Protective Role of IL7A-197 A/G Heterozygosity in the Development and Severity of Colorectal Cancer in the Bulgarian Population.

Background: Interleukin (IL)-17A and IL-17F, expressed mainly by a novel subset of CD-positive (+) T-helper (Th) cells of the immune system, has been closely related to inflammatory conditions underlying colorectal cancer pathogenesis. Accordingly, we conducted a case-control study to investigate the association of common single nucleotide polymorphisms (SNP) in the IL17A and IL17F genes (rs2275913 and rs763780, respectively) with the susceptibility and severity of CRC patients from the Bulgarian population. Methods and Materials: 136 patients with histologically confirmed CRC diagnosis and 116 healthy individuals were recruited in the present study. Genotypes were determined by the restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) technique. Results: The IL17A heterozygous A/G-genotype was overrepresented among the control group (p = 0.003). Additionally, the carriers of the heterozygous A/G-genotype had a 2.39-fold lower risk for CRC compared to the G/G-genotype (OR = 0.418, p = 0.006). Our results also indicated that in the advanced CRC stages (III + IV) the heterozygous genotype (A/G) appeared to be less frequent (p = 0.024, χ2-test). Among the patients with detected distant metastases, the A/G-carriers were the smallest part (14.3%) compared to the homozygous genotypes A/A (42.9%) and G/G (42.8%), p = 0.006. There was no association of the studied IL17F rs763780 SNP with susceptibility and severity of CRC among the studied subjects, although the heterozygous C/T-carriers had shorter median survival compared to the T/T-carriers (p = 0.129). Conclusions: Our study finds a protective role of heterozygosity for the IL17A-197A/G SNP and negative effects of the A-allele on CRC progression.

CD11c- and CD123-positive dendritic cells in development of antitumour immunity in non-small cell lung cancer patients.

Our aim was to analyzed the significance of CD11c and CD123 positive DCs and their relations with some clinical and pathologic parameters of patients with non-small cell lung cancer (NSCLC). The immunohistochemical expression of CD11c and CD123, was evaluated in 40 patients with NSCLC. After analysis we found that 35.3% of the patients in the T3-4 tumour stage had a high CD11c infiltration in the tumour stroma, while 100% of the patients in the T1-2 tumour stage had low infiltration (p = 0.03). We also found that 71.4% of patients in the M1 stage had a high infiltration with CD123 in the tumour stroma, whereas only 15.6% of patients without metastases had high infiltration, analogous data are also found in comparing the distribution of CD123 in the tumour border (p = 0.002 or p = 0.002). Comparing the density of CD123 in the border of lymph node involvement, we found that only 7.14% of patients without metastases had low infiltration with dendritic cells, whereas in patients with metastatic lymph nodes that percentage was 41.7% (p = 0.008). In conclusion results suggest that CD11c- and CD123-positive DCs play an important role in antitumour immunity and can be predictive factor for tumour development in patients with NSCLC.

Promoter polymorphisms in TGFB1 and IL10 genes influence tumor dendritic cells infiltration, development and prognosis of colorectal cancer.

BACKGROUND: Anti-inflammatory cytokines such as interleukin (IL)-10 and transforming growth factor (TGF)-ß1 have a complex role in the development of colorectal cancer (CRC). Dendritic cells (DCs) are the cellular component of the inflammatory microenvironment in the tumor and infiltration of tumors by DCs is associated with better prognosis and fewer metastases. METHODS: In the present study, we explored the role of two single nucleotide polymorphisms (SNPs) in the promoter regions of TGFB1 and IL10 genes and their associations with infiltrating DCs in CRC.A case-control study was designed. Genotyping was performed via the polymerase chain reaction-restriction fragment length polymorphism method and DC infiltration was determined immunohistochemically. RESULTS: For the TGFΒ1 -509C/T SNP, we found that the T allele was less frequent in patients than in controls (p = 0.031) and the TT-genotype had a 2.74-fold lower risk for CRC than the CC-genotype (odds ratio = 0.365, 95% confidence interval = 0.15-0.88, p = 0.015). Additionally, the TT carriers had the shortest median survival (14.4 months) (p = 0.045). The C-allele genotypes had a significantly longer survival compared to TT carriers (p = 0.018). The CC genotype was associated with a lower cellular density of CD11c in the invasive margin of the tumor (p = 0.033), whereas there was an opposite finding for CD83+ DCs (p = 0.037). Carriers of A-allele genotypes of the IL10 -1080A/G SNP had significantly lower CD83+ cells (p = 0.046) in the tumor invasive margin. CONCLUSIONS: The T-allele of the TGFB1 -509C/T SNP might be a protective factor for development of CRC, although, in the course of the disease, this variant allele might be associated with more unfavorable prognosis of the patients.

