Neutrophil infiltration combined with necrosis in the primary tumor is a useful prognostic indicator for three-year disease-free survival time in patients with colorectal cancer.

Histopathological evaluation plays a key role in the diagnosis of colorectal cancer (CRC). Tumor-related local inflammation is regarded as a novel prognostic parameter. Neutrophils constitute one of the main types of inflammatory cells. The aim of the present study was to evaluate the prognostic value of intratumoral tumor-associated neutrophils (intraTANs), stromal TANs (stromaTANs) and necrosis, as well as their combined parametric value in formalin-fixed paraffin-embedded tissue sections from patients with CRC. For this purpose, a retrospective study of 160 patients with CRC who underwent surgery was conducted. The association of intraTANs, stromaTANs, necrosis and their combined parametric value with the clinicopathological features of patients with CRC was examined. The Kaplan-Meier method and the log-rank test were used to compare survival curves. To identify independent prognostic factors, uni- and multivariate Cox proportional hazards regression models were used. StromaTANs were associated with lymph node metastasis (P=0.049) and tumor deposits (P=0.041). In addition, necrosis was found to be associated with venous (P=0.003), lymphatic (P=0.007) and perineural (P=0.015) invasion, as well as with lymph node metastasis (P=0.033), the number of invaded lymph nodes (P=0.012), and lymph node pouch invasion (P=0.043). Furthermore, necrosis was found to be associated with the white blood cell count (P=0.030), neutrophil count (P=0.011), the combined neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (NLR-PLR) (P=0.038), and the combined platelet and NLR (PLT-NLR status) (P=0.030), as well as with the serum carcinoembryonic antigen (CEA) levels following surgery (P=0.011) and the monocyte-to-lymphocyte ratio (P=0.023). The combined parametric value was found to be associated with pT stage (P=0.049), venous (P=0.034) and lymphatic (P=0.026) invasion, and with serum CEA levels prior to surgery (P=0.029). The analysis of the 3-year disease-free survival (DFS) time revealed that tumor growth [hazard ratio (HR), 2.070; 95% CI, 1.837-3.808; P=0.003] and the combined parametric value (intraTANs, stromaTANs and necrosis, HR, 1.577; 95% CI, 1.372-3.032; P=0.028) were independent factors for patients with CRC. Taken together, the findings of the present study demonstrated that the combined value of neutrophils and necrosis examined in the cancerous tissue may be used as a prognostic factor for the 3-year DFS time in patients with CRC.

STAT3 and apoptosis regulators: Bak and Bcl-xL in endometrioid adenocarcinomas of different estrogen receptor-α immunoprofile.

Estrogen receptor (ER) is a major feature of endometrioid adenocarcinoma. It has a significant impact on constitution of estrogen-responsiveness of this endometrial malignancy, in which STAT3 (signal transducer and activator of transcription) becomes hyperactivated. The aim of our study was to detect immunohistochemically and compare expressions of STAT3 with apoptosis regulators (Bak and Bcl-xL) in regard to different pathological features and variably pronounced ER-α immunoprofile in 78 endometrioid adenocarcinomas. STAT3 was abundantly detected in nuclei of cancer cells in 54 cases, thus pointing at its activation as an universal nuclear transcriptional factor. Bcl-xL and Bak were expressed in cytoplasm of malignant cells in 62 and 20 cancers, respectively. STAT3 correlated both with Bcl-xL (p = 0.001, r = 0.365) and Bak (p  < 0.001, r = 0.436) in all of endometrioid adenocarcinomas and variably in different subgroups of these tumours segregated in regard to grading, staging and patients' age. Remarkably, only ER-α positive cancers retained these correlations in opposition to ER-α negative tumours with negativity defined as an immunoreactivity below 10%. ER-α receptor probably enhances interactions between STAT3 and Bcl-xL to be present in statistically significant manner. Presence of ER-α receptor seems to be crucial for relationships among Bcl-xL and STAT3 to occur in endometrioid adenocarcinomas.

Tumor-infiltrating lymphocytes in tissue material combined with systemic lymphocyte inflammation in patients with colorectal cancer.

