Analytical Cellular Pathology - Volume 28, issue 5-6

Authors: Kortenhorst, Madeleine S.Q. | Carducci, Michael A. | Shabbeer, Shabana

Article Type: Research Article

Citation: Analytical Cellular Pathology, vol. 28, no. 5-6, pp. 191-222, 2006

Authors: Mikesch, Jan-Henrik | Schier, Kathrin | Roetger, Antje | Simon, Ronald | Buerger, Horst | Brandt, Burkhard

Article Type: Research Article

Abstract: Decay-accelerating factor (DAF, CD55) is physiologically acting as an inhibitor of the complement system, but is also broadly expressed in malignant tumours. Here DAF seems to exert different functions beyond its immunological role such as e.g. promotion of tumorigenesis, decrease of complement mediated tumor cell lysis, autocrine loops for cell rescue and evasion of apoptosis, neoangiogenesis, invasiveness, cell motility, and metastasis via oncogenic tyrosine kinase pathways and specific seven-span transmembrane receptors (CD97) binding. Therefore, DAF has already become a target for therapy. In this paper we review the role of DAF in human malignancies as described in different basic, diagnostic …and experimental therapeutic studies. Show more

Citation: Analytical Cellular Pathology, vol. 28, no. 5-6, pp. 223-232, 2006

Authors: Nijhuis, Esther R. | Reesink-Peters, Nathalie | Wisman, G. Bea A. | Nijman, Hans W. | van Zanden, Jelmer | Volders, Haukeline | Hollema, Harry | Suurmeijer, Albert J.H. | Schuuring, Ed | van der Zee, Ate G.J.

Article Type: Research Article

Abstract: Although current cytomorphology-based cervical cancer screening has reduced the incidence of cervical cancer, Pap-smears are associated with high false positive and false negative rates. This has spurred the search for new technologies to improve current screening. New methodologies are automation of Pap-smear analysis, addition of new biological or molecular markers to traditional cytology or using these new markers to replace the current screening method. In this overview we will summarize data on cervical cancer epidemiology and etiology and the current cervical cancer screening approach. Available data on new screening approaches, such as quantitative cytochemistry, detection of loss of heterozygosity (LOH) …and hypermethylation analysis will be reviewed. We discuss the potential of these approaches to replace or augment current screening. When available, data on cost–effectiveness of certain approaches will be provided. In short, Human Papillomavirus (HPV) DNA detection stands closest to implementation in nation-wide screening programs of all markers reviewed. However, specificity is low in women aged <35 years and the psychological effects of knowledge of HPV positivity in absence of cervical (pre) malignant disease are important drawbacks. In our opinion the results of large clinical trials should be awaited before proceeding to implement HPV DNA detection. New technologies based on molecular changes associated with cervical carcinogenesis might result in comparable sensitivity, but improved specificity. Hypermethylation analysis is likely to be more objective to identify patients with high grade squamous intra-epithelial lesions (HSIL) or invasive cancer with a higher specificity than current cytomorphology based screening. Show more

Keywords: Biological marker, HPV, methylation, cervical cancer, cervical intraepithelial neoplasia, screening

Citation: Analytical Cellular Pathology, vol. 28, no. 5-6, pp. 233-246, 2006

Authors: Derks, Sarah | Postma, Cindy | Moerkerk, Peter T.M. | van den Bosch, Sandra M. | Carvalho, Beatriz | Hermsen, Mario A.J.A. | Giaretti, Walter | Herman, James G. | Weijenberg, Matty P. | Bruïne, Adriaan P. de | Meijer, Gerrit A. | van Engeland, Manon