Impact of TGF-ß1 expression and -509C>T polymorphism in the TGF-ß1 gene on the progression and survival of gastric cancer.

The aim of this study was to examine the expression of TGF-ß1 and TGF-ß receptor type II (RII) and the impact of the -509C>T single nucleotide polymorphism (SNP) in the gene in relation to clinicopathological factors in gastric cancer (GC). Using immunohistochemistry we investigated 43 patients with GC for expression of TGF-ß1 and TGF-ß-RII. Consequently, RFLP-PCR was performed to analyze the presence of -509C>T polymorphism in the TGF-ß1 gene. We found that 72.1% of GCs had cytoplasmic TGF-ß1 expression and 27.9% were negative. The TGF-ß1 receptor type II was expressed on tumor cell membranes in 58.1%. TGF-ß1 positivity in tumor cytoplasm correlated positively with TGF-ß1-RII expression in tumor cytoplasm in 67.4% of cases (2 = 8.02; p = 0.005). Also, the results showed that patients with low and moderate tumor differentiation had TGF-ß1-RII positivity in 53.3% and 81.8% resp. (2 = 6.58; p = 0,037). The analysis of genotype distribution of the -509C>T SNP in the promoter region of TGF-ß1 gene and clinical stage distribution revealed that among the 32 patients in III-IV clinical stage 53.1% were heterozygous (TC), 34.4% were homozygous for the C-allele and 12.5% were homozygous for the variant T-allele (2 = 3.31; p = 0.069). In conclusion the expression of TGF-ß1 was related to shorter survival time and rapid progression for the GC patients. Additionally, the variant T-allele of the studied polymorphism was associated with worse prognosis for GC patients.

Association of IL-12Bpro polymorphism with tumor-infiltrating dendritic cells in colorectal cancer.

PURPOSE: Chronic inflammation is a key component in the development and progression of colorectal cancer (CRC). A notable hallmark of the inflammation process is the release of pro-inflammatory cytokines by infiltrating cells of the immune system. Defects in dendritic cells (DCs) recruitment, maturation and cytokine release are a hallmark of the CRC strategy to escape immune surveillance.The purpose of our study was to evaluate the possible role of IL-12B polymorphism in the promoter region of the IL-12B gene (rs17860508) as a genetic factor contributing to the risk for CRC development. Additionally, we aimed to evaluate the influence of this polymorphism on DCs infiltration in tumor microenvironment. METHODS: IL-12Bpro polymorphism was genotyped by Amplification Refractory Mutation System- Polymerase Chain Reaction (ARMS-PCR). Immunohistochemistry was performed for DCs infiltration. RESULTS: Statistically significant correlation between the expression of S100 and CD1a DCs and the 11- genotype of the studied polymorphism was found. No statistically significant difference in genotype distribution between cases and controls was observed (p=0.163). Analysis of the overall survival (OS) of genotyped patients revealed a tendency among the carriers of the 22-genotype to have shorter survival of 36 months versus the 11- and 12-cariers- 61 months (log rank, p=0.117). CONCLUSIONS: The IL-12Bpro polymorphism does not constitute a risk factor for CRC development. However, genotype-11 might have a complex role in the recruitment and maturation of DCs in tumor microenvironment.

Impact of HER2 codon 655 polymorphism and expression of HER2 and HER3 in non small cell lung cancer patients.

