scholarly journals FluorescenceIn SituHybridization for MicroRNA Detection in Archived Oral Cancer Tissues

2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
Zonggao Shi ◽  
Jeffrey J. Johnson ◽  
M. Sharon Stack
Keyword(s):  
Oral Cancer ◽  
Noncoding Rna ◽  
Diagnostic Value ◽  
Locked Nucleic Acid ◽  
Cancer Tissue ◽  
Dna Oligonucleotides ◽  
Cancer Pathology ◽  
Ffpe Tissues ◽  
Cancer Tissues

The noncoding RNA designated as microRNA (miRNA) is a large group of small single-stranded regulatory RNA and has generated wide-spread interest in human disease studies. To facilitate delineating the role of microRNAs in cancer pathology, we sought to explore the feasibility of detecting microRNA expression in formalin-fixed paraffin-embedded (FFPE) tissues. Using FFPE materials, we have compared fluorescentin situhybridization (FISH) procedures to detect miR-146a with (a) different synthetic probes: regular custom DNA oligonucleotides versus locked nucleic acid (LNA) incorporated DNA oligonucleotides; (b) different reporters for the probes: biotin versus digoxigenin (DIG); (c) different visualization: traditional versus tyramide signal amplification (TSA) system; (d) different blocking reagents for endogenous peroxidase. Finally, we performed miR-146a FISH on a commercially available oral cancer tissue microarray, which contains 40 cases of oral squamous cell carcinoma (OSCC) and 10 cases of normal epithelia from the human oral cavity. A sample FISH protocol for detecting miR-146a is provided. In summary, we have established reliablein situhybridization procedures for detecting the expression of microRNA in FFPE oral cancer tissues. This method is an important tool for studies on the involvement of microRNA in oral cancer pathology and may have potential prognostic or diagnostic value.

scholarly journals Overexpression of Keratin17 is Associated With Prognosis of Oral Cancer in the Chinese Population

2020 ◽  
Author(s):  
Lili Wang ◽  
Hongguang Song ◽  
Shiming Yang
Keyword(s):  
Oral Cancer ◽  
Prognostic Value ◽  
Cox Regression ◽  
Prognostic Biomarker ◽  
Human Cancer ◽  
Clinical Stage ◽  
Cancer Tissue ◽  
Cox Regression Analysis ◽  
Kaplan Meier ◽  
Cancer Tissues

Abstract Background: Oral cancer represents an common cancer among head and neck malignancies. The early metastasis contributes the poor prognosis of this disease. Keratin (KRT) family has ever been found to be involved in some crucial biological processes in normal cells and human cancer cells. As a member of this family, keratin 17 (KRT17) has been studied in some different cancers. In the present study, we aimed at assess the prognostic value of KRT17 in patients with oral cancer.Methods: Expression of KRT17 in 135 paired oral cancer tissues and noncancerous tissues was estimated by using quantitative Real-Time RT-PCR (qRT-PCR). The prognostic value of KRT17 was assessed with Kaplan-Meier survival analysis and multivariate Cox regression analysis.Results: Upregulated expression of KRT17 was detected in oral cancer tissues compared with the matched normal tissues (P<0.001). The expression of KRT17 was found associated with lymph node metastasis (P=0.025) and clinical stage (P=0.005). The Kaplan-Meier survival curves showed that the patients with high KRT17 expression possessed worse overall survival than those with low level cases (log-rank P=0.000). From the data of Cox analysis, we considered the KRT17 expression was an independent prognostic biomarker in oral cancer patients with HR of 2.489, 95% CI of 1.333-4.648 and P of 0.004.Conclusion: In summary, the expression of KRT17 was found upregulated in oral cancer tissue specimens and it was proved to serve as an independent prognostic biomarker in patients with oral cancer.

scholarly journals Overexpression of NNT-AS1 Activates TGF-β Signaling to Decrease Tumor CD4 Lymphocyte Infiltration in Hepatocellular Carcinoma

