Integrin α3ß1-CD151 complex regulates dimerization of ErbB2 via RhoA.

Integrin α3ß1 regulates adhesive interactions of cells with laminins and have a critical role in adhesion-dependent cellular responses. Here, we examined the role of α3ß1-integrin in ErbB2-dependent proliferation of breast cancer cells in three-dimensional laminin-rich extracellular matrix (3D lr-ECM). Depletion of α3ß1 in ErbB2-overexpressing breast cancer cells suppressed growth and restore cell polarity in 3D lr-ECM. The phenotype of α3ß1-depleted cells was reproduced upon depletion of tetraspanin CD151 and mirrored that of the cells treated with Herceptin, an established ErbB2 antagonist. Breast cancer cells expressing the α3ß1-CD151 complex have higher steady-state phosphorylation of ErbB2 and show enhanced dimerization of the protein when compared with α3ß1-/CD151-depleted cells. Furthermore, Herceptin-dependent dephosphorylation of ErbB2 was only observed in α3ß1-CD151-expressing cells. Importantly, the inhibitory activity of Herceptin was more pronounced when cells expressed both α3ß1 and CD151. We also found that the level of active RhoA was increased in α3ß1- and CD151-depleted cells and that Rho controls dimerization of ErbB2. Expression of α3ß1 alone did not have significant prognostic value in patients with invasive ductal carcinoma of the breast. However, expression of α3ß1 in combination with CD151 represented a more stringent indicator of poor survival than CD151 alone. Taken together, these results demonstrate that the α3ß1-CD151 complex has a critical regulatory role in ErbB2-dependent signalling and thereby may be involved in breast cancer progression.

Association of loss of heterozygosity with shorter survival in primary glioblastoma patients.

Loss of heterozygosity (LOH) co-deletion 1p/19q, MGMT promoter methylation and/or IDH1 mutation generally signify a better prognosis for patients with glioma. However, the influence of 1p/19q co-deletion and the LOH on other chromosomes in primary glioblastoma on survival is still debatable. The aim of our study was to identify LOH on chromosomes 1p, 19q, 9p, 10q, 13q, and 17p, and evaluate their impact either alone or 1p/19q co-deletion or by groups of LOH on the overall survival of 42 primary glioblastoma patients without an oligodendroglial component. These patients were additionally molecularly characterized for EGFR amplification, IDH1 mutations and TP53 mutations. We assessed their influence on the overall survival of glioblastoma patients. LOH in at least one of the loci on all examined chromosomes was detected in 65% of cases and was significantly associated with shorter overall survival (hazard ratio 3.07; 95% CI: 1.29-7.31, p = 0.006). 1p/19q co-deletion was infrequent (7.14%) and had no impact on overall survival. Our results indicate that in primary glioblastoma a specific LOH group analysis may be important for the prognosis. LOH 1p/19q co-deletion is rare in glioblastoma without an oligodendroglial component and has no impact on patient survival.

Loss of CD151/Tspan24 from the complex with integrin α3ß1 in invasive front of the tumour is a negative predictor of disease-free survival in oral squamous cell carcinoma.

OBJECTIVES: The study aimed to assess the role of CD151-integrin α3ß1 (INGA3) complex as a potential prognostic indicator in OSCC and to examine whether mapping of its expression in the invasive front separately from that in the rest of the tumour would have an impact on the predictive value of the results. CD151/INGA3 profiles were compared with that of EGFR. MATERIALS AND METHODS: Protein distributions were analysed either in the whole tumour (W) or separately, (i) the main tumour mass (TU) and (ii) the invasive front (IF) in 83 OSCC samples using immunohistochemistry. RESULTS AND CONCLUSION: There was no statistical association between any of the proteins scored in W and clinicopathologic features or patient survival. When examined separately, significant associations were shown for (i) CD151 and EGFR in TU (p=0.036) and (ii) tumour grade and EGFR in both TU (p=0.045) and IF (p=0.030). INGA3 was present predominantly in the tumour-host interface, significantly stronger in IF than TU (p=0.021). An association with 5-year disease-free survival was close to significant for INGA3 (TU and IF) (p=0.050) but not the CD151/INGA3 complex. Expression of CD151/INGA3 at the IF might reflect tumour behaviour pertinent to patient outcome.

Maspin and Nm23-H1 expression in colorectal cancer.

The aim of the study was to analyze the expression of Nm23-H1 and maspin proteins in a series of colorectal adenocarcinoma and to assess their applicability as prognostic factors in this type of cancer. 102 specimens of colorectal carcinoma were analyzed by immunohistochemistry with the use of anti-Nm23-H1 and anti-maspin monoclonal antibodies. Cytoplasmic expression of Nm23-H1 and maspin was found in 90 of all investigated cases. In 60 cases maspin protein was found also in nucleus. Medium/high Nm23-H1 cytoplasmic expression level was associated with tubular type of adenocarcinoma with deeper invasion of cancer into intestinal wall (T3, T4) and presence of vascular invasion. Medium/high expression level of maspin was connected uniformly with bad prognostic features: low differentiation of tumors (G3), deeper invasion of cancer (T3, T4) presence of nodular and distant metastases, higher Astler-Coller stage (C1, C2, D) and presence of vascular invasion. No statistically significant associations between presence of nuclear maspin expression and any clinicopatological and biological features were stated. Cytoplasmic medium/high expression level of maspin but no Nm23-H1 and no presence of maspin nuclear expression was found as independent bad prognostic factor in the investigated group of patients. Measurement of level and cellular pattern of maspin expression could be valuable for predicting disease course in patients suffering from colorectal cancer.