The aim of the study was to assess the expression and significance of HER2 and HER3, and Ile/Val single nucleotide polymorphism (SNP) of HER2 in lung cancer patients. Thirty seven cases of lung cancer were investigated immunohistochemically for HER2 and HER3 expression. PCR followed by restriction fragment length polymorphism (RFLP) was used to analyze the presence of HER-2 SNP at codon 655 in 20 samples. The results were compared with clinical and pathological parameters of investigated patients.We found that 100% of the cases were negative for HER2, 29.7% were with moderate or strong HER3 expression and 70.3% of the tumors-without or with low expression for HER3. Lymph node metastasis were found in 40% of HER3 positive cases (χ(2) = 4.752; p = 0.029). Moderately-differentiated tumors do not express neither of investigated markers (χ(2) = 6.719; p = 0.035). HER2 RFLP-PCR analysis showed genotype AG in five patients (25%) and the rest of 15 cases (75%) had АА (Ile/Ile) genotype. Patients with metastasis had genotype АА (Ile/Ile) in 80% and genotype AG (Ile/Val) in 20% (χ(2) = 2.857; p = 0.091).Our results indicate that SNP in HER2 codon 655 and investigation of HER2 and HER3 expression could be helpful to outline the prognosis for patients with lung adenocarcinoma.

VEGF expression, microvessel density and dendritic cell decrease in thyroid cancer.

Thyroid cancer is one of the five most common cancers in the age between 20 and 50 years. Many factors including the potent angiogenic vascular endothelial growth factor (VEGF) and different dendritic cell types are known to be related to thyroid tumourogenesis. The study was performed to address the expression of VEGF and microvessel density in thyroid cancers and to evaluate the effect of VEGF expression in thyroid tumour cells on the dendritic cells. We investigated 65 patients with different types of thyroid carcinomas: papillary (PTC), oncocytic (OTC), follicular (FTC) and anaplastic (ATC), immunohistochemically with antibodies against VEGF, CD1a, CD83, S100 and CD31. Our results suggest that the expression of VEGF is significantly more often in PTC than ATC (92.3% vs. 60.0%, p = 0.025). The microvessel density marked with CD31 in the tumour border of PTC was significantly higher as compared to FTC (p = 0.039), but not to ATC and OTC (p = 0.337 and 0.134). We found that CD1a- and CD83-positive cells were dispersed with variable density and in OC CD31+ vessel numbers were positively correlated with CD83+ dendritic cells in tumour stroma (R = 0.847, p = 0.016). We did not find statistically significant associations of the survival of patients with PTC after the surgical therapy with VEGF expression and MVD. In conclusion we may state that VEGF expression in tumour cells of thyroid cancer can induce neovascularization and suppress dendritic cells.

Investigation of the role of MMP3 -1171insA polymorphism in cutaneous malignant melanoma - a preliminary study.

Coetaneous malignant melanoma is the most aggressive cancer of the skin with a high rate of mortality worldwide. Degradation of basement membranes and extracellular matrix is an essential step in cancer invasion and metastasis. Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) play key roles in this step. MMP-3 also called stromelysin-1 was one of the first proteinases found to be associated with cancer. In the gene of MMP-3 (MMP3), an insertion/deletion of an A nucleotide at position -1171 in promoter region has been identified and shown to effect the expression activity of the gene. The present study was conducted to investigate the relation of MMP3 -1171insA polymorphism with skin malignant melanoma risk in a pilot case-control study of Bulgarian patients (n = 26) and unaffected controls (n = 172). The genotypes of controls and melanoma patients were in Hardy-Weinberg equilibrium. The results showed no statistically significant difference both in genotype and allele frequencies of MMP3 -1171insA polymorphism between melanoma patients and healthy controls either in crude analyses (p = 0.360 and 0.790, c2-test) or after adjustment for age and sex. The comparison of some clinical characteristics between the patients with different genotypes showed a trend for longer survival of patients with 6A/6A genotype compared to the carriers of 5A allele (5A/5A+5A/6A genotypes, p = 0.118, Log rank test). The results of our current preliminary study do not provide evidence for the role of the promoter polymorphism -1171insA in MMP3 as a risk factor for development of coetaneous melanoma, but suggest its implication in progression of the diseases.

Interleukin-6-Positive Immune Cells as a Possible New Immunologic Marker Associated With the Colorectal Cancer Prognosis.

