Reduced FHIT expression is associated with tumor progression in sporadic colon adenocarcinoma.

PURPOSE: Tumor supressor gene FHIT was identified at chromosome 3p14.2 spanning the FRA3B fragile site and is very often inactivated in different types of cancer. The aim of this study was to examine the frequency of FHIT gene LOH as well as FHIT mRNA and protein expression in sporadic colon adenocarcinoma. METHODS: The results of LOH, real-time qRT-PCR and imunohistochemical analyses were correlated with clinico-pathological characteristics of patients and their tumors in order to evaluate the role of FHIT gene/protein in sporadic colon adenocarcinoma tumorigenesis. RESULTS: One hundred and thirty one (96.3%) samples were informative for both markers and 33/131 (25.2%) demonstrated LOH. Expression of FHIT mRNA was significantly decreased in colon tumors relative to that in corresponding normal tissue (p = 7.2×10(-6)). Most of the samples (54.0%) were negative for FHIT protein, 26.4% adenocarcinomas showed a weak to moderate immunostaining and 19.6% adenocarcinomas showed strong FHIT immunostaining. No correlation was found between FHIT gene LOH status, mRNA expression or FHIT protein immunostaining and clinico-pathological characteristics. Expression of FHIT mRNA was significantly decreased in FHIT LOH positive tumors (p = 0.027). Patients with LOH negative tumors or FHIT protein positive tumors had longer survival but this findings were not statistically significant. CONCLUSIONS: Our overall results suggest that reduced expression of FHIT gene may be associated with the progression of these malignant tumors.

An unusual mole: an adult case of Dabska tumour.

In 1969 Dabska and her colleagues described for the first time this rare malignant tumour, also later known as a malignant endovascular papillary angioendothelioma of childhood. Overall, depending amongst other factors on its location, it is thought to have a generally favourable prognosis and a wide local excision seems to be the treatment of choice. We here present a very rare and unusual case of a 63 year old woman with a 20 year history of slow-growing right buccal dermatological lesion which resembled a common mole. The histopathological diagnosis of Dabska Tumour was made following the hematoxylin and eosin (H&E) biopsy. The analysis revealed multiple delicate interconnecting vascular channels with papillary plugs, some of which containing hyalinized core, projecting into the lumen lined by atypical plumped endothelial cells.

[Trends in stage and age characteristics of the female breast cancer treated in 11-year period at University Hospital "Dubrava"]. / Analiza dobne distribucije i stupnja lokalne prosirenosti raka dojke kod bolesnica operiranih u 11-godisnjem razdoblju u Klinickoj bolnici "Dubrava".

In a retrospective study we analyzed and compared the factors of local spread of breast cancer, namely tumor size and status of regional lymph nodes and patient age in the period from 1997 to 2007. The study includes only those patients in whom the diagnosis was made in an open surgical biopsy and it was possible to determine the local extent of disease at the time of diagnosis (pT and pN stage), a total of 1202 patients. We also compared the findings in patients younger and older than 50 years. The average tumor size at diagnosis was 2.2 cm and ranged between 1.7 cm (2005) and 2.4 cm (1997, 1999). The percentage of metastases in regional lymph nodes varied between 37% (2001) and 50% (2000 and 2006). The average of 6% of all cases of breast cancer was diagnosed in non-invasive (in situ) stage. The mean patient age was 56.8 years and for the observed age ranged between 54.3 (1999) and 59.1 (2004) years. There was no statistically significant difference in relation to disease stage or patient age at diagnosis observed throughout the entire period of time. There was also no statistical difference in stage of disease at the time of diagnosis in patients younger and older than 50 years.

Synchronous bilateral breast carcinoma with two different morphology subtypes: a case report.

We report a case of synchronous bilateral breast cancer with ductal and medullary carcinoma. A 60-year-old woman presented with lesion in both breasts which were mammographically found two years ago. Ultrasonography proved two suspected masses in breasts. Fine needle cytology was performed and confirmed bilateral carcinoma but with different cytological findings. The cytological feature of the left breast suggested ductal carcinoma and of the right breast raised possibility of a medullary carcinoma. Patient underwent bilateral quadrectomy with evacuation of axillary lymph nodes. Histological examination showed bilateral carcinoma with two different histological features: ductal in the left and medullary carcinoma in the right breast.

