Immunoexpression of Bmi-1, CK15, Bcl-2 in different types of basal cell carcinomas.

Introduction: Basal cell carcinoma (BCC) occurs in aggressive and non-aggressive forms. The expression of immunohistochemical markers varies in different types of BCC. Aim: Immunohistochemical analysis of selected proteins in BCCs. Material and methods: The immunohistochemical method was used to examine the immunoexpression of Bmi-1, CK15 and Bcl-2 in 56 cases of BCC divided into four groups. Results: Positive Bmi-1 staining 3-4+ level (nodular type) was seen in 91.3% of samples, 4+ (infiltrative) in 92.3%, 4+ (nodular/infiltrative) - 69.2%, 3+ - 30.8%, in BSC 3+ - 42.8%, and 28.6% each for 2+ and 4+. Low grade positivity (0-1+) in CK15 staining was present in 52.1% of nodular BCC, 46.2% - nodular/infiltrative, 92.3% - infiltrative, and 100% - BSC, but levels 2-3+ in nodular BCC in 47.8%, nodular/infiltrative BCC - 53.8%, infiltrative - 7.7%. Bcl-2 positivity (3-4+) was revealed in nodular BCC in 95.6%, (1-2+) in 100% of BSC, infiltrative and infiltrative/nodular BCC, but the lowest (0-1+) in 76.9% of nodular/infiltrative BCC, 71.4% of BSC, and in 38.4% of infiltrative BCC. Conclusions: Positive Bmi-1 staining was the highest in the aggressive infiltrative subtype of BCCs, whereas the lowest in basosquamous cell carcinomas (BSC). Infiltrative BCC was characterized by a lower level of CK15 expression than nodular BCC and nodular/infiltrative BCC. Differentiation of Bcl-2 expression depended on the type of tumour; the highest level was found in nodular BCC, low grade in nodular/infiltrative and infiltrative BCCs, and BSC.

Close relationship between TAZhigh/SOX2high co-localization and metastasis in oral squamous cell carcinoma.

There is growing evidence which indicates that the development and the biological features of cancer such as the invasion, metastases and recurrence are related to the presence and behavior of the cancer stem cells (CSC). However, the regulatory mechanisms underlying CSCs-specific properties are poorly determined, the Hippo pathway has emerged as a fundamental regulator underlying CSCs stemness. Immunohistochemical method was used to examine the immunoexpression of SOX2, TAZ and α-SMA in oral squamous cells carcinomas: with metastases - OSCC M+ (n = 42), and without metastases - OSCC M- (n = 44), and 17 control cases. The immunoexpression of SOX2, TAZ and α-SMA was significantly increased in both group of OSCC in comparison to control groups. Moreover, significantly increased TAZ and α-SMA immunoexpression were found in OSCC M+ compared to OSCC M-. In OSCC M+ and OSCC M- groups there were statistically significant correlations between the immunoexpression of TAZ vs SOX2 (r = 0.56, p < 0.001; r = 0.33, p < 0.03 respectively), and TAZ vs α-SMA (r = 0.64, p < 0.001; r = 0.67, p < 0.001 respectively). Moreover, there was statistically significant association between TAZ high /SOX2 high coexistent immunoexpression and the presence of metastases (p < 0.007). Our results may suggest that SOX2 and TAZ could potentially cooperate and contribute to process of metastasis, especially in cases with TAZ high /SOX2 high expression.

Overexpression of cathepsin K and vascular endothelial growth factor in chronic venous ulcerations.