Chronic inflammation creates tumor microenvironment (TME) that facilitates colorectal cancer (CRC) cell proliferation, migration, metastasis, and tumor progression. Interleukin-6 (IL-6) is a proinflammatory cytokine with a pleiotropic effect on CRC development. We aimed to evaluate IL-6 expression in tumor cells and in immune cells in TME, to assess the serum level and IL6 -174 G/C genotype distribution and to correlate the results with selected morphologic and clinical parameters that may add useful information in understanding the mechanisms of human CRC progression. A total of 153 patients with CRC were recruited in the current study. We assessed the IL-6 serum concentration through the ELISA method, the expression of IL-6 in tumor and in immune cells by immunohistochemical and double immunofluorescence staining, the MSI status by immunоhistochemistry for 4 mismatch repair (MMR) proteins, and the genotype distributions for IL6 -174G/C (rs1800795) single-nucleotide polymorphism through PCR-RFLP method. Our results showed that serum IL-6 level were increased in CRC patients as compared with healthy controls (P<0.0001), and in patients with cancers with advanced histologic type (type IV). However, the higher concentration (above the median of 55.71 pg/mL) was with borderline association with longer survival of the patients after surgical therapy (P=0.055, Log rank test). We also found that IL-6+ immune cells prevailed in the invasive front (IF) of tumors compared with the tumor stroma (TS) (P<0.0001). More IL-6+ cells were recruited in the tumors with less advanced histologic type (I+II), with stronger inflammatory infiltrate in the IF, in early pTNM stages (I+II), without lymph node and distant metastases and the higher levels of IL-6+ cells, especially in the IF, were associated with longer survival (P=0.012). The results of our study suggest that although the serum levels of IL-6 are higher in CRC, the increased IL-6+ cells in tumor microenvironment, both in the invasive front and in tumor stroma, as well as the higher serum levels are associated with good prognostic variables and longer survival of the patients mainly in the early stages of CRC.

Onychomycosis: modern diagnostic and treatment approaches.

The medical term onychomycosis should be understood as chronic infection of the nails caused by a fungus. The most common causative agents are the dermatophytes and Candida species. The less common are certain types of moulds (nondermatophyte moulds or NDMs). In approximately 60-80 % of the cases, onychomycosis is due to dermatophytes. Among dermatophytes, the most often isolated causative pathogen is Trichophyton (T.) rubrum. Other common species are T. interdigitale (formerly T. mentagrophytes), Epidermophyton floccosum, and T. tonsurans. The most significant yeasts causing onychomycosis are Candida albicans and Candida parapsilosis. Predisposing factors for onychomycosis include mainly diseases such as diabetes mellitus, peripheral vascular arterial disease, chronic venous insufficiency, polyneuropathies of diverse etiologies, and immunosuppression, e.g., myeloproliferative diseases (such as lymphoma and paraproteinemia), HIV/AIDS, etc. Other factors facilitating the fungal infection are frequent trauma in professional sportsmen, often accompanied by excessive perspiration. The diagnostic methods that are often applied in different dermatologic departments and ambulatory units are also different. This precludes the creation of a unified diagnostic algorithm that could be used everywhere as a possible standard. In most of the cases, the method of choice depends on the specialist's individual experience. The therapeutic approach depends mostly on the fungal organism identified by the dermatologist or mycologist. This review hereby includes the conventional as well as the newest and most reliable and modern methods used for the identification of the pathogens causing onychomycosis. Moreover, detailed information is suggested, about the choice of therapeutic scheme in case whether dermatophytes, moulds, or yeasts have been identified as causative agents. A thorough discussion of the schemes and duration of the antifungal therapy in certain groups of patients have been included.

Atypical fibroxanthoma-a diagnosis of exclusion!

Fibrohistiocytic tumors of the skin comprise a large range of lesions. One such tumor is the atypical fibroxanthoma (AFX), which is widely considered as a "pseudomalignant" tumor. It is derived from fibroblasts and expresses a variety of histiocytic markers. We present a case of AFX, localized in the right temporal region of the scalp, successfully treated with surgical excision. Immunohistochemical staining helps differentiate this tumor from others in the clinical differential diagnosis, including malignant melanoma, squamous cell carcinoma, and other nonmelanocytic spindle cell tumors such as leiomyosarcoma, rhabdomyosarcoma, angiosarcoma, liposarcoma, and dermatofibrosarcoma protuberans. Historically, AFX was believed to be a superficial variant of malignant fibrous histiocytoma (MFH). However, MFH is now considered a more generalized term for a sarcomatous neoplasm of the subcutaneous tissue. The histopathology of MFH shares features with some malignant mesenchymal neoplasms such as liposarcoma, leiomyosarcoma, rhabdomyosarcoma, and angiosarcoma, but can be differentiated using immunohistochemistry and/or electron microscopy. More recently, the examples of MFH that do not exhibit a more specific line of differentiation have been reclassified as undifferentiated pleomorphic sarcoma (UPS). Many authors currently cannot draw a distinction between AFX and UPS. The clinical and histopathological differences between AFX and UPS are often difficult to delineate. It is probable that they represent two poles of the same disease. Surgical excision in the patient we describe resulted in excellent aesthetic results with lack of recurrence in the 7-month postoperative period.

