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Unlocking breast cancer in Brazilian public health system: Using tissue microarray for accurate immunohistochemical evaluation with limitations in subtyping

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WOMENS HEALTH 2025年 21卷 17455057241304654-17455057241304654页

作者： Ruppenthal, Rubia Denise Pilar, Emily Ferreira Salles dos Santos, Jordan Boeira Coelho, Rafael Correa Henriques, Carina Machado Costamilan Uchoa, Diego de Mendonca Graudenz, Marcia Silveira Univ Fed Rio Grande do Sul Fac Med Dept Pathol Porto Alegre RS Brazil Hosp Clin Porto Alegre Lab Pathol Porto Alegre RS Brazil Univ Fed Rio Grande do Sul Fac Med Grad Program Sci Gastroenterol & Hepatol Porto Alegre RS Brazil Hosp Clin Porto Alegre Lab Expt Pathol Porto Alegre RS Brazil Univ Fed Rio Grande do Sul Fac Med Grad Program Med Sci Porto Alegre RS Brazil

Background: Breast cancer (BC) is a significant burden on healthcare systems, especially in low- and middle-income countries where access to diagnosis and treatment is challenging. Objectives: The purpose of this study was to assess the diagnostic accuracy and cost using tissue microarray (TMA) instead of traditional immunohistochemical (IHC) evaluation for estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor 2 (HER2), and the proliferation marker Ki-67 and BC subtyping within the Brazilian public health system. Design: This is a retrospective cohort study comparing TMA slides with traditional whole-slide evaluation for IHC markers in 242 BC cases. Methods: We used formalin-fixed tissue blocks for TMA assembly. Clinical data and IHC scores for ER, PR, HER2, and Ki-67 were obtained from pathology reports. Cohen's kappa (k) was used to assess TMA performance. Results: BC samples were distributed in 10 TMAs and 968 cores were scored (242 BC cases x 4 markers). In 97% of these, TMA reached high quality to adequate IHC scoring with minimal technical issues. Inter-examiner agreement was almost perfect for all markers (ranging from 0.85 for HER2 to 0.91 for ER, p < 0.001). The intratumoral heterogeneity ranged from almost perfect agreement for ER and HER2 to moderate to substantial for PR and Ki-67. TMA offers substantial time and cost savings, with an approximately 11-fold reduction compared to traditional methods. The concordance between TMA and original reports was almost perfect, with 93% overall agreement (k = 0.81, p < 0.001). However, TMA performance varied between markers, with intratumoral heterogeneity significantly impacting discordant results, particularly for Ki-67 and HER2. This ultimately affected the accuracy of BC subtyping. TMA performed well in identifying luminal A and triple-negative cases, but misclassification was common for luminal B and HER2-positive cases. Conclusion: TMA offers accurate and lower-cost results in the i

关键词： breast cancer immunohistochemistry tissue microarray public health subtyping

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tissue microarray analysis of interleukin-20 expression

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CYTOKINE 2006年第1-2期35卷 44-52页

作者： Hsing, Chung-Hsi Ho, Chung-Liang Chang, Lih-Yun Lee, Yi-Lin Chuang, Shih-Sung Chang, Ming-Shi Natl Cheng Kung Univ Inst Biopharmaceut Sci Coll Med Tainan 70101 Taiwan Natl Cheng Kung Univ Dept Pathol Coll Med Tainan 70101 Taiwan Natl Cheng Kung Univ Inst Biochem & Mol Biol Coll Med Tainan 70428 Taiwan Chi Mei Med Ctr Dept Med Res Tainan Taiwan Chi Mei Med Ctr Dept Anesthesiol Tainan Taiwan Chi Mei Med Ctr Dept Pathol Tainan Taiwan Taipei Med Univ Dept Pathol Taipei Taiwan

