Analytical Cellular Pathology - Volume 31, issue 6

Authors: Kaufmann, Simone | Kuphal, Silke | Schubert, Thomas | Bosserhoff, Anja K.

Article Type: Research Article

Abstract: Background: Malignant melanoma cells are known to have altered expression of genes supporting proliferation and invasion, however, the expression of molecules of the Netrin family of repellent factors has not been analyzed in melanomas until now. Results: Here, we show that Netrin-1 expression is strongly induced in melanoma cells compared to melanocytes in vivo and in vitro controlled at the transcriptional level via ETS-1. In addition, the expression of the netrin receptor UNC5B was induced and that of UNC5C was reduced in the tumor cells. In order to determine the functional relevance of Netrin-1 expression in malignant melanoma, Netrin …expression in melanoma cells was reduced by siRNA attempts and primary human melanocytes were treated with recombinant Netrin-1. The cells showed no changes in proliferation or apoptosis, however, a strong reduction of migratory properties was observed in the melanoma cells after reduction of Netrin expression whereas melanocyte migration was strongly induced by treatment with Netrin. Conclusion: Our study suggests that Netrin-1 promotes melanoma cell invasion and migration and therefore has an important role in the progression of malignant melanoma. Show more

Keywords: Malignant melanoma, migration, repellent factor, Netrin

DOI: 10.3233/CLO-2009-0491

Citation: Analytical Cellular Pathology, vol. 31, no. 6, pp. 415-422, 2009

Authors: Valente, Guido | Nicotra, Giuseppina | Arrondini, Marisa | Castino, Roberta | Capparuccia, Lorena | Prat, Maria | Kerim, Simonetta | Tamagnone, Luca | Isidoro, Ciro

Article Type: Research Article

Abstract: Background: Plex-B1, the receptor of Sema4D, has been implicated in tumour growth, angiogenesis and metastasis. The binding of Sema4D to Plex-B1 can trigger the activation of Met tyrosine kinase, thereby promoting cell dissociation and invasive growth. We tested the hypothesis that the expression of Plex-B1, either alone or in association with Met, can be of predictive value for tumour progression. Methods: The expression and distribution of Plex-B1 and Met were investigated by immunohistochemistry and immunofluorescence in 50 human neoplasias originating in the breast and ovary, and correlated with clinical–pathological data at diagnosis. Results: Plex-B1 and Met were individually …expressed in 14% and in 24% of the tumours, respectively. Plex-B1 and Met were co-expressed in 24/50 cases (48%), and in the majority of these (83%) Met was tyrosine phosphorylated. The expression of Plex-B1 or Met alone showed no significant correlation with tumour aggressiveness, whereas advanced stage tumours (III–IV) frequently showed Plex-B1–Met double-positive (9/13). Tumours co-expressing Plex-B1 and Met were characterised by worse grading and higher incidence of lymph node metastases. Out of 22 tumours with lymph node metastases, as many as 19 were Plex-B1 and Met double-positive (p=0.0008), and 17 expressed phosphorylated Met (p=0.002). Conclusion: Plex-B1 assumes a predictive value for unfavourable outcome when co-expressed with Met. Show more