Development of pancreatic islet cells in the extrahepatic bile ducts of rats with experimentally-induced metabolic syndrome.

CONTEXT: There is data about the existence of some endocrine cells in the epithelial layer of the bile duct in humans and rats. OBJECTIVE: We evaluated Ghrelin-, Insulin-, Glucagon- and Somatostatin-positive cells in peribiliary glands, mast cells, and nerve fibres. MATERIALS AND METHODS: Wistar rats were used for dietary manipulation with a 15% fructose solution for 12 weeks. Tissue samples were elaborated with immunohistochemistry for Insulin, Glucagon, Ghrelin, and Somatostatin. Glucose and lipid parameters were studied. RESULTS: In treated animals, Ghrelin+ and Insulin+ cells in perybiliary glands (PBGs) were significantly increased. In the male fructose group there was a significant increase of the homeostasis model assessment insulin resistance (HOMA-IR). CONCLUSIONS: Stem/progenitor cells in extrahepatic bile tree (EHBT) could be a source of Insulin-producing cells in metabolic syndrome. Fructose treatment induces the increase of Ghrelin+ and Insulin+ cells in PBGs and the elevation of Insulin and Ghrelin plasma concentration.

Glucagon- and insulin-immunopositive endocrine cells in porcine extrahepatic bile ducts and gallbladder.

Introduction: Pancreatic ß-cells and α-cells have been found in the murine extrahepatic biliary ducts but not in the gallbladder. However, there has been no information reported in the specialized literature about the presence of glucagon- and insulin-expressing endocrine cells in porcine bile ducts and gallbladder. Aim: We aimed to perform an immunohistochemical study to identify glucagon- and insulin-positive cells and their distribution in the porcine extrahepatic biliary ducts and gallbladder. Method: The immunohistochemical method was used to detect the presence and distribution of glucagon- and insulin-positive endocrine cells in the common hepatic duct (ductus hepaticus communis), common bile duct (ductus choledochus), cystic duct (ductus cysticus), and gallbladder (vesica fellea) of male pigs. Chromogranin A was used as a typical marker for endocrine cells. Results: The density of chromogranin A-, glucagon- and insulin-positive cells per field was the largest in the common bile duct, followed by the common hepatic duct, cystic duct, and gallbladder. The three types of endocrine cells showed specific localization in the superficial and deep glands of the studied organs. Conclusion and clinical importance: The distribution of glucagon- and insulin-immunopositive endocrine cells in the porcine extrahepatic biliary tract was established for the first time as a new source of these hormones. The presence of α- and ß-cells in the epithelium of extrahepatic bile ducts can be applied in treatment of diabetes, taking into account the possibility to reprogram the biliary epithelium to mentioned pancreatic endocrine cell types.

Role of dendritic cells in progression and clinical outcome of colon cancer.

PURPOSE: The dendritic cells (DCs) are key players in the initiation and regulation of immune responses including antitumor immunity. In the current study, we aimed to elucidate the role of different subtypes of DCs infiltrating the tumor stroma and invasive margin for tumor progression and survival of patients with colon cancer. METHODS: The presence of immature (CD1a- and S100 protein+) and mature (CD83- and HLA-DR+) DCs was evaluated by immunohistochemistry in tissue samples from 145 patients with colon cancer. Patients were dichotomized according to the number of DCs in the tumor stroma and invasive margin, and clinical, histological, and survival data were compared between the two groups of patients. RESULTS: The number of the mature CD83+ DCs in the tumor stroma and in the invasive margin significantly correlated with the tumor stage: the lower level of infiltration was found in patients that have advanced tumor stage. The frequency of distant metastases was higher in patients who had lower numbers of immature CD1a+ DCs in tumor stroma and of CD83+ DCs in invasive margin. Patients showing a relatively high number of S100+ DCs in the tumor stroma and HLA-DR+ DCs in the invasive margin had a longer overall survival (p < 0.05). Patients with lower CD83+ DCs infiltration in invasive margin had worse prognosis after surgical therapy compared with those with higher CD83+ DCs infiltration (p = 0.0397). CONCLUSIONS: Our results demonstrate that the infiltration of colon cancer with DCs is related with tumor progression and patient prognosis, suggesting a central role for DCs in controlling local antitumor immunity.