Knowledge about the biological functions and clinical implications of interleukin (IL)-20, a recently discovered cytokine in the IL-10 family, is still incomplete. Our aim was to determine the distribution of IL-20 expression and to delineate the cell types that express IL-20 in healthy and neoplastic tissue, because this information will significantly affect the exploration of its pathophysiological roles. We used tissue microarray technology and an immunohistochemical survey using an anti-IL-20 monoclonal antibody to examine IL-20 expression in 36 non-neoplastic and 14 neoplastic tissues. IL-20 protein was positively stained in 30 non-neoplastic tissue types and five major cell types: epithelial cells, myoepithelial cells, endothelial cells, macrophages, and skeletal muscle cells. We also found that several types of tumor cells stained positive for IL-20, especially in squamous cell carcinoma of the skin, tongue, esophagus, and lung. Our data provide valuable references for further investigation of the biological functions and clinical implications of IL-20 in humans. (c) 2006 Elsevier Ltd. All rights reserved.

关键词： cytokines interleukin-20 tissue microarray

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tissue microarray based analysis of prognostic markers in invasive bladder cancer:: Much effort to no avail

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UROLOGIC ONCOLOGY-SEMINARS AND ORIGINAL INVESTIGATIONS 2008年第1期26卷 17-24页

作者： Liedberg, Fredrik Anderson, Harald Chebil, Gunilla Gudjonsson, Sigurdur Hoglund, Mattias Lindgren, David Lundberg, Lena-Maria Lovgren, Kristina Ferno, Marten Mansson, Wiking Univ Lund Hosp Dept Urol S-22185 Lund Sweden Univ Lund Hosp Dept Canc Epidemiol S-22185 Lund Sweden Helsingborg Cty Hosp Dept Pathol Helsingborg Sweden Univ Lund Hosp Dept Clin Genet S-22185 Lund Sweden Univ Lund Hosp Dept Pathol S-22185 Lund Sweden Univ Lund Hosp Dept Oncol Med Sci S-22185 Lund Sweden

Purpose: To evaluate altered protein expression with tissue microarray methodology for 15 different markers with potential prognostic significance in invasive bladder cancer. Materials and Methods: Invasive tumor was sampled with the tissue-arraying instrument in 133 consecutive patients who underwent radical cystectomy, and at least 3, 0.6-min tissue cores were obtained. With immunohistochemistry, the expressions of TP53, RB1, CDKN1A (p21), MKI67 (Ki67), PTGS2 (Cox-2), CTNNA1 (alpha-catenin), CTNNB1 (beta-catenin), AKT, PTEN, RHOA, RHOC, STAT1, VEGFC, EGFR, and ERBB2 (HER2) were quantified, and correlations were made with tumor grade, pathologic stage, lymph node status, and disease-specific survival. Results: Decreased immunohistochemical expression of CTNNA1 and of PTEN correlated with higher pathologic tumor stages (P = 0.01 and P = 0.01, respectively), whereas increased AKT1 and ERBB2 correlated with lower pathologic tumor stages (P = 0.01 and P = 0.03, respectively). Increased RHOA expression was more common in grade 3 than in grade 2 tumors (P = 0.016). There were no other correlations among the 15 factors studied and pathologic stage, lymph node status, or tumor grade. No association was found between bladder cancer death and altered marker status for my of the markers studied. Conclusions: Currently, there are reasons to have a skeptical attitude toward the value of tissue microarray based immunohistochemistry as a method for evaluating prognostic markers in invasive bladder cancer. In this study, 15 antibodies were tested but were found to be of little clinical value. Whether this negative finding is related to the group of patients or factors studied, or the methodology is unclear. (c) 2008 Elsevier Inc. All rights reserved.

