Analytical Cellular Pathology - Volume 33, issue 1

Authors: Fariña-Sarasqueta, A. | Gosens, M.J.E.M. | Moerland, E. | van Lijnschoten, I. | Lemmens, V.E.P.P. | Slooter, G.D. | Rutten, H.J.T. | van den Brule, A.J.C.

Article Type: Research Article

Abstract: Aim: Although the predictive and prognostic value of thymidylate synthase (TS) expression and gene polymorphism in colon cancer has been widely studied, the results are inconclusive probably because of methodological differences. With this study, we aimed to elucidate the role of TS gene polymorphisms genotyping in therapy response in stage III colon carcinoma patients treated with 5-FU adjuvant chemotherapy. Patients and methods: 251 patients diagnosed with stage III colon carcinoma treated with surgery followed by 5-FU based adjuvant therapy were selected. The variable number of tandem repeats (VNTR) and the single nucleotide polymorphism (SNP) in the 5′-untranslated region of …the TS gene were genotyped. Results: There was a positive association between tumor T stage and the VNTR genotypes (p=0.05). In both univariate and multivariate survival analysis no effects of the studied polymorphisms on survival were found. However, there was an association between both polymorphisms and age. Among patients younger than 60 years, the patients homozygous for 2R seemed to have a better overall survival, whereas among the patients older than 67 this longer survival was seen by the carriers of other genotypes. Conclusion: We conclude that the TS VNTR and SNP do not predict response to 5-FU therapy in patients with stage III colon carcinoma. However, age appears to modify the effects of TS polymorphisms on survival. Show more

Keywords: Colon carcinoma, TS, VNTR, SNP, survival, 5-FU, age

DOI: 10.3233/ACP-CLO-2010-0526

Citation: Analytical Cellular Pathology, vol. 33, no. 1, pp. 1-11, 2010

Authors: Moelans, Cathy B. | Monsuur, Hanneke N. | de Pinth, Johannes H. | Radersma, Remco D. | de Weger, Roel A. | van Diest, Paul J.

Article Type: Research Article

Abstract: Background: Expression of estrogen receptor alpha (ERα) is predictive for endocrine therapy response and an important prognostic factor in breast cancer. Overexpression of ERα can be caused by estrogen receptor 1 (ESR1) gene amplification and was originally reported to be a frequent event associated with a significantly longer survival for ER-positive women treated with adjuvant tamoxifen monotherapy, which was however questioned by subsequent studies. Methods: This study aimed to reanalyze the frequency of ESR1 amplification by multiplex ligation-dependent probe amplification (MLPA) and fluorescence in situ hybridisation (FISH), and to assess clinicopathologic correlations. MLPA was performed in a group of …135 breast cancer patients, and gains/amplifications were subjected to FISH. Results: True ESR1 amplification by MLPA was rare (2%) and only 6% more patients showed a modest gain of ESR1. All MLPA-detected ESR1 amplifications and nearly all ESR1 gains were also FISH amplified and gained, but not all FISH amplifications/gains were MLPA amplified/gained, leading to an overall concordance of only 60% between both techniques. All 3 MLPA and FISH ESR1 amplified cases had high ERα expression, but there was no obvious correlation between ESR1 gain and ER status by IHC. ESR1 gains/amplifications were not associated with HER2 gain/amplification, but seemed to be associated with older age. Surprisingly, ESR1 gain/amplification was not associated with low grade as reported previously, but correlated with high grade and high proliferation. Furthermore, ESR1 gain/amplification by MLPA was not associated with nodal status or tumor size (pT status). Conclusions: ESR1 amplification as detected by MLPA is rare in breast cancer, and seems to be associated with high ERα expression, high age, high grade and high proliferation. This study confirms previous studies that showed differences in the ESR1 amplification frequencies detected by different techniques. Show more

Keywords: ESR1, MLPA, breast cancer, amplification

DOI: 10.3233/ACP-CLO-2010-0527

Citation: Analytical Cellular Pathology, vol. 33, no. 1, pp. 13-18, 2010

Authors: Chatzistamou, Ioulia | Dioufa, Nikolina | Trimis, George | Sklavounou, Alexandra | Kittas, Christos | Kiaris, Hippokratis | Papavassiliou, Athanasios G.

Article Type: Research Article

Abstract: Background: Concerted alterations between stromal fibroblasts and neoplastic cells underline the carcinogenic process. Activation of alpha-smooth muscle actin (SMA) expression, a cytoskeleton protein normally expressed only in myoepithelial cells, is considered a landmark for the activation of stromal fibroblasts with little however being known regarding the mechanism governing the expression of SMA in the stroma. Methods: We have evaluated by immunohistochemistry the expression of SMA in the stroma of oral malignant and pre-malignant lesions, in association with the expression of p53 and p21 tumor suppressors that were shown previously to be deregulated and/or mutated in stromal fibroblasts of various …cancers. The effects of p21 knockdown in SMA expression and cell migration and the mRNA levels of endogenous p21 in fibroblasts co-cultured with cancer cells were also assessed. Results: We found that both p21 and SMA expression was elevated in the stroma, but not the epithelium, of malignant as compared to pre-malignant lesions. We also noted that the expression of both was positively correlated, implying that SMA expression may be regulated by p21. Consistently with this notion we found that siRNA-mediated p21 suppression resulted in the reduction of SMA levels and also inhibited cell migration. Conclusion: Our results show that p21 deregulation is associated with the activation of stromal fibroblasts of oral cancers by a mechanism that involves the stimulation of SMA expression. Show more