INTRODUCTION: Chronic venous disease (CVD) is a disabling condition affecting about 1% to 3% of the general population. Besides varicose veins, CVD can result also in the formation of severe skin lesions, especially venous ulcerations (VU). The exact mechanism of VU is still unknown. AIM: To evaluate immunoexpression of vascular endothelial growth factor (VEGF) and cathepsin K in healthy individuals and patients with VU. MATERIAL AND METHODS: The study included 12 patients with venous ulcers and 10 healthy individuals who served as controls; both groups were sex- and age-matched. Biopsy samples were obtained from lower leg areas and submitted to histochemical analysis. RESULTS: There was a significant difference between the study group and the control group in cathepsin K expression (1.007 ±0.3 vs. 0.22 ±0.2, respectively, p < 0.001) and VEGF expression (1.17 ±0.59 vs. 0.27 ±0.19, respectively, p < 0.001). Additionally, the microvessel density (per mm2) differed significantly between the study group and the control group (97.6 ±28.81 vs. 59.32 ±12.71, respectively, p < 0.001). We found no correlation between cathepsin K and microvessel density, and cathepsin K and VEGF in both groups, but there was a significant correlation between microvessel density and VEGF immunoexpression in the study group (r = 0.82, p = 0.002). CONCLUSIONS: Increased immunoexpression of VEGF and cathepsin K suggests that both of these proteins may play a role in VU development.

The number of CD163 positive macrophages is associatedwith more advanced skin melanomas, microvessels density and patient prognosis.

The study was aimed to evaluate the number of TAMs and to investigate whether they have association with microvessels density and patients' survival times. 46 cases of melanomas, divided into four groups according to the Breslow scale, were tested immunohistochemically with antibodies anti-CD68, CD163, iNOS to vizualized macrophages and anti-CD34 antibody to stain microvessels. The number of macrophages and the microvessels density were counted by hotspot analysis using an image analysis system. The study revealed increased numbers of CD68 and CD163 positive macrophages in successive stages of Breslow scale, but statistically significant differences were observed only between I and IV group for CD68 positive macrophages, and between I and III, IV group for CD163 positive macrophages. The mean number of the microvessels was significantly increased in group II, III, IV compared to group I. The correlative study showed significant positive correlations between the mean number of CD68 and CD163 positive macrophages and microvessels density. Moreover, the number of CD163 positive macrophages was associated inversely with patient's survival time. The results of our study may indicate that higher infiltration of macrophages, especially CD163 positive cells, is associated with more advanced melanomas, microvessels density and worse patient's prognosis.

T cells are involved in the induction of macrophage phenotypes in oral leukoplakia and squamous cell carcinoma-a preliminary report.

BACKGROUND: The prognosis of human malignancies has been shown to depend on immunological parameters, such as macrophage polarization (M1 and M2). In this study, we identify the phenotype of macrophages, and investigate an involvement of infiltrated T cells that participate in the polarization of macrophages, in oral leukoplakia (OLK), and oral squamous cell carcinoma (OSCC). METHODS: Immunohistochemical method was used to examine the number of CD68+ , CD163+ (M2), iNOS+ (M1) macrophages, and CD4+ , CD8+ , CCR4+ (Th2), CCR5+ (Th1) cells in 102 cases of OSCC: without metastases-OSCC M(-) (n = 54), and with metastases-OSCC M(+) (n = 48), 23 cases of OLK, and 18 control cases. RESULTS: The mean number of CD68+ , CD163+ , iNOS+ , CD4+ , CCR4+ , CCR5+ cells was significantly increased in OSCC M(+) group compared with OLK, OSCC M(-) and control group. We found positive correlations between the number of CD4+ T cells and CD163+ and iNOS+ macrophages as well as CCR4+ and CCR5+ cells in both OSCC groups. The mean number of CD8+ cells was significantly increased in OSCC M(-) and OLK compared with OSCC M(+) and control group. In OSCC M(+) and OSCC M(-) groups, a negative correlation between the number of CD8+ cells and CD163+ and iNOS+ macrophages was found. CONCLUSIONS: The number and co-localization of lymphocytes and macrophages in OLK and OSCC may indicate that infiltrating cells influence the early and subsequent stage of oral carcinogenesis.

Overexpression of ADAM10 in oral squamous cell carcinoma with metastases.