关键词： bladder cancer radical cystectomy prognosis tissue microarray

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tissue microarray and digital image analysis: a methodological study with special reference to the microenvironment in Hodgkin lymphoma

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HISTOPATHOLOGY 2012年第1期61卷 26-32页

作者： Glimelius, Ingrid Qvarnstrom, Fredrik Simonsson, Martin Ekwall, Anders Smedby, Karin E. Molin, Daniel Amini, Rose-Marie Uppsala Univ Dept Radiol Oncol & Radiat Sci Rudbeck Lab Uppsala Sweden Karolinska Inst Dept Med Solna Clin Epidemiol Unit Stockholm Sweden Uppsala Univ Dept Immunol Genet & Pathol Rudbeck Lab Uppsala Sweden

Glimelius I, Qvarnstrom F, Simonsson M, Ekwall A, Smedby K E, Molin D & Amini R-M ?(2012) Histopathology similar to 61, 2632 tissue microarray and digital image analysis: a methodological study with special reference to the microenvironment in Hodgkin lymphoma Aim: Cancer research has moved from solely investigating the tumour cells to also including analysis of the tumour microenvironment;however, the methods utilized have not been evaluated for this change. The aim of this study was to compare tissue microarrays (TMA) to whole tissue sections (WS) with regard to cells in the tumour microenvironment. Manual evaluation and digital image analyses (DIA) were utilized and also compared. Methods and results: TMA slides from 117 Hodgkin lymphoma patients were immunostained for forkhead box protein 3 (FoxP3) [identifying regulatory T cells (Treg)], and 39 corresponding WS were also analysed. Manual evaluation and DIA were utilized for all patients on both the TMA and the WS. A correlation coefficient of 0.83 was obtained for the proportion of Treg in TMA versus WS using manual evaluation and a correlation coefficient of 0.77 with DIA. Treg counts using manual evaluation correlated in turn with DIA, with a coefficient of 0.79 for the 117 TMA sections and 0.65 for the 39 WS. Conclusion: Because a high correlation was observed between TMA and WS, TMA can be utilized when evaluating cells in the tumour microenvironment. DIA appears to provide a reliable measurement method, provided that manual control of the tumour slides is conducted.

关键词： digital imaging Hodgkin lymphoma microenvironment regulatory T cells tissue microarray

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tissue microarray profiling in human heart failure

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PROTEOMICS 2016年第17期16卷 2319-2326页

作者： Lal, Sean Nguyen, Lisa Tezone, Rhenan Ponten, Fredrik Odeberg, Jacob Li, Amy dos Remedios, Cristobal Univ Sydney Bosch Inst Sydney Med Sch Dept Anat & Histol Sydney NSW Australia KTH Sch Biotechnol Royal Inst Technol Dept ProteSci Life Lab Stockholm Sweden Uppsala Univ Dept Immunol Genet & Pathol Sci Life Lab Uppsala Sweden

tissue microarrays (TMAs) are a versatile tool for high-throughput protein screening, allowing qualitative analysis of a large number of samples on a single slide. We have developed a customizable TMA system that uniquely utilizes cryopreserved human cardiac samples from both heart failure and donor patients to produce formalin-fixed paraffin-embedded sections. Confirmatory upstream or downstream molecular studies can then be performed on the same (biobanked) cryopreserved tissue. In a pilot study, we applied our TMAs to screen for the expression of four-and-a-half LIM-domain 2 (FHL2), a member of the four-and-a-half LIM family. This protein has been implicated in the pathogenesis of heart failure in a variety of animal models. While FHL2 is abundant in the heart, not much is known about its expression in human heart failure. For this purpose, we generated an affinity-purified rabbit polyclonal anti-human FHL2 antibody. Our TMAs allowed high-throughput profiling of FHL2 protein using qualitative and semiquantitative immunohistochemistry that proved complementary to Western blot analysis. We demonstrated a significant relative reduction in FHL2 protein expression across different forms of human heart failure.