Keywords: Plexin-B1, Met, CD100, tumour prognosis, lymph node metastases

DOI: 10.3233/CLO-2009-0504

Citation: Analytical Cellular Pathology, vol. 31, no. 6, pp. 423-436, 2009

Authors: Hassan, Mohamed | Feyen, Oliver | Grinstein, Edgar

Article Type: Research Article

Abstract: Renal cell carcinoma (RCC) is a prototype of a chemo refractory tumour. It remains the most lethal of the common urologic cancers and is highly resistant to conventional therapy. Here, we confirmed the efficiency of anti-Fas monoclonal antibody (CH11) as alternative therapeutic approach for the treatment of RCC and investigated the molecular mechanism(s), whereby CH11 induces apoptosis of RCC cells. The present study shows an essential role for apoptosis signal-regulating kinase 1 (ASK1), together with both c-jun-N-terminal kinase (JNK) and p38 pathways, and caspase-8 in this process. Furthermore, CH11-dependent induction of the ASK1–JNK/p38 pathways was found to activate the transcription …factors AP-1 and ATF-2, and FADD-caspase-8-Bid signalling, resulting in the translocation of both Bax and Bak proteins, and subsequently mitochondrial dysregulation that is characterized by the loss of mitochondrial membrane potential (ΔΨm ), cytochrome c release and cleavage of caspase-9, caspase-3 and PARP. Thus, the described molecular mechanisms of CH11-induced apoptosis suggest the reliability of Fas activation as an alternative therapeutic approach for the treatment of patients with advanced renal cell carcinoma. Show more

Keywords: Apoptosis, RCC, CH11, ASK1, MAP kinase

DOI: 10.3233/CLO-2009-0488

Citation: Analytical Cellular Pathology, vol. 31, no. 6, pp. 437-456, 2009

Authors: Zatelli, Maria Chiara | Molè, Daniela | Tagliati, Federico | Minoia, Mariella | Ambrosio, Maria Rosaria | Uberti, Ettore degli

Article Type: Research Article

Abstract: Background: Breast cancer cells can develop chemoresistance after prolonged exposure to cytotoxic drugs due to expression of the multi drug resistance (MDR) 1 gene. Type 2 cyclo-oxygenase (COX-2) inhibitors reverse the chemoresistance phenotype of a medullary thyroid carcinoma cell line, TT, and of a breast cancer cell line, MCF7, by inhibiting MDR1 expression and P-gp function. Aim: investigate the role of prostaglandin (PG) in modulating chemoresistance in MCF7 cells and to explore the involved intracellular mechanisms. Methods: native and chemoresistant MCF7 cells were treated with PGH2 and resistance to Doxorubicin was tested in the presence or absence …of COX-2 inhibitors. Results: PGH2 restores resistance to the cytotoxic effects of Doxo, with concomitant nuclear translocation of the transcription factor NF-κB. Conclusions: COX-2 inhibitors prevent chemoresistance development in breast cancer cells by inhibiting P-gp expression and function by a mechanism that involves PGH2 generation and NF-κB activation. Show more

Keywords: PGH_2, P-gp, COX-2 inhibitors, NF-κB, chemoresistance, breast cancer

DOI: 10.3233/CLO-2009-0490

Citation: Analytical Cellular Pathology, vol. 31, no. 6, pp. 457-465, 2009

Authors: Cummings, Michele | Iremonger, James | Green, Caroline A. | Shaaban, Abeer M. | Speirs, Valerie

Article Type: Research Article

Abstract: Background: Despite many published studies on ERβ, progress towards understanding its role in breast cancer remains slow. This is largely due to discordant data between mRNA and protein studies as well as failure to take into account the biologically distinct ERβ isoforms and their heterogeneous expression profile. Methods: We compared expression of ERβ1, -2 and -5 genes in HB2 and MCF-7 breast cell lines, primary breast fibroblasts (n=5) and whole tissue and laser microdissected epithelial and stromal cells obtained from 25 human breast tumours. Results: Our study shows that the level of gene expression of ERβ isoforms depends …on the cell population within a given tumour and varies dramatically in different cellular compartments. This has implications for gene expression analyses and could explain some of the contradictory data published to date, rendering “grind and bind” analyses of ERβ uninformative. Conclusion: With the technology now available, we suggest a more refined approach be adopted to help resolve some of the controversy surrounding ERβ. Show more

Keywords: Estrogen receptor β, laser microdissection, breast cancer

DOI: 10.3233/CLO-2009-0495

Citation: Analytical Cellular Pathology, vol. 31, no. 6, pp. 467-473, 2009

Authors: Sitarz, R. | Leguit, R.J. | de Leng, W.W.J. | Morsink, F.H.M. | Polkowski, W.P. | Maciejewski, R. | Offerhaus, G.J.A. | Milne, A.N.