Functional genetic polymorphisms in interleukin-12B gene in association with systemic lupus erythematosus.

Genetic polymorphisms in cytokine genes, which influence gene expression, may have an important impact on SLE susceptibility and severity. The aim of this study was to examine the possible influence of two functional polymorphisms in cis-regulatory regions of IL12B gene in the susceptibility and clinical symptoms of SLE in Bulgarian patients. Female SLE patients (n = 141) and 124 healthy women were included in the study. Genotyping for the IL12B A/C polymorphism in 3'UTR was performed by restriction fragment length polymorphisms PCR assay and for the IL12Bpro polymorphism by allele-specific PCR. Genotype-22 of IL12Bpro polymorphism was overrepresented among women with SLE (28 vs. 17%; OR = 1.875; 95% CI: 1.037 ÷ 3.390; P = 0.037). Respectively, a higher frequency of allele-2 was found in patients than in controls (51% vs. 40%; OR = 1.566; 95% CI: 1.110 ÷ 2.210; P = 0.011). Also, we found significantly elevated frequency of genotype-22 of IL12Bpro polymorphism among SLE patients who were simultaneously carrier of genotype-AA of SNP in 3'UTR. Women with both homozygous genotypes, genotype-AA of SNP in 3'UTR and genotype-22 of IL12Bpro polymorphism, had a 2.1-fold significantly elevated risk for SLE development. The carrying of genotype-11 of IL12Bpro polymorphism was positively associated with hematological and neuropsychiatric manifestations of SLE. We have provided evidence that the IL12Bpro polymorphism was associated with SLE development among Bulgarian women. Although a strong individual effect of SNP in 3'UTR of IL12B was not detected, we found that genotype-22 of IL12Bpro was predominantly combined with genotype-AA of SNP in 3'UTR among SLE patients and this combination elevates the risk of SLE.

Angiolymphoid hyperplasia with eosinophilia--an incidental finding after surgical excision.

Angiolymphoid hyperplasia with eosinophilia (ALHE) is a rare benign vasoproliferative lesion. Although it is a benign disease, lesions are often persistent and difficult to eradicate. ALHE typically presents clinically as papules or nodules, tan, brown, pink or dull red in colour, located predominantly in the head and neck region, especially around the ears and on the forehead and scalp.All races can be affected and no gender predominance exists. The disease also has nonspecific clinical features, hence it requires in most of the cases biopsy for accurate diagnosis. We present an uncommon clinical presentation of the disease, mimicking clinically a subcutaneous lipomatous mass, which has been treated successfully with surgical excision.

Prognostic significance of CD83 positive tumor-infiltrating dendritic cells and expression of TGF-beta 1 in human gastric cancer.

BACKGROUND/AIMS: In this study we analyzed the significance of CD1a and CD83 positive tumor infiltrating dendritic cells (TIDCs) and the expression of TGF-ß1 in gastric cancer tissue, and their relationship with disease progression and prognosis of patients. METHODOLOGY: The immunohistochemical expression of CD1a, CD83 and TGF-ß1, was evaluated in 55 patients with gastric cancer and followed-up for five years. RESULTS: We found tumor infiltration with CD1a and CD83 positive DCs in all 55 cases and cytoplasmic TGF-ß1 immunoreactivity in tumor cells in 76.4% of cases. TGF-ß1 expression correlated to low CD83 positive DCs in 100% of the samples (χ2=7.66; p=0.022). Low CD83 positive DCs in tumor border (χ2=15.38; p<0.001) was also observed in 100% of tumors with TGF-ß1 expression. The number of CD1a and CD83 positive TIDCs in the tumor border was inversely correlated with positive lymph node metastases (χ2=6.64; p=0.036 and χ2=6.44; p<0.04, respectively). Patients with a low number of tumor infiltrating CD83 positive DCs had shorter survival rates (p=0.022) and patients with TGF-ß1 expression had a worse prognosis after surgical therapy (p=0.017). CONCLUSIONS: Our results suggest that tumor infiltration with DCs may be of great importance in initiating the primary anti-tumor immune response. In patients with resectable gastric cancer, the grade of TIDCs and TGF-ß1 expression could be a useful predictor of prognosis.