The efficacy of cancer immunotherapy has been actively explored in the treatment of various malignant neoplasms of the gastrointestinal tract. In light of recent reports, the present study aimed to investigate the combination of the absolute lymphocyte count (ALC), percentage of tumor-infiltrating lymphocyte (TILs) and tumor progression status in patients with colorectal cancer (CRC) who underwent surgery. The medical records of 160 patients diagnosed with CRC were retrospectively reviewed. TILs were determined as a percentage of mononuclear inflammatory cells in the total intratumoral or stromal area as determined in five high power fields (magnification, x200-400), at the invasive front and in the centre of the tumour. Blood samples were obtained within 3 days prior to and 7 days following the surgical treatment. The assessment of the TIL percentage was performed in the tissue at the invasive front and in the centre of the primary tumour mass in combination with the determination of ALC in whole blood samples. The samples were obtained prior to and after surgery from patients with CRC, and the tumour progression status was also assessed (TILs/ALC/tumour progression status). A significant association was observed between the percentage of TILs in the main mass of tumour and the tumour size (P=0.031), the pT stage (P=0.049) and the incidence of necrosis (P=0.037) following surgery. The histological type was associated with the evaluated combined parameters prior to surgery (P=0.046). Lymph node pouch invasion was associated with TILs at the invasive front of tumour and with ALC prior to and after surgery (P=0.006 and P=0.037). Furthermore, the data indicated that the percentage of TILs located on the invasive front and centre of the tumour, and the ALC prior to and following surgery correlated with the treatment status (P=0.032, P=0.018, P≤0.001 and P≤0.001). A significant association was noted between eight features and evaluated combined parameters following surgery. These included the tumour size (P=0.021), TNM stage (P<0.001), tumour deposits (P=0.001), incidence of necrosis (P=0.042) and lymph node metastasis (P<0.001). Furthermore, the degree of invasion of venous (P<0.001), lymphatic (P<0.001) and perineural (P<0.001) sites was also significantly associated with TILs, ALC obtained after surgical treatment and tumor progression status. The data demonstrated that local and systemic chronic inflammation was associated with tumour progression in patients with CRC.

Monocyte-to-lymphocyte ratio as a prognostic factor in peripheral whole blood samples of colorectal cancer patients.

BACKGROUND: Colorectal cancer is the third most common malignancy worldwide. Therefore, it is critically important to identify new useful markers that can be easily obtained in routine practice. Inflammation is a crucial issue in the pathogenesis and development of cancer. AIM: To evaluate the prognostic value of absolute monocyte count, monocyte to lymphocyte ratio (MLR), the combination of neutrophil-to-lymphocyte ratio and platelet-to-lymphocyte ratio (NLR-PLR), and combined platelet and neutrophil-to-lymphocyte ratio (PLT-NLR) in peripheral blood samples of patients with colorectal cancer undergoing surgery. METHODS: We conducted a retrospective study of 160 patients with colorectal cancer who underwent surgery, and 42 healthy controls. The status of absolute monocyte count, MLR, NLR-PLR and PLT-NLR was calculated on the basis of blood samples obtained before and after surgery. Haematologic factors were examined in correlation with the type of tumour growth, tumour size, histological type, percentage of mucinous component, grade of malignancy, Tumour-Node-Metastasis stage, venous, lymphatic and perineural invasion of cancer cells, status of lymph node invasion and the presence of cancer cell deposits. The Kaplan-Meier method and the long-rank test were used to compare survival curves. To determine independent prognostic factors, univariate and multivariate Cox proportional hazards regression models were applied. RESULTS: The PLT-NLR status was correlated with tumour size and the presence of perineural invasion (P = 0.015; P = -0.174, P = 0.037). Moreover, high NLR-PLR and PLR-NLR ratios in the blood samples obtained after surgery were positively associated with histological type of cancer and percentage of the mucinous component (NLR-PLR: P = 0.002; P = 0.009; PLR-NLR status: P = 0.002; P = 0.007). The analysis of 5-year disease-free survival showed that the MLR of whole blood obtained after surgery [HR = 2.903, 95%CI: (1.368-6.158), P = 0.005] and the status of lymph node metastasis [HR = 0.813, 95%CI: (0.653-1.013), P = 0.050] were independent prognostic factors in colorectal cancer patients. CONCLUSION: The postoperative MLR in whole blood samples can be used as an independent prognostic factor in patients diagnosed with colorectal cancer.

Pre- and postoperative neutrophil and lymphocyte count and neutrophil-to-lymphocyte ratio in patients with colorectal cancer.