ADAMs (a disintegrin and metalloproteinase) are important mediators of cell signalling events, which play a role in the pathogenesis and progression of cancers. Immunohistochemical method was used to examine the immunoexpression of ADAM10 and microvessel density in 80 cases of oral squamous cell carcinoma (OSCC): without metastases - OSCC M(-) (n = 38), and with metastases - OSCC M(+) (n = 42), in 24 cases of oral leukoplakia (OLK), (15 cases with low-grade dysplasia - OLK-LG, and 9 cases with high-grade dysplasia - OLK-HG), and 19 controls. The immunoexpression of ADAM10 and the mean number of vessels were significantly increased in both groups of OSCC in comparison to both groups of OLK and controls. Moreover, the immunoexpression of ADAM10 and microvessel density were significantly increased in the OSCC M(+) group in comparison to the OSCC M(-) group. No statistically significant differences were found between immunoexpression of ADAM10 and microvessels density in the OLK-LG, OLK-HG, and control cases. In conclusion, the present study revealed overexpression of ADAM10 in OSCCs, especially in OSCC with metastasis. These findings suggest that ADAM10 could potentially contribute to metastases of oral cancer. Although, our findings suggest that ADAM10 may be involved in angiogenesis of OSCC, further studies are required to determine the role of ADAM10 in this process.

Association of infiltrating cells with microvessel density in oral squamous cell carcinoma.

Several lines of evidence indicate that immune cells in the tumor microenvironment play an important role in regulating tumor progression. An immunohistochemical method was used to examine the abundance of natural killer (NK) cells, mucosal dendritic cells (DCs), macrophages, mast cells, and microvessel density in 78 cases of oral squamous cell carcinoma (OSCC): with better prognosis - OSCCBP (n = 37), and with poorer prognosis - OSCCPP (n = 41), and 18 controls. The mean numbers of macrophages and microvessels were significantly higher in the OSCCPP group in comparison to both OSCCBP and control groups. The mean number of NK cells, mast cells and DCs was lower in the OSCCPP group in comparison to the OSCCBP group, but there were no statistically significant differences between mean numbers of NK cells in tested groups. Statistically significant correlations between the number of DCs and NK cells and mast cells, as well as between microvessel density and numbers of macrophages, DCs and mast cells were revealed in both OSCCPP and OSCCBP groups. In conclusion, our findings revealed an association between the number of infiltrating cells and oral cancer prognosis. Moreover, our results suggest that the infiltrating cells (macrophages, Langerhans and mast cells) may be involved in the process of angiogenesis.

Gene/protein expression of CAPN1/2-CAST system members is associated with ERK1/2 kinases activity as well as progression and clinical outcome in human laryngeal cancer.

Recent evidence indicates the involvement of calpains (CAPNs), a family of cysteine proteases, in cancer development and progression, as well as the insufficient response to cancer therapies. The contribution of CAPNs and regulatory calpastatin (CAST) and ERK1/2 kinases to aggressiveness, disease course, and outcome in laryngeal cancer remains elusive. This study was aimed to evaluate the CAPN1/2-CAST-ERK1/2 enzyme system mRNA/protein level and to investigate whether they can promote the dynamic of tumor growth and prognosis. The mRNA expression of marker genes was determined in 106 laryngeal cancer (SCLC) cases and 73 non-cancerous adjacent mucosa (NCLM) controls using quantitative real-time PCR. The level of corresponding proteins was analyzed by Western Blot. SLUG expression, as indicator of pathological advancement was determined using IHC staining. Significant increases of CAPN1/2-CAST-ERK1/2 levels of mRNA/protein were noted in SCLC compared to NCLM (p < 0.05). As a result, a higher level of CAPN1 and ERK1 genes was related to larger tumor size, more aggressive and deeper growth according to TFG scale and SLUG level (p < 0.05). There were also relationships of CAPN1/2 and ERK1 with incidences of local/nodal recurrences (p < 0.05). An inverse association for CAPN1/2, CAST, and ERK1/2 transcripts was determined with regard to overall survival (p < 0.05). In addition, a higher CAPN1 and phospho-ERK1 protein level was related to higher grade and stage (p < 0.05) and was found to promote worse prognosis. This is the first study to show that activity of CAPN1/2- CAST-ERK1/2 axis may be an indicator of tumor phenotype and unfavorable outcome in SCLC.