2020 ◽  
Vol 2020 ◽  
pp. 1-11
Author(s):  
Yakun Wang ◽  
Lei Yang ◽  
Xichen Dong ◽  
Xin Yang ◽  
Xinxue Zhang ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Signaling Pathway ◽  
Noncoding Rna ◽  
Tumor Infiltrating Lymphocytes ◽  
Cancer Tissue ◽  
Tumor Immune Evasion ◽  
Normal Tissues ◽  
Mechanistic Investigation ◽  
Cd4 Lymphocyte ◽  
Cancer Tissues

Nicotinamide nucleotide transhydrogenase-antisense RNA1 (NNT-AS1) is a long noncoding RNA (lncRNA) that has been shown to be overexpressed in hepatocellular carcinoma (HCC). However, the molecular mechanism involving NNT-AS1 in HCC remains to be extensively investigated. The activation of TGF-β signaling inhibits tumor-infiltrating lymphocytes (TILs) and results in tumor immune evasion. We thus planned to explore the mechanism by which NNT-AS1 activates the TGF-β signaling pathway and inhibits TILs in HCC. High levels of NNT-AS1 were detected in HCC tissues by both RNAscope and real-time quantitative PCR (RT-qPCR) assays. The levels of proteins involved in TGF-β signaling and those of CD4 T lymphocytes were quantified by immunohistochemistry (IHC). HCC cell lines (HepG2 and Huh7) were used to explore the effects of NNT-AS1 on TGF-β signaling activation. In these analyses, RNAscope detection demonstrated that NNT-AS1 levels were significantly increased in HCC cancer tissues ( P = 0.0001 ). In addition, the elevated NNT-AS1 levels in cancer tissue were further confirmed by RT-qPCR analysis of HCC cancer tissues ( n = 64 ) and normal tissues ( n = 26 ) ( P = 0.0003 ). Importantly, the overall survival time of HCC patients who exhibited higher levels of NNT-AS1 expression was significantly shorter than that of HCC patients who had lower levels of NNT-AS1 expression ( P = 0.0402 ). Further mechanistic investigation indicated that NNT-AS1 inhibition significantly decreased the levels of TGF-β, TGFBR1, and SMAD5 in HCC cells. In HCC tissues, IHC detection showed that relatively high NNT-AS1 levels were associated with a reduction in infiltrated CD4 lymphocyte numbers. In conclusion, this research identifies a novel mechanism by which NNT-AS1 impairs CD4 T cell infiltration via activation of the TGF-β signaling pathway in HCC.

scholarly journals Long Noncoding RNA NEAT1 as a Potential Candidate Biomarker for Prostate Cancer

Life ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 320
Author(s):  
Diana Nitusca ◽  
Anca Marcu ◽  
Alis Dema ◽  
Loredana Balacescu ◽  
Ovidiu Balacescu ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Noncoding Rna ◽  
Area Under The Curve ◽  
Diagnostic Value ◽  
Diagnostic Methods ◽  
Potential Candidate ◽  
Diagnostic Biomarkers ◽  
Potential Biomarker ◽  
Formalin Fixed Paraffin Embedded ◽  
Ffpe Tissues

Background: Prostate cancer (PCa) remains one of the leading causes of cancer-related mortality in men worldwide, mainly due to unsatisfactory diagnostic methods used at present, which lead to overdiagnosis, unnecessary biopsies and treatment, or misdiagnosis in early asymptomatic stages. New diagnostic biomarkers are needed for a correct and early diagnosis. Long noncoding RNAs (lncRNAs) have been broadly studied for their involvement in PCa biology, as well as for their potential role as diagnostic biomarkers. Methods: We conducted lncRNA profiling in plasma and microdissected formalin-fixed paraffin-embedded (FFPE) tissues of PCa patients and attempted validation for commonly dysregulated individual lncRNAs. Results: Plasma profiling revealed eight dysregulated lncRNAs, while microarray analysis revealed 717 significantly dysregulated lncRNAs, out of which only nuclear-enriched abundant transcript 1 (NEAT1) was commonly upregulated in plasma samples and FFPE tissues. NEAT1’s individual validation revealed statistically significant upregulation (FC = 2.101, p = 0.009). Receiver operating characteristic (ROC) analysis showed an area under the curve (AUC) value of 0.7298 for NEAT1 (95% CI = 0.5812–0.8785), suggesting a relatively high diagnostic value, thus having a potential biomarker role for this malignancy. Conclusions: We present herein data suggesting that NEAT1 could serve as a diagnostic biomarker for PCa. Additional studies of larger cohorts are needed to confirm our findings, as well as the oncogenic mechanism of NEAT1 in the development of PCa.