Expression of the xenobiotic- and reactive oxygen species-detoxifying enzymes, GST-pi, Cu/Zn-SOD, and Mn-SOD in the endocrine cells of colorectal cancer.

BACKGROUND AND AIM: The aim of the present work was to analyze the expression of antioxidant enzymes GST-pi, SOD1, and SOD2 in endocrine cells of colorectal cancers and to evaluate the significance of the presence of thus labeled endocrine cells as prognostic factor. METHODS: The expression of chromogranin A (ChGA), GST-pi, SOD1, and SOD2 was determined in endocrine cells of 128 colorectal cancers using light and electron immunohistochemistry and double immunogold labeling method. RESULTS: Endocrine cells expressing at least one of the studied antioxidant enzymes were detected in a relatively small proportion of primary colorectal cancers (22 cases, 17%; 14% GST-pi-positive, 14% SOD1-positive, and 9% SOD2-positive). The double immunogold staining and the following electron microscopy showed that GST-pi, SOD1, and SOD2 were co-localized with ChGA to the granules of most endocrine cells. The survival analyses revealed that patients with endocrine cells in primary tumor tissues expressing GST-pi had worse prognosis after the surgical therapy than those without GST-pi-positive endocrine cells (median of 22.70 vs. 49.43 months, p < 0.05, Log-rank test). CONCLUSIONS: Most of the ChGA-positive endocrine cells in colorectal cancers also expressed some or all of the three studied antioxidant enzymes, GST-pi, SOD1, and SOD2. Moreover, patients having tumors with GST-pi-positive endocrine cells have an unfavorable prognosis. We suggest that not the neuroendocrine differentiation in general, but the presence in the tumors of endocrine cells with activated antioxidant defense and probably metabolically more active might determine a more aggressive type of cancer leading to worse prognosis for patients.

Role of TGF-beta1, its receptor TGFbetaRII, and Smad proteins in the progression of colorectal cancer.

AIM: In the current study, we investigated the expression of TGF-beta1, its receptor TGFbetaRII, and the signaling proteins Smad4 and Smad7 in colorectal cancer tissue in relation to infiltration with antigen-presenting cells and some clinical and pathologic parameters of disease progression in patients with colorectal cancer (CRC). MATERIALS AND METHODS: The immunohistochemical expression of TGF-beta1, TGFbetaRII, Smad4, Smad7, HLA-DR antigen, CD1a, CD83, and CD68 was evaluated in 142 patients (50 females and 92 males) with CRC, followed-up for 6-8 years period. RESULTS: In our study, 127 (89.4%) out of 142 colorectal cancers displayed cytoplasmic TGF-beta1 immunoreactivity. Common-mediator Smad4 was detected in the tumor cytoplasm in 124 cancers (79.5%) and inhibitory Smad7 immunostaining was observed in 110 (77.4%) tumor specimens. TGFbetaRII was expressed on tumor cell membranes in 119 (76.3%) of the cancers. The increased TGF-beta1 expression in tumor cytoplasm was related to low CD68(+)- and CD83(+)-cell infiltration in tumor tissues. Patients with TGF-beta1 overexpression had worse prognosis after surgical therapy compared to those with low expression of TGF-beta1. The observed association was more pronounced for the patients in T1-T2 stage (p = 0.0015). CONCLUSIONS: The expression of TGF-beta1, its receptor TGFbetaRII, and signaling proteins Smad4 and Smad7 was observed in the majority of colorectal cancer specimens. Our results suggest that TGF-beta1 production by tumor cells may affect the tumor environment via suppression of tumor-infiltrating immune cells and probably contributes to tumor cells aggressiveness through autocrine activation of Smad signaling.