Intraoperative imprint cytological assessment of the subareolar tissue of the nipple areola complex (NAC).

One of the criteria of selection for skin sparing mastectomy (SSM) with nipple areola complex (NAC) preservation is to exclude the neoplastic involvement of subareolar tissue (NAC base) in order to minimize the possibility of local recurrence. The most common way to assess the possible neoplastic involvement is intraoperative frozen section of the NAC base tissue. Because of its limitations, particularly the false negative results due to unsampling, we tried to use intraoperative imprint cytology for more thorough intraoperative assessment. The aim was to compare intraoperative imprint findings with the definitive histology of the NAC base, to evaluate diagnostic accuracy of this method and possibility to substitute frozen section for intraoperative assessment of NAC base. A prospective clinical study was conducted of 208 consecutive female patients who underwent open biopsy because of carcinoma. Intraoperative imprints were taken from the excised subareolar tissue which was then routinely processed for definitive histology. Imprint findings designated positive, negative, suspicious or atypia, were compared with definitive histological findings. Our results with 7.5% false negative rate, 9.8% false positive rate, sensitivity of 50% and specificity of 87.58% argue that imprint cytology might not be sufficient as an exclusive method for the intraoperative assessment of the NAC base though it should be used routinely in conjunction with frozen section examination.

Influence of interleukin-8 and interleukin-10 on sporadic colon cancer development and progression.

Cytokines produced in the tumour microenvironment have an important role in cancer pathogenesis. Altered cytokine expression may result in increased susceptibility to and/or poor prognosis in certain cancers. Therefore, the aim of this study was to investigate the influence of interleukin (IL)-8 and IL-10 on sporadic colon cancer development and progression. In our study, a statistically significant increase in IL-8 messenger RNA (mRNA) expression and decrease in IL-10 mRNA expression in tumour tissue compared with normal mucous tissue was observed (P = 0.003; P = 1.3 x 10(-9)). No association was found between IL-8 -251 A/T genotypes and IL-8 mRNA expression in tumour and corresponding normal mucous tissue, as well as susceptibility to sporadic colon cancer. Positive immunohistochemical IL-8 staining was more frequent in moderately and poorly differentiated tumours compared with well-differentiated tumours (P = 0.024). Finally, IL-8 significantly stimulated invasion of HT-29 cells in vitro (P = 0.000172). Significant association of IL-10 -1082 A/G, -819 T/C and -592 A/C genotypes and IL-10 mRNA expression in tumour tissue was observed (P = 0.022; P = 0.013; P = 0.02). Significant association of -819 T/C and -592 A/C genotypes and IL-10 mRNA expression in corresponding normal mucous tissue was observed (P = 0.01; P = 0.04) as well. IL-10 single-nucleotide polymorphism (SNP) promoter genotypes associated with low IL-10 mRNA expression (-819 TT; -592 AA) were also associated with increased risk of sporadic colon cancer compared with high-expression genotypes [odds ratio, 5.53; 95% confidence interval (CI), 1.53-20.1; odds ratio, 4.07; 95% CI, 1.28-12.96]. Positive IL-10 immunohistochemical reaction was more frequent in well-differentiated and moderately differentiated tumours compared with poorly differentiated tumours (P = 0.036). In Dukes' C tumours, positive IL-10 immunohistochemical reaction was less frequent compared with Dukes' A and B tumours (P = 0.023). Taken together, our results point to possible tumour promoting role of IL-8 and potential protective role of IL-10 in sporadic colon cancer.

[Basal cell adenoma of the nasal septum: a case report]. / Adenom bazalnih stanica nosnog septuma prikaz bolesnika.

This article reports a case of an intranasal basal cell adenoma, which is an extremely rare tumor and comprises only 1 to 3% of salivary gland neoplasms. The tumor was removed by intranasal excision with the small alar rhinotomy to facilitate the access. Immunohistochemically the diagnosis of basal cell adenoma was confirmed. There was no recurrence after 24-month follow up. This location of basal cell adenoma has not yet been described in the recent literature.