Colorectal cancer (CRC) is one of the most common malignant cancers worldwide. Patients with CRC are diagnosed based on various predictors, including performance status, clinicopathological factors and TNM classification. The aim of the present study was to analyze the neutrophil and lymphocyte counts, as well as the neutrophil-to-lymphocyte ratio (NLR) in pre- and postoperative blood samples of patients with CRC in correlation with specific anatomical variables and disease- free survival (DFS). The variables pre- and postoperative neutrophil count (preNEU and postNEU, respectively), lymphocyte count and NLR were significantly higher in cancer patients than those noted in healthy subjects (all P<0.001). PreNEU count correlated with tumor size, necrosis and tumor budding (R=0.204, P=0.014; R=0.189, P=0.023; R=-0.174, P=0.036, respectively). Moreover, postNEU was associated only with the histological type (R=0.174; P=0.047). The PreLYMPH count was correlated with distant metastasis (R=-0.153, P=0.046). PreNLR and postNLR were associated with the expression of various histological markers of disease progression. Analysis of DFS indicated that the postNEU count in the low group exhibited a tendency to lower DFS duration, although the results were not significant (P=0.055). In conclusion, the present study indicated a significant correlation between the factors analyzed in blood samples of CRC patients and the disease progression markers.

Hypoxia related growth factors and p53 in preoperative sera from patients with colorectal cancer--evaluation of the prognostic significance of these agents.

BACKGROUND: Insulin-like growth factor-I (IGF-I) and vascular endothelial growth factor (VEGF) belong to a group of hypoxia related proteins. IGF-I induces expression of VEGF and decomposes wild type p53 in cancer cell lines. The goal of our study was to evaluate serum IGF-I, VEGF and p53 with respect to overall and disease free survival of patients with colorectal cancer (CRC) patients compared with healthy volunteers. METHODS: Preoperative blood samples from 125 patients with CRC and 16 healthy volunteers were examined using ELISA for serum IGF-I, p53 and VEGF concentrations. RESULTS: Concentrations of p53 and VEGF were significantly higher in CRC patients than in controls (p<0.0006 and p<0.0001, respectively). IGF-I was not statistically different between both groups. Serum IGF-I showed negative correlation with p53 in CRC patients (p<0.04, r=-0.193). IGF-I and VEGF showed negative correlation in poorly differentiated cancers (G3) (p<0.03, r=-0.339). Patients with VEGF concentrations that were above average for the cancer population survived for a shorter period of time (p=0.065 in evaluation of overall survival and 0.071 in estimation of disease-free survival during a 3-year follow-up) compared with patients with serum VEGF lower than the highest values seen in controls. CONCLUSIONS: Comparisons between serum IGF-I and p53 appear to confirm the metabolism of p53 by IGF-I. Serum VEGF showed prognostic significance in our study. Serum concentrations of IGF-I and VEGF did not show positive correlation, as expected due to IGF-I induction of VEGF in malignant colon cell lines.

Prognostic significance of inflammatory cell response in patients with colorectal cancer.

Cancer cells are characterized by a low antigenic immunogenicity, a rapid growth and an immunosuppressive effect on the extracellular matrix. These properties induce a weak immune response in colorectal cancer (CRC) carcinogenesis. It is therefore crucial to determine the composition of the inflammatory mass, including neutrophils, macrophages and eosinophils in the tumor tissue of patients with CRC, and to analyze other clinicopathological parameters. The present study included 144 patients diagnosed with CRC. Tissue samples obtained from routine histopathological diagnosis were stained with hematoxylin and eosin. Inflammatory cells were assessed in the invasive front and in the center of the tumor by light microscopy under a high-power magnification. The percentage of neutrophils in the invasive front was significantly higher compared with that in the center of the tumor mass (P<0.01). Macrophages and eosinophils were present in the invasive front and in the center of tumor mass in most cases. The presence of neutrophils, macrophages and eosinophils was correlated with various clinicopathological features. Patients with macrophages present in the center of tumor mass had longer disease-free survival time (P=0.041). In conclusion, the present study demonstrated that the inflammatory cell infiltrate served a significant role in the immune response of patients with CRC. It should be noted that the presence of macrophages localized in the stroma of the central part of the primary tumor mass was associated with the survival time of patients with CRC.

Tumor-infiltrating lymphocytes in primary tumors of colorectal cancer and their metastases.

