (p<0.01). We also noted a statistically significant correlation between the presence of lymph node invasion and distant metastases and the E-cadherin cytoplasmic reaction (p=0.0001, p=0.000001, respectively). A borderline significance of p=0.06 was noted in the association between the appearance of recurrence at the postoperative site and the change in location of E-cadherin expression in the main tumour mass from cytoplasm to membrane. On the basis of our results, we can conclude that a change in the location of E-cadherin expression (from membrane cytoplasm) is strongly associated with an increased aggressiveness of CRC, which is related to the appearance of proximal and distant metastases and to recurrence at the postoperative scar.

Molecule CD44 variant 10 expression in lymphocytes infiltrating tumour tissues and epithelial cells in patients with colorectal cancer.

The aim of this study was to evaluate the expression of CD44v10 in colorectal tumour cells and in lymphocytes infiltrating the tumour (CD45+). Samples of tumour tissue (TT), as well as of healthy tissue (HT) and of tumour adjacent tissue (TAT), were obtained from 20 patients. An evaluation of CD44v10 expression was performed in a flow cytometer. The mean value of the percentage of CD45+ with co-expression of CD44v10 was significantly higher in the lower stage of the tumour (pT). The mean value of the percentage of epithelial cells with CD44v10 co-expression was significantly higher in pN2 than in pN1 stage. Only in TAT the mean value of the percentage of epithelial cells and CD45+ with tCD44v10 co-expression was significantly lower in the higher degree of histological malignancy. It is supposed that CD44v10 takes part in local cancer progression.

Proliferative activity of chosen central nervous system (CNS) neoplasms.

Gliomas are the most common neoplastic tumours of the central nervous system. The aim of the study was to evaluate the proliferative activity of chosen types of gliomas and to analyse their correlation with histological type, malignancy grade, location, size and clinical symptoms. The study involved patients with astrocytoma, anaplastic astrocytoma, glioblastoma, oligodendroglioma, anaplastic oligodendroglioma. The proliferative activity (the labelling index--LI) of glial cells was estimated, using immunohistochemistry. In studied groups, a positive correlation was noted between the proliferative activity and tumour size, but not between the proliferative activity and tumour location. The clinical symptoms were conditioned mainly by tumour location and, to a smaller extent, by its size.

P53 protein expression in oral squamous cell cancer in relation to some of its clinicopathological variables.

Oral squamous cell cancer develops through a multistep process by the accumulation of genetic and phenotypic changes. Loss of P53 tumor suppressor gene function represents the most common genetic lesion in human cancer. The significance of P53 expression for the development and progression of oral squamous cell cancer has still to be evaluated. The aim of this study was to estimate relationships between P53 protein expression and some clinicopathological variables of established or presumed prognostic value. A series of 129 oral squamous cell cancers was investgated retrospectively for expression of P53 protein by immunohistochemistry of paraffin-embedded tissue sections. The slides were stained with H+E and by immunohistochemistry with anti-human P53 antibody. Positive immunohistochemical staining for P53 protein was present in 75 (58%) oral cancer cases. There were no statistically significant correlations between oral cancer P53 expression and tumor site, grading, mitotic index, invasive margin type, as well as patients age and sex. Our results suggest that immunohistochemical overexpression of P53 is an important markerof accomplished neoplastic transformation in oral cavity lesions but it does not play a crucial role in the tumor progression.

Study of P53 protein expression in colorectal cancer.

Mutations of the P53 gene, strictly associated with the carcinogenesis are a commonly observed in neoplastic cells. The aim of this study was the immunohistochemical evaluation of P53 protein expression in colorectal carcinomas and analysis of its relationship to chosen anatomo-clinical and morphological parameters of the tumours. The study used the material obtained during surgical treatment of 74 colorectal carcinomas. Tissue sections were fixed in 10% buffered formaldehyde solution, embedded in paraffin and stained immunohistochemically with the antihuman P53 protein monoclonal antibody. The immunolocalization of P53 protein was performed using the Labelled Streptavidin Biotin (LSAB) method. The P53 protein expression was semiquantitatively assessed in neoplastic cells and the reaction present in more than 25% of tumour cells was accepted as the threshold of positivity. No correlation was found between P53 protein expression and tumour histologic type and site, and age and sex of patients. However, P53 protein expression in primary and metastatic tumours was found statistically significantly correlated.

Tumour budding intensity in relation to cathepsin D expression and some clinicopathological features of colorectal cancer.

Although it has been suggested that tumour budding at the invasive edge of colorectal cancer is an important prognostic factor its biological significance for tumour progression is still to be evaluated. The aim of the study was to correlate tumour budding intensity with cathepsin D expression and some other clinicopathological variables of presumed or established prognostic value. 48 patients with colorectal cancer at pT3 stage, G2 grade of histological differentiation and tumour budding at the invasive edge were evaluated. Colorectal tumours were investigated for cathepsin D expression by immunohistochemistry of formalin-fixed and paraffin embedded tissues. There was no statistically significant relationships between tumour budding intensity grade and primary tumour cathepsin D expression, stromal cell cathepsin D expression and histochemical immunostaining of cathepsin D in rumour budding at its invasive edge. The tumour budding intensity was not associated with lymph node status, tumour site, peritumoral inflammatory response as well as the patient's age and sex. The results of this study suggest that intensity of tumour budds formation at the invasive margin of colorectal cancer is not associated with presumed or established prognostic factors such as lymph node metastases, and peritumoural inflammatory reaction as well as cathepsin D expression.

Study of p53 protein expression in prostate cancer.

The aim of this study was the immunohistochemical evaluation of p53 protein expression in localized prostate cancer (Pca) following radical prostatectomy and analysis of its relationship to chosen anatomo-clinical and morphological parameters of the tumours. The present investigation included material from 28 randomly selected patients undergoing radical prostatectomy. Tissue sections were fixed in 10% buffered formaldehyde solution, embedded in paraffin and stained immunohistochemically with the anti-human p53 protein monoclonal antibody. The immunolocalization of p53 protein was performed using the Labelled Streptavidyn Biotin (LSAB) method. The p53 protein expression was semiquantitatively assessed in neoplastic cells and the reaction present in more then 25% of tumour cells was accepted as the threshold of positivity. No correlation was found between p53 protein expression and Gleason score, pT stage, lymph node metastases, seminal vesicles invasion, positive or negative surgical resection margins, age of patients. However, p53 protein expression and capsular penetration was found statistically significantly correlated.