Molecular genetic alterations of FHIT and p53 genes in benign and malignant thyroid gland lesions.

Several oncogenes and tumor-suppressor genes are involved either as early or late event in thyroid gland carcinogenesis. Human FHIT (fragile histidine triad) gene is highly conserved gene whose loss of function may be important in the development and/or progression of various types of cancer. We undertook this study to analyze FHIT and p53 gene status in different benignant and malignant thyroid tumors. Status of these genes as well as intensity of apoptosis was analyzed in tumor tissues by molecular genetic methods, immunohistochemistry, and FACS-scan analysis. The majority of the malignant thyroid cancers displayed aberrant expression of FHIT gene, concominant with p53 gene inactivation. This is followed by low rate of apoptosis, which may be important in the development and/or progression of thyroid cancer. We found higher incidence of p53 mutation and aberrant processing of FHIT mRNA in malignant tumors (papillary, follicular, medullary and anaplastic carcinomas) and in those tumors with distant metastasis. The growth of p53(-)/FHIT(-) follicular carcinoma of human origin was much faster in nude mice than p53(+)/FHIT(+) follicular carcinoma, and mice had shorter survival rate. Our results show a correlation between aberrant FHIT and p53 expression, low rate of apoptosis, and malignancy. Concomitant aberration of FHIT gene and p53 could be responsible for development of highly malignant types of thyroid cancer and may be considered as a prognostic marker for these tumors.

Prognostic factors of gastrointestinal stromal tumors.

Gastrointestinal stromal tumors (GIST) are rare neoplasms of unknown etiology and pathogenesis. Their clinical behavior is very unpredictable and a reliable prognostic factor is lacking. The aim of this study was to analyze some prognostic factors and estimate which one is the most reliable. Thirty-eight biopsy specimens of GIST were immunolabeled for PCNA, CD34, vimentin, NSE and actin. The greatest diameter, histological grading, mitotic count, DNA-index and S-phase were estimated for each case. All patients were followed-up for at least 24 months or to death. The data were analysed by univariate and multivariate statistical analysis using a computer program. The results showed that greatest diameter, tumor grade, mitotic count and PCNA-index are prognostic factors in univariate analysis. In multivariate analysis only the greatest diameter is a useful prognostic factor for planning further therapy.

Primary pulmonary botryomycosis: case report.

A case is presented of pulmonary botryomycosis in a 61-year-old man with a massive right-side pulmonary infiltrate which looked like a tumor (on X-ray). Microscopic examination of a transbronchial biopsy specimen revealed chronic suppurative inflammation, which did not regress despite intensive antibiotic therapy for a period of two months. Histological analysis of specimens taken during surgery for hemoptysis revealed pulmonary botryomycosis. The disease was diagnosed on the basis of characteristic eosinophilic granules in which the bacteria are surrounded by protein material (Splendore-Hoeppli phenomenon). Pulmonary actinomycosis was excluded. The case demonstrates that pulmonary botryomycosis can have the appearance of a mass which resembles pulmonary carcinoma on X-ray, and may also be mistaken for pulmonary actinomycosis. For this reason, pulmonary botryomycosis, although rare, should be excluded during differential diagnosis of hemoptysis or pulmonary infiltrates.

[Bronchiolitis obliterans with organizing pneumonia in a patient treated with amiodarone]. / Bronchiolitis obliterans s organiziranom pneumonijom u bolesnika lijecenog amiodaronom.

Bronchiolitis obliterans organizing pneumonia (BOOP) is a well-defined clinicopathological entity. The aetiology of BOOP is generally unknown, although it has been associated with specific diseases or various pharmaceutical drugs. The amiodarone is one of them. We report a patient with BOOP secondary to amiodarone therapy, who presented with cough, fever and sputum production, dyspnoea and night sweats lasting for two months. A chest radiograph showed bilateral patchy and interstitial infiltrates. Lymphocyte phenotyping of bronchoalveolar lavage fluid showed decreased ratio of CD4+:CD8+ lymphocytes. Transbronchial lung biopsy established the diagnosis of BOOP. After stopping amiodarone therapy, symptoms disappeared and the chest radiograph remained normal within two months.