Article Type: Research Article

Abstract: Background: COX-2 and E-cadherin, involved in invasion and metastasis, are molecules critical for gastric carcinogenesis. A relationship between them is documented in non-small cell lung and prostate cancer. We present novel evidence of a relationship between COX-2 and E-cadherin expression in gastric cancer. Methods: Using qPCR and Western blots analysis on celecoxib and PGE2 treated and untreated gastric cancer cell lines derived from tumours of the intestinal type (MKN45, MKN28, AGS3, MKN7) and immunohistochemistry of 178 gastric cancers on tissue microarrays (TMA), we examined the COX-2/E-cadherin relationship. Results: Down-regulation of COX-2 by celecoxib led to up-regulation of E-cadherin …mRNA and protein levels in conventional gastric cancer cell lines, whereas expression was down regulated in the early-onset gastric cancer (EOGC) cell line. Immunohistochemistry on TMAs of 178 gastric cancers showed no correlation between COX-2 and E-cadherin expression in the conventional or early gastric cancer groups. Conclusion: The results suggest that COX-2 has an impact on transcriptional regulation of E-cadherin in gastric cancer and our findings further highlight the intriguing nature of EOGCs which appear to have a molecular phenotype distinct from conventional gastric cancer. In addition, our findings also suggest that reduction of COX-2 using nonsteroidal anti-inflammatory drugs in gastric cancer chemoprevention may only be relevant for older patients. Show more

Keywords: COX-2, E-cadherin, early-onset gastric cancer, CDH1 mutation, celecoxib

DOI: 10.3233/CLO-2009-0496

Citation: Analytical Cellular Pathology, vol. 31, no. 6, pp. 475-485, 2009

Authors: Sharma, Gayatri | Mirza, Sameer | Yang, Yi-Hsin | Parshad, Rajinder | Hazrah, Priya | Datta Gupta, Siddartha | Ralhan, Ranju

Article Type: Research Article

Abstract: Background: Methylation-mediated suppression of detoxification, DNA repair and tumor suppressor genes has been implicated in cancer development. This study was designed to investigate the impact of concurrent methylation of multiple genes in breast tumors on disease prognosis. Methods: Methylation specific PCR was carried out to analyze the methylation status of seven genes in archived breast tissues and determine the effect of aberrant methylation of multiple genes on disease prognosis and patients' survival. Results: Promoter hypermethylation was observed in PRB 67%, ERα 64%, RASSF1A 63%, p16INK4A 51%, RARβ2 22%, GSTP1 25% and BRCA1 27% of the breast cancers, respectively. …Concurrent methylation of BRCA1, ERα, GSTP1 and RARβ2, was observed in a large proportion of breast cancers analyzed, suggesting that these genes do not appear to be methylated alone. Patients with high methylation indices had poor prognosis (p<0.001, Hazards ratio=14.58). Cox regression analysis showed RARβ2 promoter methylation to be an independent important determinant of breast cancer prognosis. Conclusion: Our results suggest that methylation of multiple genes plays an important role in prognosis of breast cancer. Our study not only describes the association of methylation mediated silencing of multiple genes with the severity of disease, but also drives to speculate the molecular crosstalk between genes or genetic pathways regulated by them individually. Show more

Keywords: Breast cancer, methylation, prognosis, RARβ, GSTP1, BRCA1

DOI: 10.3233/CLO-2009-0507

Citation: Analytical Cellular Pathology, vol. 31, no. 6, pp. 487-500, 2009

Authors: Albregtsen, Fritz

Article Type: Other

DOI: 10.3233/CLO-2009-0506

Citation: Analytical Cellular Pathology, vol. 31, no. 6, pp. 501-501, 2009

Authors: Metze, Konradin | Lorand-Metze, Irene | Leite, Neucimar J | Adam, Randall L.

Article Type: Other

DOI: 10.3233/CLO-2009-0505

Citation: Analytical Cellular Pathology, vol. 31, no. 6, pp. 503-504, 2009

Article Type: Other

Citation: Analytical Cellular Pathology, vol. 31, no. 6, pp. 505-510, 2009