Metallothionein 2A core promoter region genetic polymorphism and its impact on the risk, tumor behavior, and recurrences of sinonasal inverted papilloma (Schneiderian papilloma).

Inverted papillomas are a unique group of locally aggressive benign epithelial neoplasms in the nasal cavity and paranasal sinuses arising from the Schneiderian mucosa. Metallothioneins are sulfhydryl-rich heavy metal-binding proteins required for metal toxicity protection and regulation of biological mechanisms including proliferation and invasion. The goal of this study was to identify three SNPs at loci -5 A/G (rs28366003) and -209 A/G (rs1610216) in the core promoter region and at locus +838 C/G (rs10636) in 3'UTR region of the MT2A gene with IP risk and with tumor invasiveness according to Krouse staging. Genotyping was performed using the PCR restriction fragment length polymorphism technique in 130 genetically unrelated IP individuals, and 418 randomly selected healthy volunteers. The presence of the rs28366003 SNP was significantly related to the risk of IP within the present population-based case-control study. Compared to homozygous common allele carriers, heterozygosity and homozygosity for the G variant had a significantly increased risk of IP (adjusted odds ratio [OR] = 7.71, 95% confidence interval [CI]: 4.01-14.91, p(dominant) < 0.001). Moreover, risk allele carriers demonstrated higher Krouse stage (pT1 vs. pT2-4) (OR = 19.32; 95% CI, 2.30-173.53; p < 0.0001), diffuse tumor growth (OR = 4.58; 95% CI, 1.70-12.11; p = 0.0008), bone destruction (OR = 4.13; 95% CI, 1.50-11.60; p = 0.003), and higher incidence of tumor recurrences (OR = 5.11; 95% CI, 1.68-15.20; p = 0.001). The findings suggest that MT2A gene variation rs28366003 may be implicated in the etiology of sinonasal inverted papilloma in a Polish population.

Augmented immunoexpression of survivin correlates with parameters of aggressiveness in prostate cancer.

The clinical significance of the immunoexpression of survivin in prostate cancer and its correlation with the biological aggressiveness of prostate cancer remains unclear. Therefore, the present study was undertaken to compare the immunoexpression of survivin in prostate cancer (PCa) and benign prostatic hyperplasia (BPH) as well as to determine whether this immunoexpression could correlate with Gleason score, proliferation activity and prostate specific antigen (PSA) levels. The prostate needle biopsies from 28 patients with elevated serum PSA levels were studied. As a control, 12 needle biopsies of prostate diagnosed as BPH were used. The immunoexpression of survivin was evaluated semiquantitatively, whereas the Ki-67 index was assessed quantitatively. The immunoexpression of survivin and Ki-67 in epithelial cells in the prostate cancer group was significantly increased as compared to BPH cases. In the prostate cancer group there were positive significant correlations between the immunoexpression of survivin and Gleason score as well as Ki­67 antigen. The correlation between the immunoexpression of survivin and PSA levels was also positive, but it did not reach statistical significance. In conclusion, we can confirm that in prostate cancer the immunoexpression of survivin is augmented as compared to BPH and positively correlated with parameters of tumor aggressiveness.

Mediators of mast cells in bullous pemphigoid and dermatitis herpetiformis.