[Cytodiagnosis in irradiated breast cells--its value and pitfalls]. / Citodijagnostika ozracenih stanica dojke--vrijednost i moguce pogreske.

Morphologic changes in glandular epithelium of the breast after quadrantectomy of the breast carcinoma and radiotherapy are described. The aim of the study was to establish the possibilities of cytologic assessment of morphological changes in palpable lesions in the residual breast tissue. Fine needle aspirates of 50 patients after surgery and radiotherapy of the breast carcinoma were analyzed. Biopsy and pathohistologic verification were performed in 12 patients. Carcinoma was cytologically found and confirmed pathohistologically in two cases. Recurrence was pathohistologically confirmed in 3/10 (33.3%) cytologically suspect aspirates, while in the remaining 7/10 (66.7%) suspect findings benign changes were established (2 mild ductal proliferations, 2 florid ductal proliferations, 3 cases of adenosis). The patients were followed-up clinically and cytomorphologically. No relapse was noted in the follow up period. Although its role is limited, fine needle aspiration cytology is useful in the assessment and follow-up of palpable changes after irradiation. A cytologist should be informed on whether and when a patient underwent irradiation. The finding of the changed cell after the period without changes points to the relapse of the disease.

Interleukin-6-174 G/C polymorphism is not associated with IL-6 expression and susceptibility to sporadic colon cancer.

Interleukin-6 (IL-6) has been implicated in tumorigenesis; however, its role is still far from being clearly defined. Regulation of IL-6 expression is highly complex, and additional complexity is introduced by single-nucleotide polymorphisms in the IL-6 gene. These single-nucleotide polymorphisms might influence mRNA transcription, which might in turn result in increased susceptibility to certain tumors. The aim of this study was to analyze IL-6 mRNA and protein expressions in sporadic colon cancer. Influence of IL-6-174 G/C polymorphism on IL-6 mRNA expression and sporadic colon cancer susceptibility was evaluated as well. The frequency of IL-6-174 G/C was analyzed by polymerase chain reaction-restriction fragment length polymorphism analysis. IL-6 mRNA and protein expressions were analyzed by real-time reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry. No statistically significant difference in IL-6 mRNA expression in tumor tissue compared with the corresponding normal tissue was observed (p = 0.116). No correlation was found between IL-6 mRNA and protein expressions and clinicopathological features of sporadic colon tumors. There was no association of IL-6-174 G/C genotypes with IL-6 mRNA expression in colon tumors and corresponding normal mucous tissue (p = 0.355; p = 0.152). Finally, there was no association of IL-6-174 G/C with susceptibility to sporadic colon cancer.

Altered iron metabolism, inflammation, transferrin receptors, and ferritin expression in non-small-cell lung cancer.

The involvement of iron and inflammation parameters on overall survival in non-small-cell lung cancer (NSCLC) patients was studied. Furthermore, transferrin receptors 1 (TfR1) and ferritin expression in tumor tissue, tumor stroma, and normal lung tissue were analyzed. Iron metabolism and inflammation parameters were determined by automated laboratory measurements at the time of diagnosis. TfR1 and ferritin expression were determined by immuno-histochemical methods. About 50% of patients survived 12 months only. At the time of diagnosis more than half of the patients had anemia and significantly elevated serum ferritin. Iron content of serum ferritin (ICF) was below the reference values in 90% of patients. Furthermore, ICF showed positive correlation with iron metabolic parameters and survival but negative correlation with serum ferritin and ESR. The expression of TfR1 and ferritin in tumor cells was observed in 88% or 62% of patients, respectively. Tumor stroma was TfR1 negative and sporadically ferritin positive. Tumor tissue ferritin expression showed negative correlation with serum iron and hematokrit (Ht), and positive correlation with ferritin, erythrocyte sedimentation rate (ESR), alpha-1 globulin, and alpha-2 globulin. Positive correlation was found between TfR1 expression in tumor tissue and alpha-globulin. The correlation between TfR1/ferritin expression in tumor tissue and ICF or survival was not observed. Therefore, we conclude that elevated serum ferritin in sera of NSCLC patients is the result of inflammation and oxidative stress rather than body iron overload. Higher expression of ferritin in tumor tissue may be the consequence of iron deficiency or local toxicity induced by environmental factors.

