Analytical Cellular Pathology - Volume 28, issue 4

Authors: Fijneman, Remond J.A.

Article Type: Research Article

Citation: Analytical Cellular Pathology, vol. 28, no. 4, pp. 125-126, 2006

Authors: Koo, V. | Hamilton, P.W. | Williamson, K.

Article Type: Research Article

Abstract: Non-invasive real time in vivo molecular imaging in small animal models has become the essential bridge between in vitro data and their translation into clinical applications. The tremendous development and technological progress, such as tumour modelling, monitoring of tumour growth and detection of metastasis, has facilitated translational drug development. This has added to our knowledge on carcinogenesis. The modalities that are commonly used include Magnetic Resonance Imaging (MRI), Computed Tomography (CT), Positron Emission Tomography (PET), bioluminescence imaging, fluorescence imaging and multi-modality imaging systems. The ability to obtain multiple images longitudinally provides reliable information whilst reducing animal numbers. As yet there …is no one modality that is ideal for all experimental studies. This review outlines the instrumentation available together with corresponding applications reported in the literature with particular emphasis on cancer research. Advantages and limitations to current imaging technology are discussed and the issues concerning small animal care during imaging are highlighted. Show more

Keywords: Animal model, non-invasive, in vivo, imaging, CT, PET, MRI, bioluminescence, fluorescence

Citation: Analytical Cellular Pathology, vol. 28, no. 4, pp. 127-139, 2006

Authors: Mesker, Wilma E. | Vrolijk, Hans | Sloos, Willem C.R. | Tollenaar, Rob A.E.M. | Tanke, Hans J.

Article Type: Research Article

Abstract: The presence of tumor cells in bone marrow, peripheral blood and lymph nodes has proven its clinical and prognostic value. Since the frequency of these cells in bone marrow and blood is sometimes as low as 1 per million and due to the fact that for the analysis of lymph nodes many sectioning levels have to be analyzed, automated imaging devices have been suggested as an useful alternative to conventional manual screening of specimens. The aim of this paper is to review the performance of current equipment that is commercially available, based on literature published so far. Requirements for introducing …this equipment for routine clinical practice are discussed. Show more

Keywords: Disseminated tumor cells, micrometastases, immunohistochemistry, image analysis, automated devices (microscopy)

Citation: Analytical Cellular Pathology, vol. 28, no. 4, pp. 141-150, 2006

Authors: Moerland, Elna | van Hezik, Rens L.H.P.M. | van der Aa, Toine C.J.M. | van Beek, Mike W.P.M. | van den Brule, Adriaan J.C.

Article Type: Research Article

Abstract: In this study the detection of HER2 gene amplification was evaluated using Fluorescence In Situ Hybridization (FISH; PathVysion) in comparison with Multiplex Ligation-dependent Probe Amplification (MLPA), a PCR based technique. These two methods were evaluated on a series of 46 formalin fixed paraffin embedded breast carcinomas, previously tested for protein overexpression by HercepTest (grouped into Hercep 1+, 2+ and 3+). HER2 gene amplification (ratio ≥2.0) by FISH was found in 9/10, 10/30 and 0/6 in IHC 3+, 2+ and 1+/0 cases, respectively. Digitalized automated spot counting performed with recently developed CW4000 CytoFISH software was 100% concordant with manual FISH scoring. …Using MLPA 18/46 samples showed a clear HER2 amplification. Comparing MLPA and IHC showed the same results as for FISH and IHC. All but one FISH positive cases (18/19) were confirmed by MLPA for the presence of the gene amplification. The overall concordance of detection of Her2 gene amplification by FISH and MLPA was 98% (45/46). Furthermore, both the level of amplification and equivocal results correlated well between both methods. In conclusion, MLPA is a reliable and reproducible technique and can be used as an either alternative or additional test to determine HER2 status in breast carcinomas. Show more

Keywords: HER2, amplification, MLPA, FISH, automatic, quantification, spot counting

Citation: Analytical Cellular Pathology, vol. 28, no. 4, pp. 151-159, 2006

Authors: Calistri, Daniele | Rengucci, Claudia | Seymour, Ian | Leonardi, Elena | Truini, Mauro | Malacarne, Davide | Castagnola, Patrizio | Giaretti, Walter