scholarly journals Study of Long Noncoding RNA FER1L4 and RB1, as Its Competing Endogenous RNA Network Target Gene, in Breast Cancer

2019 ◽  
Vol 12 (4) ◽  
pp. 21-30
Author(s):  
Zeinab Shaghaghi Torkdari ◽  
Mohammad Khalaj-Kondori ◽  
Mohammad Ali Hosseinpour Feizi
Keyword(s):  
Breast Cancer ◽  
Noncoding Rna ◽  
Target Gene ◽  
Ductal Carcinoma ◽  
Cancer Tissue ◽  
Breast Cancer Patients ◽  
Noncancerous Tissue ◽  
Tumor Tissues ◽  
Competing Endogenous Rna Network

Introduction: Breast cancer is the second most common cause of cancer-related death among females, which requires an exploration for markers to propose a more specific categorization of this cancer. Long noncoding RNAs (lncRNAs), the main subset of noncoding transcripts, are involved in tumorigenic processes. In this study, we investigated the expression of the fer-­1–­like family member 4 (FER1L4) lncRNA and its competitive endogenous RNA network target gene, RB transcriptional corepressor 1 (RB1), in ductal carcinoma (invasive and in situ) tissue and its adjacent noncancerous tissue (ANCT). Furthermore, associations of FER1L4 and RB1 with various clinical features of the patients were analyzed. Methods: Quantitative real-time PCR was used to measure the expression of the mentioned genes in 61 samples of ductal carcinoma and their ANCTs, and the data were analyzed using ANOVA and t tests. Results: FER1L expression was not significantly different in breast tumor samples compared with their ANCT samples, while RB1 showed significant downregulation in tumor tissues (P = 0.008). In addition, increased expression of FER1L4 and decreased RB1 expression were significantly correlated with lymph node metastasis in breast cancer patients (P < 0.05). Conclusion: FER1L4 is not upregulated in breast cancer tissue. However, RB1 expression is significantly downregulated.

scholarly journals Increased SPHK1 and HAS2 Expressions Correlate to Poor Prognosis in Pancreatic Cancer

2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Mengsi Yu ◽  
Kainan Zhang ◽  
Song Wang ◽  
Li Xue ◽  
Zhaoyun Chen ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
Cancer Patients ◽  
Cell Lines ◽  
Roc Curves ◽  
Diagnostic Value ◽  
Receptor Interaction ◽  
Cancer Tissue ◽  
Kaplan Meier ◽  
Short Overall Survival ◽  
Cancer Tissues

Objective. SPHK1 and HAS2 have been reported to play important roles in tumorigenesis and development. However, their expression and prognostic value in pancreatic cancer (PC) remain unclear. This study is aimed at investigating the expression of SPHK1 and HAS2 on the prognosis of pancreatic cancer. Materials and Methods. The expression of SPHK1 and HAS2 in pancreatic cancer tissues was analyzed through TCGA and GTEx databases and validated by qRT-PCR and Western blot in pancreatic cancer cell lines. χ 2 test was used to explore the correlation of the SPHK1 and HAS2 expressions with clinical characteristics. Kaplan-Meier survival analysis and ROC curve were used to evaluate the prognostic and diagnostic roles of SPHK1 and HAS2 in pancreatic cancer. Additionally, Spearman correlation analysis was applied to assess the correlation between the SPHK1 and HAS2 in pancreatic cancer. GO analysis and KEGG analysis were applied to explore the possible signaling pathway that SPHK1 and HAS2 coregulated genes mediated. Results. The expression of SPHK1 and HAS2 was markedly upregulated in pancreatic cancer tissue and cell lines, respectively. Furthermore, there was a significant positive correlation between SPHK1 and HAS2 expressions. ROC curves showed that SPHK1 combine with HAS2 has good diagnostic value in pancreatic cancer patients with 85% sensitivity and 99.4% specificity. Kaplan-Meier analysis showed that increased expression of SPHK1 and HAS2 was significantly associated with short overall survival (OS) of pancreatic cancer patients. GO and KEGG results revealed that SPHK1 and HAS2 mainly involved cell proliferation and invasion mediated by extracellular matrix- (ECM-) receptor interaction, focal adhesion, and PI3K-AKT signaling pathways. Conclusions. Overexpression of SPHK1 and HAS2 could be important markers for the prognosis of pancreatic cancer.