关键词： Cardiomyopathy Human heart failure Lim proteins Protein arrays Proteomic profiling tissue microarray

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tissue microarray analysis of human FRATI expression and its correlation with the subcellular localisation of β-catenin in ovarian tumours

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BRITISH JOURNAL OF CANCER 2006年第5期94卷 686-691页

作者： Wang, Y Hewitt, SM Liu, S Zhou, X Zhu, H Zhou, C Zhang, G Quan, L Bai, J Xu, N Chinese Acad Med Sci Lab Cell & Mol Biol Inst Canc Beijing 100021 Peoples R China Chinese Acad Med Sci Lab Cell & Mol Biol Canc Hosp Beijing 100021 Peoples R China Peking Union Med Coll Beijing 100021 Peoples R China NCI NIH Tissue Array Res Program Pathol LabCtr Canc Res Bethesda MD 20892 USA

The mechanisms involved in the pathogenesis of ovarian cancer are poorly understood, but evidence suggests that aberrant activation of Wnt/ beta- catenin signalling pathway plays a significant role in this malignancy. However, the molecular defects that contribute to the activation of this pathway have not been elucidated. Frequently rearranged in advanced T- cell lymphomas- 1 ( FRAT1) is a candidate for the regulation of cytoplasmic beta- catenin. In this study, we developed in situ hybridisation probes to evaluate the presence of FRAT1 and used an anti- beta- catenin antibody to evaluate by immunohistochemistry the expression levels and subcellular localisation of b- catenin in ovarian cancer tissue microarrays. Expression of FRAT1 was found in some human normal tissues and 47% of ovarian adenocarcinomas. A total of 46% of ovarian serous adenocarcinomas were positive for FRAT1 expression. Accumulation of beta- catenin in the nucleus and/ or cytoplasm was observed in 55% ovarian adenocarcinomas and in 59% of serous adenocarcinomas. A significant association was observed in ovarian serous adenocarcinomas between FRAT1 and beta- catenin expression ( P < 0.01). These findings support that Wnt/ beta- catenin signalling may be aberrantly activated through FRAT1 overexpression in ovarian serous adenocarcinomas. The mechanism behind the overexpression of FRAT1 in ovarian serous adenocarcinomas and its significance is yet to be investigated.

关键词： FRATI beta-catenin Wnt pathway ovarian cancer tissue microarray

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tissue microarray technique is applicable to bone marrow biopsies of myeloproliferative neoplasms

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HISTOCHEMISTRY AND CELL BIOLOGY 2017年第1期147卷 75-82页

作者： Limberger, Kathrin A. Bogatyreva, Lioudmila Todorova, Rumyana Herde, Bettina Hauschke, Dieter Pahl, Heike L. Werner, Martin Aumann, Konrad Univ Freiburg Inst Surg Pathol Med Ctr Fac Med Breisacher Str 115a D-79106 Freiburg Germany Schwarzwaldbaar Klinikum Villingen Gen & Visceral Surg Villingen Schwenningen Germany Univ Freiburg Inst Med Biometry & Med Informat Med Ctr Fac Med Freiburg Germany Univ Freiburg Comprehens Canc Ctr Freiburg Fac Med Freiburg Germany Univ Freiburg Dept Med Hematol Oncol & Stem Cell Transplantat 1 Div Mol Hematol Med CtrFac Med Freiburg Germany

tissue microarray (TMA) technique is an established high-throughput method to analyze multiple tissue specimens in parallel. However, in order to obtain reliable results from immunohistochemical analyses of TMA blocks, cell composition of TMA spots must correspond to whole tissue sections (WTS) particularly in tissues with a heterogeneous cell composition as it is the case in myeloproliferative neoplasms (MPN). The aim of this study was to validate TMA of bone marrow biopsies from MPN patients. TMAs of MPN bone marrow biopsies (ET: n = 26, PV: n = 26, and PMF: n = 29) were compiled in triplicates and MPN-specific histological parameters were assessed. Results of TMA spots were compared with WTS' results using the intra-class correlation coefficient (ICC). Immunohistochemical NFE2 and calreticulin stainings of the TMA with quantitative evaluation were performed. TMA construction was technically successful with a loss of 10 % of all spots. ICC calculation revealed high to moderate correlations of TMA with WTS, especially the parameters that are typically affected in MPN tissue, e.g. cellularity of hematopoiesis (ICC 0.62-0.89), number of megakaryocytes (ICC 0.50-0.71), micro-vessel density (ICC 0.56-0.91), or grade of myelofibrosis (ICC 0.56-0.89). Results of NFE2 and calreticulin immunohistochemistry of MPN TMAs are consistent with previously published data. Overall, our results show moderate to good correlation between histological data of WTS and TMA spots illustrating that the TMA technique is applicable to bone marrow biopsies of MPN patients. However, TMA construction in triplicates is necessary to reach sufficient correlation. MPN TMAs can be applied for serial immunohistochemical surveys of archived tissues to assess the mutation status or to further sub-classify MPN cases.