Article Type: Research Article

Abstract: Background: Colorectal cancers are characterized by genetic and epigenetic alterations. This study aimed to explore the timing of promoter methylation and relationship with mutations and chromosomal alterations in colorectal carcinogenesis. Methods: In a series of 47 nonprogressed adenomas, 41 progressed adenomas (malignant polyps), 38 colorectal carcinomas and 18 paired normal tissues, we evaluated promoter methylation status of hMLH1, O6 MGMT, APC, p14ARF , p16INK4A , RASSF1A, GATA-4, GATA-5, and CHFR using methylation-specific PCR. Mutation status of TP53, APC and KRAS were studied by p53 immunohistochemistry and sequencing of the APC and KRAS mutation cluster regions. Chromosomal alterations were evaluated by …comparative genomic hybridization. Results: Our data demonstrate that nonprogressed adenomas, progressed adenomas and carcinomas show similar frequencies of promoter methylation for the majority of the genes. Normal tissues showed significantly lower frequencies of promoter methylation of APC, p16INK4A , GATA-4, and GATA-5 (P-values: 0.02, 0.02, 1.1×10−5 and 0.008 respectively). P53 immunopositivity and chromosomal abnormalities occur predominantly in carcinomas (P values: 1.1×10−5 and 4.1×10−10 ). Conclusions: Since promoter methylation was already present in nonprogressed adenomas without chromosomal alterations, we conclude that promoter methylation can be regarded as an early event preceding TP53 mutation and chromosomal abnormalities in colorectal cancer development. Show more

Keywords: Colorectal cancer, promoter methylation, genetic alterations, chromosomal instability

Citation: Analytical Cellular Pathology, vol. 28, no. 5-6, pp. 247-257, 2006

Authors: Lind, Guro E. | Kleivi, Kristine | Meling, Gunn I. | Teixeira, Manuel R. | Thiis-Evensen, Espen | Rognum, Torleiv O. | Lothe, Ragnhild A.

Article Type: Research Article

Abstract: Background: Gene silencing through CpG island hypermethylation is a major mechanism in cancer development. In the present study, we aimed to identify and validate novel target genes inactivated through promoter hypermethylation in colorectal tumor development. Methods: With the use of microarrays, the gene expression profiles of colon cancer cell lines before and after treatment with the demethylating agent 5-aza-2′-deoxycytidine were identified and compared. The expression of the responding genes was compared with microarray expression data of primary colorectal carcinomas. Four of these down-regulated genes were subjected to methylation-specific PCR, bisulphite sequencing, and quantitative gene expression analysis using tumors (n=198), normal …tissues (n=44), and cell lines (n=30). Results: Twenty-one genes with a CpG island in their promoter responded to treatment in cell lines, and were simultaneously down-regulated in primary colorectal carcinomas. Among 20 colon cancer cell lines, hypermethylation was subsequently identified for three of four analyzed genes, ADAMTS1 (85%), CRABP1 (90%), and NR3C1 (35%). For the latter two genes, hypermethylation was significantly associated with absence or reduced gene expression. The methylation status of ADAMTS1, CRABP1, and NR3C1 was further investigated in 116 colorectal carcinomas and adenomas. Twenty-three of 63 (37%), 7/60 (12%), and 2/63 (3%) adenomas, as well as 37/52 (71%), 25/51 (49%), and 13/51 (25%) carcinomas were hypermethylated for the respective genes. These genes were unmethylated in tumors (n=82) from three other organs, prostate, testis, and kidney. Finally, analysis of normal colorectal mucosa demonstrated that the observed promoter hypermethylation was cancer-specific. Conclusion: By using a refined microarray screening approach we present three genes with cancer-specific hypermethylation in colorectal tumors, ADAMTS1, CRABP1, and NR3C1. Show more

Keywords: ADAMTS1, adenomas, bisulphite sequencing, colorectal carcinomas, CRABP1, gene expression, methylation, microarrays, NR3C1, 5-aza-2′deoxycytidine

Citation: Analytical Cellular Pathology, vol. 28, no. 5-6, pp. 259-272, 2006

Authors: Ghadimi, B. Michael | Grade, Marian | Mönkemeyer, Carsten | Kulle, Bettina | Gaedcke, Jochen | Gunawan, Bastian | Langer, Claus | Liersch, Torsten | Becker, Heinz