Astonishingly rapid growth of malignant cystosarcoma phyllodes tumor in a pregnant woman--a case report.

There are very few reports concerning the presence of malignant cystosarcoma phyllodes (CSP) in breasts of pregnant women. In the hereby described case, a 28-year-old woman presented in our department with huge (18 x 11 x 8 cm) tumor of left breast, 2 weeks after labor. The patient discovered a tumor in 34th week of pregnancy, 6 weeks before labor. Histopatholgic examination of excised tumor revealed the presence of malignant CSP tumor. Simple mastectomy was proposed to patient as a best treatment modality. However, the patient refused. She underwent excision of tumor bed (2-cm tumor-free margin was achieved). Despite insufficient treatment, she remains free of disease 20 months after the wide excision of breast malignancy. It is not known how pregnancy influences prognosis of patients with malignant CSP. Lack of such information prompted us to describe the clinical course of our patient.

Evaluation of oestrogen receptor expression in breast cancer by quantification of mRNA.

AIMS: cDNA microarrays have subclassified breast carcinomas into molecular subtypes with oestrogen receptor-alpha (ER) gene expression as a main marker. The aim was to compare ER expression in 97 patients with operable breast cancer estimated by real-time reverse transcriptase-polymerase chain reaction (real-time RT-PCR) and by routine immunohistochemistry, and to determine which method was reliable for molecular subtyping in relation to basal-type keratins and HER2 gene expression. METHODS AND RESULTS: Frozen tumour samples were analysed by real-time RT-PCR for the expression of ER, HER2, keratin 5 and keratin 17 genes. In a group of 27 tumours with a low level of ER mRNA (<1.00), there were eight ER+ cases as assessed by immunohistochemistry, and of 70 cases with a high level of ER mRNA (>or=1.00), 26 were ER- by immunohistochemistry (P = 0.003). Lack of prognostic relevance of ER mRNA level was demonstrated, whereas assessment by immunohistochemistry was related to clinical outcome. Expression of basal keratins and HER2 genes differed significantly between ER+ and ER- tumours based on immunohistochemistry, but not on mRNA level. CONCLUSIONS: These results throw doubt on the assessment of ER mRNA as a key factor in the molecular distinction between breast tumours.

The infrequent simultaneous genetic alterations in glioblastoma multiforme (LOH 10, 17, 19q, TP53 mutation and EGFR amplification) with short clinical course.

We described the case of an unusual, complex genetic alteration in 57 year-old male patient with glioblastoma multiforme (GBM) with short survival (6 and half months). Alterations consisted of p53 mutation, LOH 10, LOH 17, LOH 19q and EGFR amplification. LOH1p, LOH 9 and LOH 13 were negative. Immunohistochemical study did not correlate with molecular results. The overexpression of TP53 protein and RB protein was detected only in small percentage of cells and interestingly the overexpression of EGFR was present only focally. Immnunostainings for PTEN, P16, PI3-K were negative. Additionally, we observed an overexpression of IGFB2 protein. This case indicates the accumulation of molecular changes in glioblastoma multiforme in patient with short survival.

Cyclin E expression in breast cancer correlates with negative steroid receptor status, HER2 expression, tumor grade and proliferation.

The main objective of this retrospective study was to investigate relations between cyclin E and pathoclinical factors in patients with operable breast cancer. Expression of cyclin E was analyzed by immunohistochemistry in specimens of invasive ductal breast cancer tissue obtained from 189 women during radical mastectomy. Overall, 110 tumor samples were regarded to be cyclin E positive. Cyclin E expression was more often seen in tumors with: negative steroid receptor status (p<0.0001), higher proliferative index (p=0.0014), higher tumor grade (p=0.0017), and presence of HER2 (p=0.0171). With a median follow-up of 58 months, expression of cyclin E together with negative steroid receptor status determined poor prognosis with a 5-year cancer-specific survival rate of 58%. It differed significantly from a survival curve of cyclin E negative and steroid receptor positive patients (87%, p=0.0005). No significant difference was observed in comparison with survival of cyclin E positive and steroid receptor positive patients (68%, p=0.221). We demonstrated that cyclin E expression in breast cancer cells was associated with negative steroid receptor status, HER2 presence, higher tumor grade and higher proliferation index. Expression of cyclin E together with lack of steroid receptors determined poor prognosis.

WWOX--the FRA16D cancer gene: expression correlation with breast cancer progression and prognosis.