关键词： tissue microarray Myeloproliferative neoplasms Validation Immunohistochemistry

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tissue microarray EVIDENCE OF ASSOCIATION BETWEEN p16 AND PHOSPHORYLATED eIF4E IN TONSILLAR SQUAMOUS CELL CARCINOMA

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HEAD AND NECK-JOURNAL FOR THE SCIENCES AND SPECIALTIES OF THE HEAD AND NECK 2011年第9期33卷 1340-1345页

作者： Fury, Matthew G. Drobnjak, Marija Sima, Camelia S. Asher, Marina Shah, Jatin Lee, Nancy Carlson, Diane Wendel, H. Guido Pfister, David G. Mem Sloan Kettering Canc Ctr Head & Neck Med Oncol Serv New York NY 10021 USA Mem Sloan Kettering Canc Ctr Dept Pathol New York NY 10021 USA Mem Sloan Kettering Canc Ctr Dept Epidemiol & Biostat New York NY 10021 USA Mem Sloan Kettering Canc Ctr Dept Surg New York NY 10021 USA Mem Sloan Kettering Canc Ctr Dept Radiat Oncol New York NY 10021 USA Mem Sloan Kettering Canc Ctr Canc Biol & Genet Program New York NY 10021 USA

Background. Expression of p16 is a marker for human papillomavirus (HPV)-related carcinogenesis in head and neck cancer. The purpose of this study is to determine if p16 immunoreactivity is associated with aberrant expression of components of the PI3 kinase pathway. Methods. A tissue microarray (TMA) was constructed for 46 archived tonsillar squamous cell carcinoma specimens. Clinical demographics of these patients were analyzed, and the TMA was interrogated with antibodies directed against p16, phosphorylated Akt(Ser473), phosphorylated S6(Ser240/244), phosphorylated S6(Ser235/236), phosphorylated 4E-BP1(Thr37/46), phosphorylated eIF4E(Ser209), PTEN, p21, and p53. Results. There was a significant correlation between history of tobacco abuse (> 10 pack/years) and absence of p16 expression (p = .01). Expression of p16 was significantly associated with immunoreactivity of p21 (p = .02), PTEN (p = .02), and phosphorylated eIF4E (p = .03). There was no evidence of association between p16 status and expression of phosphorylated S6, phosphorylated 4E-BP1, or p53. Conclusion. p16 positive tonsillar squamous cell carcinoma is characterized by expression of phosphorylated eIF4E that may occur via a mammalian target of rapamycin (mTOR)-independent mechanism. (C) 2010 Wiley Periodicals, Inc. Head Neck 33: 1340-1345, 2011

关键词： HPV tonsil p16 tissue microarray mTOR

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tissue microarray-based study of hepatocellular carcinoma validating SPIB as potential clinical prognostic marker

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ACTA HISTOCHEMICA 2016年第1期118卷 38-45页

作者： Ho, Yi-Jung Lin, Yueh-Min Huang, Yen-Chi Yeh, Kun-Tu Lin, Liang-In Lu, Jeng-Wei Natl Def Med Ctr Inst Prevent Med Taipei Taiwan Changhua Christian Hosp Dept Pathol Changhua Taiwan Jen Teh Jr Coll Med Nursing & Management Dept Med Technol Miaoli Taiwan Hsin Sheng Jr Coll Med Care & Management Dept Styling & Cosmetol Taoyuan Taiwan Chung Shan Med Univ Sch Med Taichung Taiwan Natl Taiwan Univ Hosp Dept Lab Med Taipei Taiwan Natl Taiwan Univ Dept Clin Lab Sci & Med Biotechnol Taipei 10764 Taiwan Natl Taiwan Univ Coll Med Dept Clin Lab Sci & Med Biotechnol Taipei Taiwan