Bullous pemphigoid (BP) and dermatitis herpetiformis (DH) are skin diseases associated with inflammation. However, few findings exist concerning the role of mast cells in autoimmune blistering disease. Skin biopsies were taken from 27 BP and 14 DH patients, as well as 20 healthy individuals. Immunohistochemistry was used to identify the localization and mast cell expression of TNFα and MMP9 in skin lesions and perilesional skin. The serum concentrations of TNFα, MMP9, chymase, tryptase, PAF, and IL-4 were measured by immunoassay. TNFα and MMP9 expression in the epidermis and in inflammatory influxed cells in the dermis was detected in skin biopsies from patients. Although these mediators were found to be expressed in the perilesional skin of all patients, the level was much lower than that in lesional skin. Increased serum PAF levels were observed in BP patients. Mast cells may play an essential role in activating inflammation, which ultimately contributes to the tissue damage observed in BP and DH. Our findings suggest that differences in the pattern of cytokine expression directly contribute to variations in cellular infiltration in DH and BP.

An assessment of the (anti)androgenic properties of hexachloronaphthalene (HxCN) in male rats.

The persistent organic pollutants (POPs) defined by the Stockholm Convention include polychlorinated naphthalenes (PCNs); of these, the most toxic, persistent, abundant, dioxin-like congeners found in human tissues are the hexachloronaphthalenes (HxCNs). Recent research also indicates that PCNs may disrupt hormonal homeostasis. The aim of this study was to evaluate the (anti)androgenic action of HxCN. Immature, castrated male Wistar rats were exposed per os to HxCN in corn oil at daily doses ranging from 0.3 to 3.0 mg kg-1 for 10 days. According to the OECD 441 protocol (Hershberger Bioassay), the anti-androgenic assay groups were co-exposed with testosterone propionate (TP), while the androgenic groups were not. TP was used as the reference androgen (subcutaneous daily doses of 0.4 mg kg-1), and flutamide (FLU) as the reference antiandrogen (per os daily doses of 3.0 mg kg-1). Five assessory sex tissues (ASTs) were weighed: ventral prostate, seminal vesicles, levator ani-bulbocavernosus muscle (LABC), Cowper's glands and glans penis. HxCN + TP significantly decreased the weight of the ventral prostate and seminal vesicle indicating an anti-androgenic action via 5α-reductase inhibition. These weight changes were also accompanied by abnormalities in cell morphology and hormonal disturbances: lowered levels of the testosterone and thyroid hormones thyroxine and triiodothyronine. Disturbances were also noted in the lipid profile, viz. total cholesterol, triglycerides and high-density lipoprotein and non-HDL fraction content. However, the direction of these changes differed depending on the size of the HxCN dose. No dose-effect relationship was noted for most of the obtained results; as such, exposure to even small HxCN doses run the risk of anti-androgenic effects in the general population, especially when encountered in combination with other POPs and endocrine-disrupting chemicals in the environment.

IL-17 expression in dermatitis herpetiformis and bullous pemphigoid.

Dermatitis herpetiformis (DH) and bullous pemphigoid (BP) are skin diseases associated with eosinophilic and neutrophilic infiltrations. Although cytokines are critical for the inflammatory process, there are single findings concerning concentration of IL-17 in bullous diseases. The goal of this study was to assess IL-17 expression in DH and BP patients. Skin biopsies were taken from 10 DH, 14 BP patients and from 10 healthy subjects. The localization and expression of IL-17 was studied by immunohistochemistry and the serum concentration was measured by immunoassays. Expression of IL-17 in the epidermis and in influxed cells in dermis was detected in skin biopsies. Expression of IL-17 was statistically higher in epidermis and infiltration cells in specimens from BP than from DH patients. Examined interleukin expression was detected in perilesional skin of all patients but it was much lower than in lesional skin. The expression of IL-17 was not observed in biopsies from healthy people. Serum level of IL-17 was statistically higher in BP and DH groups as compared to control group. Our results provide the evidence that IL-17 may play an essential role in activating and recruiting eosinophils and neutrophils, which ultimately contribute to the tissue damage in DH and BP.

Augmented mast cell infiltration and microvessel density in prostate cancer.