The presence of tumor cells in the large intestine stimulates hypoxia and local inflammatory mediators that activate numerous inflammatory cells, including a diverse lymphoid tumor cell population. The aim of the present study was to evaluate tumor-infiltrating lymphocytes (TILs) located in the invasive primary tumor, surrounding deposits of tumor cells and those present in distal metastatic cells in the liver of patients with colorectal cancer. Furthermore, the correlation of TILs with anatomical parameters was assessed. The study group included 123 patients with primary tumor colorectal cancer without distant metastasis, 25 cases with deposits of colorectal cancer cells and 15 cases of colorectal cancer liver metastasis. TILs were assessed in tissues stained with hematoxylin-eosin using light microscopy and evaluated by two independent pathologists blinded to the clinical information. Infiltration of TILs in the invasive front of primary tumor was stronger compared with those surrounding deposits of cancer cells and liver metastases (P<0.001). TILs in the invasive front of primary tumor masses were associated with various variables linked with tumor progression and inflammatory cell infiltrate. TILs distributed around the deposits of cancer cells were associated with postoperative treatment; however, those localized in the invasive front of liver metastases were correlated with preoperative therapy. In conclusion, TILs assessment in primary tumors of colorectal cancer, surrounding deposits of tumor cells and in the metastatic cells in the liver may be helpful in understanding the role of these cells in the organization of immune response.

Squamous cell carcinoma of the breast as a clinical diagnostic challenge.

Squamous cell carcinoma (SqCC) of the breast should be differentiated between the primary skin keratinizing squamous carcinoma and squamous metaplastic cancer. In the current study, the cases of two patients who were diagnosed with SqCC originated from skin and the breast were discussed. A fine-needle aspiration biopsy confirmed the presence of atypical squamous cells. In both cases, the microscopic examination of the surgical specimen revealed a malignant neoplasm differentiated into SqCC characterized by keratinizing cancer cells with abundant eosiphilic cytoplasm with large, hyperchromatic vesicular nuclei. Immunohistochemical studies showed negative for progesterone and estrogen receptors and human epidermal growth factor receptor 2. Moreover, negative expression of cytokeratin 7 and 20 was confirmed. The diagnosis of the both tumors was established based on the detailed analysis of clinical, macroscopical and microscopical information. SqCC localized in the breast is a great diagnostic challenge in pathomorphology and more attention should be paid for analysis of such lesions in daily practice.

STAT1 and STAT3 as intracellular regulators of vascular remodeling.

The roles of signal transducers and activators of transcription (STAT) proteins are widely discussed in the pathogenesis of cardiovascular diseases. It is highly probable that STAT1 and STAT3 are activated during proliferation and inflammation inside atheromatous plaques. Luminal surfaces of endothelium become thrombogenic because of STAT1-dependent induction of MHC II and STAT3-regulated recruitment of phospholipase A2. As with STAT1, STAT3 seems to mediate stimulation of vascular wall cells by VEGF, HGF, and Ang II. STAT3 can contribute to counteracting apoptosis by eventual cooperation with c-fos and the bcl-xl gene. As pharmacological agents called statins are reported to regulate activities of STAT proteins, these signal messenger proteins could serve as targets for anti-atherogenic therapy. We attempted to review the role of STAT1 and STAT3 proteins in vascular remodeling.

Diagnostic value of inflammatory cell infiltrates, tumor stroma percentage and disease-free survival in patients with colorectal cancer.