Extranotochordal extralaryngeal chordoma: a case report.

Chordomas are rare, malignant, slowly growing neoplasms which develop from vestigial remnants of the fetal notochord. Most chordomas arise in the sacrococcygeal and spheno-occipital region. Extranotochordal chordomas are extremely unusual. A case of extranotochordal chordoma with extralaryngeal localization is described. A 73-year-old male presented with swallowing difficulties and hoarseness. Contrast-enhanced magnetic resonance imaging of the neck revealed a well-encapsulated tumor mass that was well enhanced and located in the left retrolaryngeal space at the level of C4, dislocating the larynx to the right. Left radical neck dissection and tumor extirpation were performed. The tumor had not invaded cervical vertebra and the surrounding soft tissue but superficial erosions of the ossificated thyroid and cricoid cartilage were found. High-power pathologic examination and immunohistochemistry defined the lesion as a dedifferentiated type of chordoma. The patient received adjuvant radiotherapy. Four years after the surgery, the patient has been free from tumor recurrence.

TNF alpha promoter polymorphisms analysis in benign and malignant breast lesions.

Polymorphisms in genes involved in the complex mechanisms of carcinogenesis may affect the susceptibility to cancer. The multifunctional cytokine tumor necrosis factor alpha (TNF alpha) has an important role in the pathogenesis of inflammatory, autoimmune and malignant diseases. It has a large spectrum of activities, including both antitumorigenic and protumorigenic. In recent years, several TNF alpha promoter polymorphisms have been identified and related to the expression level of cytokine and to the susceptibility to solid tumors. The aim of our study was to investigate the frequency of three TNF alpha promoter polymorphisms (-1031, -308 and -238) in benign (fibrocystic changes) and malignant (invasive carcinoma) breast lesions. Using "real-time" PCR SNP analysis these polymorphisms were determined in 76 patients with benign and 158 patients with malignant breast lesions. The high expression genotypes at any of the three SNP polymorphisms were more frequent in invasive breast carcinoma (in 81 of 158 examined, 51.3%) than in fibrocystic changes (in 33 of 76 examined, 43.4%). The combined frequency of high production genotypes (-1031 T/C and C/C, -308 G/A and A/A and -238 G/A and A/A) was higher in patients with invasive breast carcinoma than in those with fibrocystic changes. However, these results were not statistically significant. Further studies on a larger group of patients are needed to evaluate the significance of potential differences in TNF alpha genotypes in different breast lesions.

Expression of c-myc, erbB-2, p53 and nm23-H1 gene product in benign and malignant breast lesions: coexpression and correlation with clinicopathologic parameters.

The aims of this study were to assess the expression of protein products of c-myc, erbB-2, p53 and nm23-H1 gene in benign and malignant breast lesions, to estimate their possible coexpression and to correlate the results of immunohistochemical analysis with various clinicopathologic parameters. The method used was the immunohistochemical detection of the corresponding protein. Expression of c-myc protein was high in both malignant and benign lesions (95% and 100%). Expression of erbB-2 and mutated p53 proteins in malignant lesions was 27% and 34%. These proteins were present in benign lesions as well: 7.8% of benign lesions were positive for erbB-2 protein and 19.6% for p53 protein. The expression of nm23-H1 protein was similar in benign and malignant lesions: 47% and 54%. The coexpression of nm23-H1 and mutated p53 protein was found in 14 carcinomas (16.5%). We found a tendency of negative correlation between the expression of these two proteins. We also found a negative correlation between the size of breast carcinomas and the expression of nm23-H1, a higher proportion of nm23-H1-positive carcinomas in the group of erbB-2-negative, p53-negative carcinomas and a higher proportion of nm23-H1-positive carcinomas in the group of malignant lesions with negative axillary lymph nodes. Our results support the hypothesis that in women with breast cancer the expression of nm23-H1 gene may contribute to more favorable phenotype. We also showed that some changes found in malignant breast tumors such as the presence of mutated p53 protein and the expression of erbB-2 protein may be found in benign lesions as well.