Currently, the prognostic significance of SPIB protein overexpression in human hepatocellular carcinoma (HCC) is unclear. The aim of the present study was to investigate the level of SPIB expression in human HCC in order to determine possible correlations between SPIB expression and clinicopathological findings. The expression of SPIB proteins was detected using immunohistochemical staining in commercial multiple-tissue microarrays as a means of examining expression profiles in patients. Using online biomarker validation tool SurvExpress, we focused on the correlation between SPIB overexpression and survival as well as relapse-free survival (RFS). Results show that SPIB protein expression levels were significantly higher in colon, liver, and stomach tumors than in non-tumor tissues (p < 0.05). SPIB over expression in patients with HCC was also significantly higher than that of the normal samples (p < 0.001). Among patients with liver disease, SPIB protein expression levels differ significantly according to the stage of liver disease, specifically between stages I, II, and III of HCC (p < 0.05). SPIB expression was also shown to be significantly correlated with age (p = 0.046) and histological grade (p = 0.027). Furthermore, the SurvExpress analysis suggested that high SPIB and KI-67 mRNA expression were significantly associated with the poor survival of patients with HCC (p < 0.05). Our results indicate that cross-talk in the expression of SPIB and KI-67 may be associated with poor prognosis and may potentially serve as a clinical prognostic indicator of HCC. This is the first time that such an association has been reported. (C) 2015 Elsevier GmbH. All rights reserved.

关键词： Prognostic marker Immunohistochemistry SPIB tissue microarray Hepatocellular carcinoma

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tissue microarray of head and neck squamous carcinoma: validation of the methodology for the study of cutaneous fatty acid-binding protein, vascular endothelial growth factor, involucrin and Ki-67

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VIRCHOWS ARCHIV 2005年第4期447卷 701-709页

作者： Gomaa, W Ke, YQ Fujii, H Helliwell, T Univ Liverpool Dept Pathol Liverpool L69 3GA Merseyside England Niigata Univ Sch Med Dept Biochem Niigata 951 Japan

tissue microarrays allow the simultaneous analysis of many tumours using small-diameter cores sampled from larger blocks of tissue, but may be limited by tumour heterogeneity. This study considers the validation of tissue microarray for the study of four molecules of interest as prognostic factors in head and neck squamous carcinoma, including a consideration of methods for assessing immunocytochemical scoring of microarrays. tissue microarray blocks were constructed from 100 cases of head and neck squamous carcinoma, taking four cores from different areas of each tumour. Immunocytochemical labelling was performed for cutaneous fatty acid binding protein, involucrin, vascular endothelial growth factor and Ki-67. The extent and intensity of scoring was determined for each core and the degree of agreement determined for results from the assessment of two, three or four cores for each carcinoma. In a subset of 30 representative cases, the labelling in the tissue microarrays was compared with that in whole-tissue sections of the same carcinomas. An adequate sample of carcinoma was achieved in more than 90% of the 400 cores;unsuccessful results were attributed to uneven core alignment or to poor targeting of the tumour tissue in the donor blocks. The degree of agreement in the assessment of extent and intensity of labelling was moderate to good (weighted kappa, range 0.479-0.902) between whole-tissue sections and microarray sections depending on the antigen and the scoring system. tissue microarray is a reliable tool to demonstrate cellular and molecular alterations in head and neck squamous carcinomas. We recommend using the mean results from four cores for biological studies, with analysis of categorical data based on quartile groups. Concordance with whole-tissue section data is reassuring, but data from microarrays need to be validated against clinical outcomes.

关键词： head and neck squamous carcinoma tissue microarray validation immunocytochemistry cutaneous fatty acid binding protein proliferation

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