AIMS: WWOX is a tumour suppressor gene involved in various tumours including breast cancer. High chromosomal abnormalities in a genomic region spanned by WWOX are associated with the fact that this gene covers approximately 1 million base pairs of the second most affected among common chromosomal fragile sites FRA16D. We evaluated WWOX expression levels in breast cancer samples in association with diagnostics-prognostics markers. METHODS: We performed quantitative real-time RT-PCR to analyse levels of expression of WWOX in 132 cases of breast cancer. We evaluated the relationship between WWOX mRNA levels, clinico-pathological factors, expression of aberrant WWOXDelta6-8 mRNA and other cancer related genes. RESULTS: Expression of WWOX was higher in patients younger than 50 years old, in ER and PR positive tumours vs negative for those receptors and tumours without lymph node metastasis vs LN+. WWOX mRNA levels were also higher in tumours with higher apoptotic index (Bcl2/Bax ratio). Negative associations were found between WWOX expression and cytokeratins 5/6 and 17 (P<0.05). High level expression of WWOX was also associated with better disease free survival. Presence of WWOXDelta6-8 transcripts were accompanied with lower WWOX wild type mRNA level. CONCLUSIONS: Reduced WWOX expression commonly observed in various neoplasias in cases of breast cancer is associated with markers of bad prognosis. Our findings reveal additional evidence that WWOX may be involved in steroid (estrogens) metabolism and signaling pathways. WWOX can be considered as a new target for gene therapy development due to the association of high WWOX expression with improved disease free survival.

[Pharmacological action of paclitaxel]. / Farmakologia kliniczna paklitakselu.

Paclitaxel (taxol, Tax) is a novel plant product isolated from the Pacific yew (Taxus brevifolia). It has a unique mechanism of action, because it induces very stable and dysfunctional microtubules. Paclitaxel has a broad spectrum of antineoplastic activity. It has been successfully used in the treatment of ovarian cancer, metastatic breast cancer, lung cancer, carcinoma of the head and neck, malignant melanoma and other human neoplasms. Tax has response rates of 20-36% in patients with refractory ovarian cancer. Toxic effects include myelosuppresion, hypersensitivity reactions, peripheral neuropathy, cardiac disturbances, alopecia. However, studies evaluating the drug still are ongoing paclitaxel seems to be one of the most promising antineoplastic agents.

Activity of 2-chlorodeoxyadenosine (Cladribine) in 2-hour intravenous infusion in 94 previously treated patients with low grade non-Hodgkin's lymphoma.

The purpose of our study was to determine the efficacy of 2-chlorodeoxyadenosine (2-CdA) administered in 2-hour intravenous infusions in previously treated patients with low grade non-Hodgkin's lymphoma (LGNHL). We treated 94 LGNHL patients with 2-CdA at a dosage of 0.12 mg/kg/24h in 2-hour intravenous infusion for 5 consecutive days. The treatment consisted of from 1 to 7 courses (median 3), repeated usually at monthly intervals. All patients were refractory to or relapsed after standard chemotherapy. Of these 94 patients 78 (83%) had clinical stage IV of the disease. Complete response (CR) was obtained in 12 (12.8%) and partial response (PR) in 36 (38.3%) giving an overall response rate of 51.1%. In 12 (12.8%) grade 4 thrombocytopenia with haemorrhagic diathesis was noted, grade 4 neutropenia was observed in 12 (12.8%) and infections complicated the course of treatment in 38 (40.4%) patients. 2-CdA treatment was the cause of death of 3 patients. The results of our study show that 2-CdA given in 2-hour infusions is an effective agent in advanced, heavily pretreated patients with LGNHL.

Metoclopramide and nitrendipine are unable to enhance cytotoxicity of mitoxantrone in sensitive and resistant murine leukemias in vivo.

The influence of metoclopramide and nitrendipine on the antitumor action of mitoxantrone (MX) in murine leukemias L1210 and P388 was investigated. These agents were administered in combination with MX to mice bearing sensitive or resistant leukemias. Mitoxantrone resistant P388 was developed in vivo by repeated passages of the tumor to animals treated with suboptimal dose of the drug. Survival time of mice receiving combined therapy were not prolonged as compared with animals treated with MX alone. Both metoclopramide and nitrendipine were unable to overcome MX resistance even partially.

Intermittent 2-hour intravenous infusions of 2-chlorodeoxyadenosine in the treatment of 110 patients with refractory or previously untreated B-cell chronic lymphocytic leukemia.

The purpose of our study was to determine the effectiveness of 2-CdA in 2-hour intravenous infusions in the treatment of B-CLL. One hundred and ten patients with B-CLL received 1 to 10 courses of 2-CdA (median 2.5) at a dosage of 0.12 mg/kg daily for 5 consecutive days. Eighteen of them were untreated and 92 relapsed or became refractory to previous therapeutic modalities. Complete remission (CR) was achieved in 8 (7.3%) and partial remission (PR) in 35 patients (31.8%) giving an overall response rate of 39.1%. In 3 patients, cross-resistance to fludarabine was noticed. Toxic effects of 2-CdA were more frequently observed in previously treated patients. Hemorrhagic complications due to drug-induced thrombocytopenia were noticed in 25 (22.7%) and severe infections including sepsis in 14 (12.7%) patients.