AIM OF THE STUDY: Recent investigations have taken into account the role of mast cells in prostate cancer formation, analyzing their dual functions (as tumour growth promoters and tumour growth inhibitors). The aim of our study was to compare mast cell infiltration and microvessel density in prostate cancer and in benign prostate hyperplasia. We also attempted to find possible relationships among mast cell infiltration and microvessel density, Gleason score, as well as serum levels of prostate-specific antigen (PSA). MATERIAL AND METHODS: The investigation was confined to evaluations of material from prostate needle biopsies, carried out in 26 patients with prostate cancer, and of 14 specimens diagnosed as benign hyperplasia. The numbers of tryptase positive mast cells and CD34 positive vessels were determined using a computer image analysis system. In the patients with prostate cancer, both mast cell infiltrates and microvessel density were significantly increased, as compared to the control patients. RESULTS: Significant positive correlations were identified between the mean numbers of mast cells and microvessel densities, both in the prostate cancer group and in the control group. Moreover, significant positive correlations were observed between Gleason score on one hand and the number of mast cells and microvessel density on the other. The correlations between PSA serum levels and both mast cell infiltration and microvessel density were positive, but not in a statistically significant way. CONCLUSIONS: The reported investigations may support the assumption of mast cell promoter function in prostate cancer development, whereas no evidence was found for their opposite.

Expression of arachidonate metabolism enzymes and receptors in nasal polyps of aspirin-hypersensitive asthmatics.

BACKGROUND: The pathogenesis of rhinosinusitis in aspirin-exacerbated airway disease is closely linked to the disequilibrium in arachidonic acid metabolism. Although considerable amounts of data concerning impaired eicosanoid production are available, the precise mechanism and pathogenesis of the disease are still unknown. The aim of the present study was to assess the expression of enzymes belonging to the arachidonic acid cascade and receptors for arachidonate derivative metabolites in nasal polyps from aspirin- hypersensitive (AH) and aspirin-tolerant (AT) patients with rhinosinusitis. METHODS: Cells expressing cysteinyl leukotriene (CysLT) receptors (CysLT(1) and CysLT(2)), arachidonate 5-lipoxygenase, leukotriene B(4) receptor type 1, E-prostanoid receptors (EP(2) and EP(4)), cyclooxygenase (COX)-1 and COX-2 were detected by immunocytochemistry in nasal polyps obtained from 10 AH patients and 18 AT patients. RESULTS: There was a significantly higher density of cells expressing CysLT(1) and CysLT(2) receptors in nasal polyps from AH patients than from AT patients (p < 0.001). In contrast, the density of cells expressing EP(2) receptor and COX-2 was significantly lower in AH patients than in AT patients (p < 0.02). The number of COX-2-positive epithelial cells was significantly reduced in AH polyps (p < 0.04). CONCLUSIONS: The elevated number of nasal polyp cells expressing CysLT receptors and lack of cells expressing EP(2) receptor and COX-2 may be related to a more severe course of hyperplastic rhinosinusitis in aspirin hypersensitivity.

The immunoexpression of Shh, Smo and Gli2 in Helicobacter pylori positive and negative gastric biopsies.

The Hedgehog signaling pathway plays a principal role during embryonic development, tissue regeneration and carcinogenesis in various adult tissues. Although hedgehog signaling is important in gastric carcinogenesis, its role in Helicobacter pylori-associated gastritis is unclear. The aim of our study was to examine Sonic Hedgehog (Shh) signaling pathway in response to H. pylori infection. Thirty-one formalin-fixed, paraffin-embedded tissue specimens of chronic gastritis were retrieved from archival material. The immunoexpression of Shh, Smoothened (Smo) and Glioblastoma transcription factor 2 (Gli2) were detected using the immunohistochemical method. Sonic Hedgehog protein was expressed in H. pylori-positive and H. pylori-negative groups of patients. The immunoexpression of Shh, Smo and Gli2 proteins was lower in H. pylori-positive group compared to H. pylori-negative group, however only the differences in Shh and Smo immunoexpression were statistically significant. The immunoexpression of Shh was significantly correlated with the immunoexpression of Smo in both tested groups (p < 0.001, p < 0.02, respectively). No statistically significant correlation was found between Shh and Gli2 among H. pylori-positive and H. pylori-negative groups. The above findings support the hypothesis of the involvement of Shh signaling pathway in H. Pylori-associated gastritis.