The anticancer immune defense mechanism involves humoral and cellular responses. The main effector mechanisms of antitumor responses involve the following: the activity of cytotoxic T cells; the activation of macrophages and neutrophils; the activity of cytokines secreted by T cells; and natural killer cell activity. Selected cell populations are responsible for the stimulation or suppression of the immune system against tumor cells. Therefore, the aim of the present study was to evaluate the location, extent and composition of the cellular inflammatory infiltration of tumors in patients with colorectal cancer (CRC). In addition, the correlation between cellular inflammatory infiltration, and anatomoclinical and histopathological features of patients was evaluated. The study involved 160 patients diagnosed with primary operable CRC. The local inflammatory infiltrate was assessed in the invasive front and center of the tumor using light microscopy with hematoxylin and eosin (H&E) staining, according to the Klintrup-Makinen criteria, tumor stroma percentage, and Glasgow microenvironment score. The inflammatory infiltrate in the invasive front of the tumor was correlated with gender (P=0.018), the invasion of blood vessels (P=0.020) and lymph vessels (P=0.038), the presence of tumor-infiltrating lymphocytes in the invasive front (P=0.033) and center (P<0.001) of the tumor, fibrosis (P<0.001), and the degree of desmoplasmic stroma (P=0.004). In contrast, inflammatory infiltration in the center of the tumor was associated with the tumor node metastasis stage (P=0.012), Dukes' stage (P=0.009), primary tumor stage (P=0.036), lymph node status (P=0.005), number of lymph nodes (P=0.006), invasion of lymph node pouches (P=0.021), size of lymph node metastasis (P=0.025) and the degree of desmoplasmic stroma (P=0.002). The low-group, who demonstrated an absent or weak inflammatory cell infiltrate in the invasive front of the tumor, had a statistically significant shorter disease-free survival (DFS) time (P=0.004). Inflammatory cell infiltrate in the invasive front was identified as an independent predictive factor in CRC (P=0.041). In conclusion, the degree of inflammatory cell infiltration in the invasive front of the primary tumor significantly affects various variables that determine disease progression and DFS rates of patients with CRC. Furthermore, the routine histopathological assessment of this parameter in tissue stained with H&E may have potential prognostic value.

Stromal and intraepithelial tumor-infiltrating lymphocytes in colorectal carcinoma.

The local mechanisms of antitumor immune defense determine the development and organization of the tumor microenvironment, and the composition and relative proportions of the inflammatory cell population affect the quality and characteristics of the immune response. The aim of the present study was to conduct a quantitative morphological evaluation of two types of tumor-infiltrating lymphocyte (TILs) populations, including those located in the stroma and intraepithelial cancer structures, in the invasive front and the center of the tumor in patients with colorectal cancer (CRC). The study included 160 patients with CRC who had undergone surgery. The tissue material was stained with hematoxylin and eosin, as used in routine histopathological diagnosis, and the two TIL populations were observed and counted with light microscopy. The relative extent of infiltration of stromal and intraepithelial TILs into the front and center of the primary tumors was similar. The extent of infiltration by stromal TILs was negatively correlated with the morphological features of tumor progression including the cancer infiltration of blood vessels (P=0.016), the invasion of lymph vessels (P=0.007), perineural invasion (P=0.036), lymph node involvement (P=0.047) and distant metastases (P=0.032). The infiltration by intraepithelial TILs was positively correlated with a desmoplastic reaction (P=0.002). Disease-free survival time was statistically shorter in patients without intraepithelial TILs in the center of the primary tumor mass (P=0.049; hazard ratio = 1.45). These results confirm that the infiltration of TILs into the invasive front and center of the tumor in patients with CRC serves an important role in the invasion and progression of the disease, and should be considered in routine histopathological examinations.

Levels of VE-cadherin increase independently of VEGF in preoperative sera of patients with colorectal cancer.

AIMS AND BACKGROUND: Vascular endothelial cadherin (VE-cadherin) preserves the tightness of the mature vascular network as a component of endothelial adherens junctions. Vascular endothelial growth factor (VEGF) makes VE-cadherin dissociate from complexes with beta-catenin, so that endothelial cells can loosely proliferate and migrate. We searched for relationships between VEGF and VE-cadherin levels in preoperative sera of patients with colorectal cancer (CRC). We also compared VE-cadherin levels of control and preoperative CRC sera in relation to clinicopathological features. METHODS: We measured with an ELISA kit the serum levels of the proteins in preoperative samples from 125 CRC patients and in samples from 16 healthy volunteers. RESULTS: Serum VE-cadherin was about fourfold higher in CRC patients than in controls (P < 0.00001), with similar results being found in subgroups with different clinicopathological features versus controls. VE-cadherin was not correlated with VEGF in the entire group of CRC patients nor in the subgroups of node-positive and node-negative patients, different grades of histological differentiation (G2 or G3), extent of tumor growth (pT1+pT2 or pT3+pT4), histopathological type (adenocarcinoma or mucinous carcinoma), sex, age, and tumor site (colon or rectum). However, the serum levels of VE-cadherin and VEGF in CRC patients, which were higher than the mean values of controls, tended towards a negative correlation in node-positive patients (P = 0.078, r = -0.279). CONCLUSIONS: VEGF and VE-cadherin seem to be independent markers of angiogenesis in CRC with no significant correlation between their serum levels.