Article Type: Research Article

Abstract: Background: The prognosis of colorectal cancer patients is to a considerable extent determined by the metastatic potency of the primary tumor. However, despite the fact that liver metastases are the leading cause of death for cancer patients, the molecular basis still remains poorly understood and independent prognostic markers have not been established. Materials and methods: Comparative genomic hybridization (CGH) was used to screen colorectal carcinomas without distant metastases (n=18) and carcinomas synchronously metastatic to the liver (n=18). We aimed to detect distinct chromosomal aberrations indicating a metastatic phenotype. Results and discussion: Metastatic tumors exhibited a significantly (P=0.03) higher ANCA value …(13.8) if compared with non-metastatic cancers (10.0). Furthermore, we observed that losses of chromosomal regions 1p32-ter and 9q33-ter were present at much higher frequencies in metastatic than in non-metastatic cancers, respectively (P=0.02 and 0.04). Conclusion: These data indicate that metastatic tumors may be separated from non-metastatic colorectal cancers based on their genomic profile. Show more

Keywords: Colorectal cancer, comparative genomic hybridization, liver metastases, predictive marker, prognosis

Citation: Analytical Cellular Pathology, vol. 28, no. 5-6, pp. 273-281, 2006

Authors: Carvalho, Beatriz | Buffart, Tineke E. | Reis, Rui M. | Mons, Thomas | Moutinho, Cátia | Silva, Paula | van Grieken, Nicole C.T. | Grabsch, Heike | van de Velde, Cornelis J.H. | Ylstra, Bauke | Meijer, Gerrit A. | Carneiro, Fátima

Article Type: Research Article

Abstract: Gastric cancer is one of the most frequent malignancies in the world. Nonetheless, the knowledge of the molecular events involved in the development of gastric carcinoma is far from complete. One of the hallmarks of gastric cancer is chromosomal instability resulting in abnormal DNA copy number changes throughout the genome. Mixed gastric carcinomas constitute a rare histological entity, containing the two main histological phenotypes (diffuse and intestinal). Very little is known about the underlying mechanisms of phenotypic divergence in these mixed tumours. To the best of our knowledge only E-Cadherin mutations were implicated so far in the divergence of these …tumours and nothing is known about the involvement of chromosome copy number changes in the two divergent histological components. In this study, we compared the DNA copy number changes, in the two different components (diffuse and intestinal) of mixed gastric carcinomas by microarray – comparative genomic hybridisation (array CGH). The analysis of 12 mixed gastric carcinomas showed no significant differences in array CGH profiles between the diffuse and intestinal components of mixed carcinomas. This supports the idea that the phenotypic divergence within mixed gastric carcinomas is not caused by DNA chromosomal aberrations. Show more

Keywords: Mixed gastric carcinoma, chromosomal aberrations, array CGH

Citation: Analytical Cellular Pathology, vol. 28, no. 5-6, pp. 283-294, 2006

Authors: Janssen, Emiel A.M. | van Diest, Paul J. | Søiland, Håvard | Gudlaugson, Einar | Nysted, Arne | Voorhorst, Feja J. | Vermorken, Jan B. | Søreide, Jon-Arne | Baak, Jan P.A.

Article Type: Research Article

Abstract: Background: Adjuvant systemic chemotherapy (ASCT) in lymph node-negative breast (LN−) cancers improves survival. The majority of (LN−) patients receive ASCT when the St. Gallen criteria or its modifications are used, as accurate identifiers which patients benefit from ASCT are lacking. This may imply over-treatment in many patients. Aim: To evaluate which patients or primary tumor factors predict ASCT success. Material and method: Retrospective analysis by single and multivariate survival analysis of clinical and tumor characteristics in (LN−) breast cancers <55 years, related to ASCT (n=125) or-not (n=516). Results: The two patient groups did not differ in age, tumor diameter, grade, …type, number of mitoses and other factors. Fourteen-year survival for the ASCT and non-ASCT patients was 83% and 74% (Hazard Ratio = HR = 0.33; p<0.0001, 9% absolute = 12% relative difference). Subgroup analysis showed that the recurrence-free survival = RFS of ASCT treated vs. non-treated patients differed in patients with grade 1 cancers (p=0.008), grade 2 cancers (p=0.004), grades 3 (p=0.02), tumors under and ≥2 cm (p=0.001 and 0.0002), oestrogen receptor-positive or -negative tumors (p=0.003,0.04), MAI < 10 and ≥10 (p=0.005,0.003) and fibrotic focus absent (p=0.002). With multivariate analysis the most important predictor of ASCT effect was the MAI. In patients with slowly proliferating tumors (MAI <3) no advantage was found between patients treated-or-not with adjuvant chemotherapy (RFS = 92% and 91%, p=0.13, p=0.63 for overall survival), contrasting those with MAI ≥3 (p=0.0001; HR = 0.32, 95% CI 0.18–0.58). Conclusion: MAI is the strongest predictor of adjuvant systemic chemotherapy success. In patients with MAI < 3 (31% of all patients), ASCT does not improve survival. Show more