Keywords: Stroma, p21, SMA, fibroblasts, desmoplastic reaction

DOI: 10.3233/ACP-CLO-2010-0528

Citation: Analytical Cellular Pathology, vol. 33, no. 1, pp. 19-26, 2010

Authors: Mazzucchelli, Roberta | Morichetti, Doriana | Santinelli, Alfredo | Scarpelli, Marina | Bono, Aldo V. | Lopez-Beltran, Antonio | Cheng, Liang | Montironi, Rodolfo

Article Type: Research Article

Abstract: Objective: The aim was to examine the expression and localization of the five somatostatin receptors (termed SSTR1–5) in radical prostatectomies (RPs) from patients with prostatic adenocarcinoma (PCa) under complete androgen ablation (CAA) before operation. Material: The five SSTRs were evaluated in the epithelial, smooth muscle and endothelial cells of normal-looking epithelium (Nep), high-grade prostatic intraepithelial neoplasia (HGPIN) and PCa in 20 RPs with clinically detected PCa from patients under CAA. 20 RPs with clinically detected PCa from hormonally untreated patients were used as control group. Results: Concerning the secretory cells (i) membrane staining was seen for SSTR3 and …SSTR4; the mean percentages of positive cells, higher in SSTR3 than in SSTR4, decreased sharply in HGPIN and PCa compared with Nep; the mean percentages in the androgen ablated group were 30–90% lower than in the untreated; (ii) cytoplasmic staining was seen for all 5 SSTRs; the mean percentages of positive cells in Nep, HGPIN and PCa of the untreated group were similar, and in general as high as 80% or more; in the treated group, the Nep values were similar to those in the untreated, whereas the values in HGPIN and PCa were lower for SSTR1, 3 and 5, with a decrease of 30% for SSTR1; (iii) nuclear staining was seen with SSTR4 and SSTR5, the mean percentages for the former being much lower than for the latter; treatment affected both HGPIN and PCa, whose proportions of stained cells were 30–55% lower than in the untreated group. Cytoplasmic staining in the basal cells was seen for all 5 SSTRs, both in Nep and HGPIN. The values in the treated group were lower than in the other, the difference between the two group being in general comprised between 10 and 40%. Treatment did not affect SSTR staining in the smooth muscle and endothelial cells. Conclusions: The present study expands our knowledge on the expression and localization of the five SSTRs in the prostate following CAA. Show more

Keywords: Somatostatin receptors, prostate cancer, high-grade prostatic intraepithelial neoplasia, complete androgen ablation

DOI: 10.3233/ACP-CLO-2010-0529

Citation: Analytical Cellular Pathology, vol. 33, no. 1, pp. 27-36, 2010

Authors: Klink, Barbara | Schlingelhof, Ben | Klink, Martin | Stout-Weider, Karen | Patt, Stephan | Schrock, Evelin

Article Type: Research Article

Abstract: Background: Glioblastomas are the most common and most malignant brain tumors in adults. A small subgroup of glioblastomas contains areas with histological features of oligodendroglial differentiation (GBMO). Our objective was to genetically characterize the oligodendroglial and the astrocytic parts of GBMOs and correlate morphologic and genetic features with clinical data. Methods: The oligodendroglial and the “classic” glioblastoma parts of 13 GBMO were analyzed separately by interphase fluorescence in situ hybridization (FISH) on paraffin sections using a custom probe set (regions 1p, 1q, 7q, 10q, 17p, 19q, cen18, 21q) and by comparative genomic hybridization (CGH) of microdissected paraffin embedded tumor …tissue. Results: We identified four distinct genetic subtypes in 13 GBMOs: an “astrocytic” subtype (9/13) characterized by +7/−10; an “oligodendroglial” subtype with −1p/−19q (1/13); an “intermediate” subtype showing +7/−1p (1/13), and an “other” subtype having none of the former aberrations typical for gliomas (2/13). The different histological tumor parts of GBMO revealed common genetic changes in all tumors and showed additional aberrations specific for each part. Conclusion: Our findings demonstrate the monoclonal origin of GBMO followed by the development of the astrocytic and oligodendroglial components. The diagnostic determination of the genetic signatures may allow for a better prognostication of the patients. Show more

Keywords: Glioblastoma, oligodendroglial component, GBMO, genetics, CGH, Interphase-FISH, genetic subclassification

DOI: 10.3233/ACP-CLO-2010-0530

Citation: Analytical Cellular Pathology, vol. 33, no. 1, pp. 37-54, 2010