Immunohistochemical expression and serum level of survivin protein in colorectal cancer patients.

Survivin is one of the apoptosis-related inhibitors that is associated with a more aggressive behavior and a poor prognosis in numerous types of malignancies, including colorectal cancer (CRC). The objective of the present study was to perform immunohistochemical tissue analysis of survivin expression and serum analysis of survivin levels in CRC patients. The study group consisted of 55 CRC patients. Survivin expression was assessed by immunohistochemistry in 38 patients using monoclonal antibodies. Color reactions were observed in the nucleus and cytoplasm of the cancer cells. The expression was defined based on the H-score method. The level of survivin was determined using the enzyme-linked immunosorbent assay method. A positive immunoreaction was observed in the tumor tissues of 84.2% (32/38) of patients with CRC, consisting of nuclear (63.2%; 24/38) and cytoplasmic (81.6%; 31/38) expression. The survivin nuclear expression was associated with tumor mass location and the presence of distant metastases (P=0.048 and P=0.026, respectively). Survivin was detected in the sera of 38.2% (21/55) of CRC patients and in 81.8% (18/22) of healthy individuals. Serum protein levels were found to correlate with hematocrit (P=0.035), hemoglobin (P=0.008) and albumin (P=0.045), but not with any of the investigated clinicopathological parameters. The immunohistochemical positive reaction of survivin in the nuclei of cancer cells may condition their proliferative capacity, which is associated with higher risk of developing metastatic foci. Thus, the present study suggests that the expression of survivin may have diagnostic implications in cancer of the colon and thus requires further research. By contrast, the survivin serum level in CRC patients appears to be diagnostically ineffectual for clinical use.

Reduced expression of caspase-8 and cleaved caspase-3 in pancreatic ductal adenocarcinoma cells.

Pancreatic ductal adenocarcinoma (PDAC) is a rare neoplasm that affects the gastrointestinal system, and is characterized by a high mortality rate. It has been demonstrated that apoptosis has a significant role in the regulation of cancer cells. Therefore, the aim of the present study was to immunohistochemically assess the expression of proteins belonging to the caspase family, namely caspase-8, pro-caspase-3 and cleaved (active) caspase-3 in pancreatic cancer. The study group consisted of 29 patients exhibiting PDAC. Protein expression was evaluated by immunohistochemical methods. The expression of caspase-8 in normal cells was negative in 17.2% of cases and positive in 82.8% of cases. All cases demonstrated pro-caspase-3 expression in normal pancreatic cells, compared with 93.1% of cancer cells. Staining for activated caspase-3 was positive in 27 normal tissue cases, compared with positivity in only 10 cancer cases. Caspase-8 expression positively correlated with cleaved caspase-3 expression in the cytoplasm of cancer cells (P<0.002). Caspase-3 expression was identified to correlate with inflammatory peritumoral infiltration (P<0.015). No correlation was observed between caspase expression and any other clinicopathological parameters. The results of the present study demonstrated aberrant initiation of cancer cell apoptosis in PDAC via a decrease in caspase-8 expression, which may lead to disorders in the activation of effector caspase-3.

Tumour 'budding' and its relationship to p53 and Bcl-2 expression in colorectal cancer.

The formation of poorly-differentiated clusters of cancer cells in the tumour growth zone, defined as "tumour budding", is a likely factor determining the biological malignancy of colorectal cancer (CRC). The aim of the study was to evaluate tumour budding in the CRC growth zone and to analyse its relationship to chosen anatomoclinical parameters, and to p53 and Bcl-2 protein expression in the tumour budding and in the main tumour mass. Fifty-seven colorectal cancers, classified as pT3 and G2, were used for analysis. Immunohistochemical investigations were performed using the anti-human p53 and Bcl-2 protein monoclonal antibodies (Dako/p53, No M7001 and Dako/Bcl-2, No M 0887, respectively). It has been found that p53 overexpression in the primary tumour and the presence of lymph node metastases correlate with strongly-positive tumour budding (p < 0.04 and p < 0.000001). However, low expression of p53 protein in the primary tumour and lack of lymph node metastases was statistically significantly correlated with the absence of tumour budding. A statistically significant correlation was shown between p53 protein expression in the tumour budding and expression of p53 in the primary tumour and lymph node metastases (p < 0.05). We also observed a statistically significant correlation between Bcl-2 protein expression in the tumour budding and its expression in the primary tumour and lymph node metastases (p < 0.05). These data suggest that tumour budding of cancer, in combination with other markers, may provide more information about the biological behaviour of colorectal cancer.