Expression of selected neuropeptides in pathogenesis of bullous pemphigoid and dermatitis herpetiformis.

Bullous pemphigoid (BP) and dermatitis herpetiformis (DH) are chronic subepidermal bullous diseases, which progress together with an itch and an inflammatory reaction. These symptoms may be the cause of a phenomenon described in the literature as a neurogenic skin inflammation. Neuropeptides are one of the mediators which take part in this process. The aim of our study was to indicate the expression of selected neuropeptides - CRF (corticotropin releasing factor), CGRP (calcitonin gene-related peptide), NKB (neurokinin B), SP (substance P) and the receptor for endothelin B (ETRB) - in the skin of patients suffering from BP or DH. A significantly increased expression of CRF was found in the specimen collected from the skin lesions of patients with BP and DH as well as a significantly increased expression of receptor for endothelin B in the patients with DH by the immunohistochemical method. The results obtained give evidence of a possible participation of CRF and receptor for endothelin B in the pathogenesis of the itch in the dermatitis herpetiformis as well as CRF in bullous pemphigoid.

Snail Overexpression Alters the microRNA Content of Extracellular Vesicles Released from HT29 Colorectal Cancer Cells and Activates Pro-Inflammatory State In Vivo.

During metastasis, cancer cells undergo phenotype changes in the epithelial-mesenchymal transition (EMT) process. Extracellular vesicles (EVs) released by cancer cells are the mediators of intercellular communication and play a role in metastatic process. Knowledge of factors that influence the modifications of the pre-metastatic niche for the migrating carcinoma cells is important for prevention of metastasis. We focus here on how cancer progression is affected by EVs released from either epithelial-like HT29-cells or from cells that are in early EMT stage triggered by Snail transcription factor (HT29-Snail). We found that EVs released from HT29-Snail, as compared to HT29-pcDNA cells, have a different microRNA profile. We observed the presence of interstitial pneumonias in the lungs of mice injected with HT29-Snail cells and the percent of mice with lung inflammation was higher after injection of HT29-Snail-EVs. Incorporation of EVs released from HT29-pcDNA, but not released from HT29-Snail, leads to the increased secretion of IL-8 from macrophages. We conclude that Snail modifications of CRC cells towards more invasive phenotype also alter the microRNA cargo of released EVs. The content of cell-released EVs may serve as a biomarker that denotes the stage of CRC and EVs-specific microRNAs may be a target to prevent cancer progression.

In situ hybridization for human papillomavirus.

There is increasing molecular and epidemiologic evidence that human papillomavirus (HPV) is associated with distinct subset of head and neck lesions. The aim of the present study was to evaluate a possible relationship between the presence of HPV DNA and morphological signs of HPV infections in twenty-six formalin-fixed, paraffin-embedded cases of laryngeal squamous cell carcinomas and seventeen cases of sinonasal inverted papillomas, retrieved from archival material. Ten out of twenty-six cases of cancers and eleven out of seventeen sinonasal inverted papillomas showed morphological signs of HPV infections. All cases have been analyzed using in situ hybridization. The expression of HPV DNA was noted in both examined groups of squamous cell carcinomas and in group of sinonasal inverted papillomas when the morphological signs of HPV infections were present. No significant differences in the frequency of HPV DNA expression were noted in both examined groups of cancers and inverted papillomas (with and without morphological signs of HPV infection). In conclusion, our study confirms the role of HPVs in pathogenesis of inverted papillomas and laryngeal squamous cell carcinomas. However, there is no relevant relationship between the presence of HPV DNA and morphological signs of HPV infections in studied cases.