Browsing by Subject "Tissue microarray"
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- PublicationOpen AccessAnti-apoptotic activity in deep pelvic endometriosis(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2014) Abdalla Ribeiro, Helizabet S.; Galvão, Maria Antonieta Longo; Aoki, Tsutomu; Aldrighi, José Mendes; Ribeiro, Paulo AyrozaSince endometriosis is a proliferative disease we evaluated the presence of anti-apoptotic factor (Bcl2) and pro-apoptotic factor (Bax) in deep pelvic endometriosis. A Cross-sectional observational study was performed at Santa Casa de Misericórdia de São Paulo, São Paulo, Brazil. Forty women aged 26 to 46 years with deep endometriosis were selected. They had not been clinically treated for at least 3 months prior to surgery and then underwent surgical laparoscopy to treat the disease. During the surgery, tissue was collected from the uterosacral ligaments and the rectosigmoid; an endometrial biopsy was also performed as a control. All interventions were performed by the same surgeon. The specimens were sent for pathological and immunohistochemical analyses; endometriosis was confirmed in all patients. After the immunohistochemical reaction a semi-quantitative evaluation of the staining intensity (relative optical density-ROD) was conducted, applying the digital densitometric analysis system. In the uterosacral ligaments 97.5% of the specimens were positive for Bcl2 whereas in the rectosigmoid 100% were positive. In the endometrium we observed that 87.5% were positive for Bcl2. BAX expression was null in the rectosigmoid and in the endometrium. In the uterosacral ligaments 2.5% of the specimens expressed BAX. The relative optical density of Bcl2 was higher in the rectosigmoid and in the uterosacral ligament when compared to the endometrium, 0.141±0.002; 0.129±0.001, respectively (p<0.01). We concluded that the anti-apoptotic factor Bcl-2 was expressed in all studied specimens, but in a higher staining intensity in the rectosigmoid and in the uterossacral ligaments in comparison to the endometrium. The pro-apoptotic factor Bax had virtually no expression in the studied tissues.
- PublicationOpen AccessFucosyltransferase 8 expression in breast cancer patients: A high throughput tissue microarray analysis(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Yue, Liling; Han, Cuicui; Li, Zubin; Li, Xin; Liu, Deshui; Liu, Shulin; Yu, HaitaoThe aim of this study was to compare the expression of fucosyltransferase 8 (FUT8) in breast cancer tissue and to investigate the relationship between this marker with tumor progression and its applicability to differential diagnosis. An immunohistochemical study was performed for FUT8 using the tissue microarray technique. In addition, the mRNA and protein levels of FUT8 in the tissue were also tested by real-time PCR and Western blot. There was a significant difference in cytoplasmic expression of FUT8 between breast cancer tissue and matched normal tissue (p<0.001). The percent of FUT8 staining in breast cancer tissues ranging from negative, weak positive, positive and strong positive were 2.7%, 40.2%, 54% and 3.2%, respectively. High FUT8 protein expression correlated with lymphatic metastasis (p=0.008) and with stage status (p=0.039). We detected that reduced FUT8 expression correlated with disease-free survival (p=0.02) and overall survival (p=0.04) of breast cancer patients. Expression of FUT8 can stratify breast cancer tissue and may be considered a prognostic marker for breast cancer patients.
- PublicationOpen AccessGene expression analysis in sections and tissue microarrays of archival tissues by mRNA in situ hybridization(Murcia : F. Hernández, 2005) Henke, R.T.; Maitra, A.; Palk, S.; Wellstein, A.Altered expression of genes in diseased tissues can prognosticate a distinct natural progression of the disease as well as predict sensitivity or resistance to particular therapies. Archival tissues from patients with a known medical history and treatments are an invaluable resource to validate the utility of candidate genes for prognosis and prediction of therapy outcomes. However, stored tissues with associated long-term follow-up information typically are formalin-fixed, paraffinembedded specimen and this can severely restrict the methods applicable for gene expression analysis. We report here on the utility of tissue microarrays (TMAs) that use valuable tissues sparingly and provide a platform for simultaneous analysis of gene expression in several hundred samples. In particular, we describe a stable method applicable to mRNA expression screening in such archival tissues. TMAs are constructed from sections of small drill cores, taken from tissue blocks of archival tissues and multiple samples can thus be arranged on a single microscope slide. We used mRNA in situ hybridization (ISH) on >500 full sections and >100 TMAs for >10 different cDNAs that yielded >10,000 data points. We provide detailed experimental protocols that can be implemented without major hurdles in a molecular pathology laboratory and discuss quantitative analysis and the advantages and limitations of ISH. We conclude that gene expression analysis in archival tissues by ISH is reliable and particularly useful when no protein detection methods are available for a candidate gene.