Keywords: Breast cancer, proliferation, Mitotic Activity Index, prognosis, therapy

Citation: Analytical Cellular Pathology, vol. 28, no. 5-6, pp. 295-303, 2006

Authors: Henderson, Gregory S. | van Diest, Paul J. | Burger, Horst | Russo, Jose | Raman, Venu

Article Type: Research Article

Abstract: Introduction: Homeotic (HOX) gene products are now known to be functionally associated with breast cancer biogenesis. Recent evidence has indicated that HOXA5 regulates both p53 and progesterone receptor expression levels in breast cancer cells. In addition, HOXA5 has been shown to interact and regulate the activity of another protein referred to as Twist. As homeotic genes play a pivotal role in development, we sought to decipher the expression pattern in both normal breast tissues and in breast carcinomas. Methods: RT-PCR and immunohistochemistry were performed, to assay the levels of HOXA5 expression, on a panel of normal breast tissue and its …corresponding primary breast tumors. Results and Conclusions: We show that HOXA5 expression was maintained at stable levels at different reproductive stages of a woman's life, except during lactation. This evidence indicates that HOXA5 may play a role in maintaining the differentiated state within the breast epithelium. However, nearly 70% of all breast carcinomas had decreased HOXA5 protein levels as compared to normal breast tissues. In addition, we demonstrate that HOXA5 protein expression levels in breast carcinomas inversely co-relates with Epidermal Growth Factor Receptor (EGFR) expression. Furthermore, we found that the survival rate amongst the different low levels of HOXA5 expressing breast tumors was not significant, indicative of an early tumorigenesis process in the absence of innate levels of HOXA5 in normal breast cells. Show more

Keywords: Breast, homeotic gene, epidermal growth factor receptor

Citation: Analytical Cellular Pathology, vol. 28, no. 5-6, pp. 305-313, 2006

Authors: Skotheim, Rolf I. | Autio, Reija | Lind, Guro E. | Kraggerud, Sigrid M. | Andrews, Peter W. | Monni, Outi | Kallioniemi, Olli | Lothe, Ragnhild A.

Article Type: Research Article

Abstract: Introduction: Testicular germ cell tumors of adolescent and young adult men (TGCTs) generally have near triploid and complex karyotypes. The actual genes driving the tumorigenesis remain essentially to be identified. Materials and Methods: To determine the detailed DNA copy number changes, and investigate their impact on gene expression levels, we performed an integrated microarray profiling of TGCT genomes and transcriptomes. We analyzed 17 TGCTs, three precursor lesions, and the embryonal carcinoma cell lines, NTERA2 and 2102Ep, by comparative genomic hybridization microarrays (array-CGH), and integrated the data with transcriptome profiles of the same samples. Results: The gain of chromosome arm 12p …was, as expected, the most common aberration, and we found CCND2, CD9, GAPD, GDF3, NANOG, and TEAD4 to be the therein most highly over-expressed genes. Additional frequent genomic aberrations revealed some shorter chromosomal segments, which are novel to TGCT, as well as known aberrations for which we here refined boundaries. These include gains from 7p15.2 and 21q22.2, and losses of 4p16.3 and 22q13.3. Integration of DNA copy number information to gene expression profiles identified that BRCC3, FOS, MLLT11, NES, and RAC1 may act as novel oncogenes in TGCT. Similarly, DDX26, ERCC5, FZD4, NME4, OPTN, and RB1 were both lost and under-expressed genes, and are thus putative TGCT suppressor genes. Conclusion: This first genome-wide integrated array-CGH and gene expression profiling of TGCT provides novel insights into the genome biology underlying testicular tumorigenesis. Show more

Keywords: Array-CGH, comparative genomic hybridization, gene expression, microarray, testicular germ cell tumor

Citation: Analytical Cellular Pathology, vol. 28, no. 5-6, pp. 315-326, 2006