The expression of tumorigenesis markers in oral papilloma.

Oral papilloma is the most frequent benign tumor of the oral cavity but its biological potential for malignant transformation is still to be evaluated. The alteration of apoptosis and uncontrolled cell proliferation is considered to be an important factor in oral tumorigenesis. The aim of our study was to evaluate by immunohistochemistry P53, Bcl-2, CD44 and PCNA expression in oral papillomas with and without dysplasia. We examined a series of 55 oral papillomas, including 12(21.8%) cases of papillomas with epithelial dysplasia. Staining patterns were correlated with sex, age, tumor location, size and presence or absence of epithelial dysplasia. P53 showed positive reaction in 70.9%, PCNA in 80%, CD44 in 50.9% and Bcl-2 in 21.8% of papillomas. There was no correlation between sex, age, tumor size, location and presence of dysplastic epithelium in papillomas. We observed a statistically significant correlation between Bcl-2, CD44 expression and presence of epithelial dysplasia in papillomas. Coexistence of PCNA and P53 positive immunostaining was observed. Papillomas with overexpression of P53 and PCNA showed negative reaction for CD44 protein. The results of our study suggest that overexpression of P53 and PCNA might be an early event in oral tumorigenesis, whereas CD44 and Bcl-2 are potential markers of epithelial dysplasia in oral papillomas.

Correlation between Bcl-2 protein expression and some clinicopathological features of oral squamous cell carcinoma.

Bcl-2 inhibits most kinds of programmed cell death and provides a selective survival advantage to various cell types. The biological significance of Bcl-2 expression for the development and progression of oral cancer has still to be evaluated. The aim of our study was to estimate possible correlations between the Bcl-2 protein expression and some clinicopathological features of oral cancer. The study was conduced on 129 patients treated surgically for oral squamous cell carcinoma. The statistically significant relationships were observed between oral squamous cell cancer Bcl-2 expression and higher tumor grading (p<0.005), higher tumor mitotic index (p<0.005), higher index of atypical mitoses (p<0.001) as well as microfocal pattern of tumor invasive margin (p<0.001). The results suggest that positive Bcl-2 expression may be a valuable factor supplementing the established unfavorable histopathological features of oral squamous cell carcinoma.

Green tea as an antioxidant which protects against alcohol induced injury in rats -- a histopathological examination.

Our study with animal models was designed to test the hypothesis that green tea protects against chronic (over 4 weeks) alcohol induced liver injury in rats. The research was conducted on Wistar male rats divided into 4 research groups: I - received the Libera-De Carli control diet (L-DC), II - received (L-DC) and green tea, III - received (L-DC) and ethanol and IV - received (L-DC), green tea and ethanol. When comparing groups I and II we saw less intensive steatosis in group II than in group I, which can suggest that green tea may affect the accumulation of fat in the hepatocytes and protect them against steatosis and disruption. In III, the ethanol group, the steatosis of the liver increased considerably and the green tea which was given with ethanol in group IV did not halt this, as in group IV we also observed intensive steatosis in the liver. From this data we conclude that green tea has an important, although not fully understood role in preventing liver injury.

Expression of connexin 43 in breast cancer in comparison with mammary dysplasia and the normal mammary gland.

Gap junctional intercellular communication (GJIC) plays a critical role in tissue development and differentiation and probably in carcinogenesis. The purpose of the study was to evaluate the expression and localisation of Cx43 in 40 cases of mammary dysplasia and 29 cases of breast cancer (without primary chemotherapy). The tissue sections were investigated for Cx43 expression by immunohistochemistry. In the normal mammary gland there was an intercellular, punctate staining pattern, mainly between myoepithelial cells, characteristic of functional gap junctions. In dysplasias there was mainly mixed (cytoplasmic and intercellular) staining and in most cases of breast cancer we observed diffuse or granular, but cytoplasmic, staining of Cx43. Our results demonstrated that expression of Cx43 in dysplasias and breast cancer is changed and GJIC is probably impaired because of disruption of functional gap junction formation especially between breast cancer cells.