- PublicationOpen AccessHigh expression of USP18 is associated with the growth of colorectal carcinoma(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2021) Zhang, Lin; Zhang, Ningning; Li, Xin; Wu, Wanxin; Zhang, Yanping; Wang, JingyuAim. To investigate whether USP18 can be used as a predictive marker for the diagnosis and development of colorectal cancer. Methods. The Gene Expression Omnibus (GEO) Dataset and the Cancer Genome Atlas (TCGA) database were used to select differential proteins for the ubiquitinspecific peptidases (USPs). The extensive target prediction and network analysis methods were used to assess the association with the USP18 interacting proteins, as well as the statistical correlation between USP18 and the clinical pathology parameters. The effects of USP18 on the proliferation of colorectal cancer were examined using CCK8. The effects of USP18 on the migration of colorectal cancer were examined using wound healing assays. Immunohistochemistry (IHC) was performed on the tissue microarray. Results. The results showed that the expression of USP18 was related to age (P=0.014). The positive rates of the USP18 protein in T1, T2, T3, and T4 were 0.00%, 22.92%, 78.38%, and 95.35%, respectively (P<0.00). The positive rates of the USP18 protein in I, II, III, and IV were 47.43%, 83.12%, 66.67%, and 100.00%, respectively (P<0.00). The Western blot assay showed that the expression of USP18 in colorectal cancer tissues was significantly higher than that in matched paracancerous tissues (P<0.05). The CCK8 experiments suggested that USP18 promoted the migration of CRC cells. Wound healing assays suggested that USP18 promoted the proliferation of CRC cells. Conclusion. This study showed that USP18 can promote the proliferation of colorectal cancer cells and might be a potential biomarker for the diagnosis of CRC.
- PublicationOpen AccessModerate-to-strong expression of FGFR3 and TP53 alterations in a subpopulation of choroid plexus tumors(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Granberg, Kirsi J.; Raita, Annina; Lehtinen, Birgitta; Tiihonen, Aliisa M.; Kesseli, Juha; Annala, Matti; Rodriguez-Martinez, Alejandra; Nordfors, Kristiina; Zhang, Wei; Visakorpi, Tapio; Nykter, Matti; Haapasalo, Hannu K.Deregulation of fibroblast growth factor receptor (FGFR) signaling is tightly associated with numerous human malignancies, including cancer. Indeed, FGFR inhibitors are being tested as anti-tumor drugs in clinical trials. Among gliomas, FGFR3 fusions occur in IDH wild-type diffuse gliomas leading to high FGFR3 protein expression and both, FGFR3 and FGFR1, show elevated expression in aggressive ependymomas. The aim of this study was to uncover the expression of FGFR1 and FGFR3 proteins in choroid plexus tumors and to further characterize FGFR-related as well as other genetic alterations in FGFR3 expressing tumors. Expression levels of FGFR1 and FGFR3 were detected in 15 choroid plexus tumor tissues using immunohistochemistry of tissue microarrays and 6 samples were subjected to whole mount FGFR3 staining. Targeted sequencing was used for deeper molecular analysis of two FGFR3 positive cases. Moderate expression of FGFR1 or FGFR3 was evidenced in one third of the studied choroid plexus tumors. Targeted sequencing of a choroid plexus carcinoma and an atypical choroid plexus papilloma, both with moderate-to-strong FGFR3 expression, revealed lack of protein-altering mutations or fusions in FGFR1 or FGFR3, but TP53 was altered in both tumors. FGFR3 and FGFR1 proteins are expressed in a subpopulation of choroid plexus tumors. Further studies using larger cohorts of patients will allow identification of the clinicopathological implications of FGFR1 and FGFR3 expression in choroid plexus tumors.
- PublicationOpen AccessTissue microarray validation in cervical carcinoma studies. A methodological approach(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2025) Lovane, Lucília; Carrilho, Carla; Karlsson, ChristinaTissue microarrays (TMAs) are a cost-effective tool to study biomarkers in clinical research. Cervical cancer (CC) is one of the most prevalent in women worldwide, with the highest prevalence in low-middle-income countries due to a lack of organized screening. CC is associated with persistent high-risk human papillomavirus infection. Several biomarkers have been studied for diagnostic, therapeutic, and prognostic purposes. We aimed to evaluate and validate the effectiveness of TMA in CC compared to whole slide images (WSs). We selected and anonymized twenty cases of CC. P16, cytokeratin 5 (CK5), cytokeratin 7 (CK7), programmed death-ligand 1 (PD-L1), and CD8 expression were immunohistochemically investigated. All WS were scanned and 10 representative virtual TMA cores with 0.6 mm diameter per sample were selected. Ten random combinations of 1-5 cylinders per case were assessed for each biomarker. The agreement of scoring between TMA and WS was evaluated by kappa statistics. We found that three cores of 0.6 mm on TMA can accurately represent WS in our setting. The Kappa value between TMA and WS varied from 1 for p16 to 0.61 for PD-L1. Our study presents an approach to address TMA sampling that could be generalized to TMA-based research, regardless of the tissue and biomarkers of interest.