The consequences of insulin-like growth factors/receptors dysfunction in lung cancer.

The aim of this study was to investigate the consequences of insulin-like growth factors (IGF) and IGF receptor dysfunction in lung carcinomas. A correlation between increased expression (at mRNA and protein levels) for IGF-1 and IGF-1R and decreased apoptosis were found in large-cell carcinomas and adenocarcinomas. In 40% of informative adenocarcinomas expressing the highest values of IGF-2 and Ki-67 proteins, M6P/IGF-2R gene had LOH at one allele and a mutation in another allele. All four squamous cell carcinoma samples expressed LOH/mutation in the M6P/IGF-2R gene. The alphaIR3 strongly diminished proliferation and increased apoptosis in cultures established from squamous cell carcinomas overexpressing IGF-2 and IGF-1R. Telomerase activity was assessed in four squamous cell carcinomas. Cell treatment with IGF-1 increased telomerase activity. The opposite was observed when the cells were treated with alphaIR3, which inhibits the activity of IGF-1 receptors. Our findings suggest that disruption of the IGF/IGF receptors axis is involved in lung cancer formation.

Insulin-like growth factor family and combined antisense approach in therapy of lung carcinoma.

BACKGROUND: Perturbation in a level of any peptide from insulin-like growth factor (IGF) family (ligands, receptors, and binding proteins) seems to be implicated in lung cancer formation; IGF ligands and IGF-I receptor through their mitogenic and anti-apoptotic action, and the mannose 6-phosphate/insulin-like growth factor II receptor (M6-P/IGF-IIR) possibly as a tumor suppressor. MATERIALS AND METHODS: To determine the identity, role, and mutual relationship of IGFs in lung cancer growth and maintenance, we examined IGF's gene (by RT-PCR) and protein (by immunohistochemistry) expression in 69 human lung carcinoma tissues. We also examined IGF-I receptor numbers (Scatchard analysis) and IGF-II production and release (by Western blot) in IGF-II/IGF-IR mRNA positive and negative lung carcinomas. Finally, the potential role of IGF-IR and IGF-II as growth promoting factors in lung cancer was studied using antisense oligodeoxynucleotides that specifically inhibit IGF-IR and IGF-II mRNA. RESULTS: Thirty-two tumors were positive for IGF-I, 39 for IGF-II, 48 for IGF-IR, and 35 for IGFBP-4 mRNA. Seventeen tumors were concomitantly positive for all four IGFs, whereas 34 were positive for IGF-II, IGF-IR, and IGFBP-4 mRNA. An elevated amount of IGF-II peptide was secreted into the growth medium of cell cultures established from five different IGF-II/IGF-IR mRNA positive lung cancer tissues. The cells also expressed elevated numbers of IGF-IR. Nine IGF-II-negative and 19 IGF-II-positive lung cancers of different stages were selected, and M6-P/ IGF-II receptor was determined immunohistochemically. Most of the IGF-II-negative tumors were strongly positive for M6-P/IGF-IIR. IGF-II-positive tumors were mostly negative for M6-P/IGF-II receptors. Antisense oligodeoxynucleotides to IGF-II significantly inhibited, by 25-60%, the in vitro growth of all six lung cancer cell lines. However, the best results (growth inhibition of up to 80%) were achieved with concomitant antisense treatment (to IGF-IR and IGF-II). CONCLUSION: Our data suggest that lung cancer cells produce IGF-IR and IGF-II, which in turn stimulates their proliferation by autocrine mechanism. Cancer cell proliferation can be abrogated or alleviated by blocking the mRNA activity of these genes indicating that an antisense approach may represent an effective and practical cancer gene therapy strategy.