Article Type: Research Article

Abstract: Background: The origin and mechanisms of chromosomal instability are still widely unknown. We previously investigated a limited number of human sporadic colorectal cancers (CRCs) and observed a statistically different occurrence of KRAS and p53 mutations among predetermined subgroups of tumors with different degrees of DNA aneuploidy. The aim of the present study was to further verify these observations by including BRAF gene analysis and by investigating a larger series of cases subdivided into Dukes' stages A to D to reconstruct some form of chronological modulation for events during CRC progression. Methods: KRAS, p53, BRAF mutations and flow cytometric DNA Index …were evaluated by established techniques in a series of 135 human sporadic CRCs. Results: p53, KRAS and BRAF mutations were found in 39%, 34%, and 4% of tumors, respectively. The frequency of p53 mutations increased from 15% for stage A to 48% for stage D and was highest in near-diploid (DI < 1.4 and DI ≠ 1) and high-aneuploid (DI > 1.6) tumors. A similar correlation between gene mutations and DI values was observed for KRAS. The simultaneous presence of KRAS and p53 mutations was observed in only 11% of cases. Moreover, the co-occurrence of p53 and KRAS mutations was only observed in near-diploid and high-aneuploid tumors. Conclusion: Our findings suggest that KRAS and p53 gene mutations, which are rarely simultaneous and are associated with specific DI aneuploid values, do not represent a synergistic evolutionary pathway but may influence mechanisms of chromosomal instability. Show more

Keywords: Gene mutations, aneuploidy, colorectal cancer

Citation: Analytical Cellular Pathology, vol. 28, no. 4, pp. 161-166, 2006

Authors: Knösel, Thomas | Emde, Valeska | Schlüns, Karsten | Schlag, Peter Michael | Dietel, Manfred | Petersen, Iver

Article Type: Research Article

Abstract: Background and aims: Recent cDNA expression profiling analyses indicate that within specific organ cancers Cytokeratins (CKs) dysregulation may identify subgroups with distinct biological phenotypes. Our objectives in this study were (1) to test whether cytokeratins were also distinct on the protein level, (2) to evaluate these biomarkers in a series of well-characterised CRCs, (3) to apply hierarchical cluster analysis to immunohistochemical data. Methods: Tissue microarrays (TMA) comprising 468 CRC specimens from 203 patients were constructed to evaluate CK5, CK7, CK8, CK13, CK14, CK16, CK17, CK18, CK19 and CK20. In total, 2919 samples were analyzed. Results: Unsupervised hierarchical clustering discovered subgroups …represented by reduced CK8 and CK20 expression, that differed by a shorter patients survival. The evaluation of the specific biomarkers by Kaplan–Meier analysis showed that reduced CK8 expression (p<0.01) was significantly associated with shorter patients' survival, but was not an independent factor correlated with tumour stage (pT), grading (G) and nodal stage (pN). Conclusions: Reduced coexpression of CK8 and CK20 may indicate an epithelial-mesenchymal transition (EMT) representing an important step in the development of more aggressive CRCs. In addition, multiplex analysis of TMAs together with immunohistochemistry (IHC) supplemented by hierarchical clustering are a useful, promising and very powerful tool for the identification of tumour subgroups with diagnostic and prognostic signatures. Show more

Keywords: Cytokeratins, colorectal cancer (CRC), tissue microarray (TMA), prognosis, hierarchical clustering, epithelial–mesenchymal-transition (EMT)

Citation: Analytical Cellular Pathology, vol. 28, no. 4, pp. 167-175, 2006

Authors: Smirnova, Irina S. | Aksenov, Nikolai D. | Kashuba, Elena V. | Payakurel, Pawan | Grabovetsky, Vadim V. | Zaberezhny, Alexei D. | Vonsky, Maxim S. | Buchinska, Lubov | Biberfeld, Peter | Hinkula, Jorma | Isaguliants, Maria G.

Article Type: Research Article

Abstract: The oncogenic potential of hepatitis C virus (HCV) core protein has been demonstrated, but the precise mechanism of cell transformation triggered by HCV core is still unclear. This study shows that constitutive expression of HCV core protein (core) in NIH 3T3 murine fibroblasts triggers malignant transformation. At the preneoplastic stage, clones that expressed HCV core constitutively demonstrated genomic instability seen as disruption of the mitotic spindle cell checkpoint leading to increased ploidy. Transformation was completed by the loss of DNA and resistance to apoptosis induced by serum starvation. Simultaneously, cells acquired a capacity for anchorage independent growth and absence of …contact inhibition. Inoculation of these transformed cells into severe combined immune deficiency (SCID) mice led to formation of solid core-expressing tumors. Transformation and tumorigenicity of core-expressing cell lines coincided with a 5- to 10-fold repression of endogenous p53 transactivation. Thus, long-term HCV core expression alone is sufficient for complete transformation of immortal fibroblasts that can then induce tumors in a susceptible host. This data suggests that malignant transformation by HCV core may occur through primary stress, induction of genomic instability, and further HCV core-induced rescue of surviving mutated cells. Show more

Keywords: Hepatitis C virus, core protein, constitutive expression, fibroblast, mitotic spindle cell checkpoint, genomic instability, malignant transformation, p53, fibrosarcoma

Citation: Analytical Cellular Pathology, vol. 28, no. 4, pp. 177-190, 2006