A PILOT STUDY OF HER-2/NEU GENE AMPLIFICATION ASSESSED BY FLUORESCENCE IN SITU HYBRIDIZATION (FISH) IN GASTRIC CANCER

2014 ◽  
pp. 15-20
Author(s):  
Van Huy Tran ◽  
Thi Minh Thi Ha ◽  
Trung Nghia Van ◽  
Viet Nhan Nguyen ◽  
Phan Tuong Quynh Le ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Cancer Patients ◽  
Gene Amplification ◽  
Cancer Tissue ◽  
Tissue Samples ◽  
Her 2 ◽  
Gastric Cancer Patients

Background: HER-2/neu is a predictive biomarker for treatment of gastric cancer using trastuzumab in combination with chemotherapy. This study aimed to evaluate the status of HER-2/neu gene amplification using fluorescence in situ hybridization (FISH) in gastric cancer. Patients and methods: thirty six gastric cancer patients were assessed HER-2/neu gene amplification by FISH using PathVysionTM HER-2 DNA Probe kit (including HER-2/neu probe and CEP-17 probe) with biopsy and surgical specimens. Results: The HER-2/neu gene amplification was observed in three cases (8.3%), the HER-2/neu gene amplification rate in Lauren’s intestinal-type and diffuse-type were 11.8% and 5.2%, respectively. Conclusion: We applied successfully FISH technique with gastric cancer tissue samples. This technique could be performed as routine test in gastric cancer in order to select patients that benefit from trastuzumab in combination with chemotherapy.

Aberrant β-catenin activity in hepatoid adenocarcinoma of the stomach

2020 ◽  
Vol 20 ◽  
Author(s):  
Nan Wang ◽  
Rui Kong ◽  
Wei Han ◽  
Jie Lu
Keyword(s):  
Hepatocellular Carcinoma ◽  
Pearson Correlation ◽  
Clinicopathological Characteristics ◽  
Cancer Tissue ◽  
Hepatoid Adenocarcinoma ◽  
Tumor Initiating Cells ◽  
Significant Difference ◽  
Diagnostic Confusion ◽  
Hematoxylin And Eosin ◽  
Cancer Tissues

Background: Hepatoid adenocarcinoma of the stomach (HAS) has been recognized as a rare primary gastric tumor characterized by hepatocellular carcinoma-like histology. HAS often causes diagnostic confusion with conventional gastric adenocarcinoma (CGA) due to the difficulty to detect hepatoid differentiation solely based on findings from hematoxylin and eosin (H&E) staining. Hence, HAS should be distinguished from solid-type CGA based on their different biological behaviors. β-catenin is highly expressed in hepatocellular carcinoma (HCC), which is involved in the maintenance of tumor initiating cells, drug resistance, tumor progression, and metastasis. Methods and Results: Given the dearth of HAS cases, systematic examination of the expression of β-catenin in HAS remains under-explored. In this study, 14 cases were subjected to immunostaining with with AFP, β-catenin, glypican3, hepar-1 and CerbB-2. In parallel, the clinicopathological characteristics of these patients were collected. We detected statistically significant difference in the expression of β-catenin (P = 0.000), glypican3 (P = 0.019), and hepar-1 (P = 0.007) between HAS cancer tissues and the adjacent non-cancerous tissues. Furthermore, a significant correlation was observed between the expression of β-catenin in HAS cancer tissue and adjacent tissue (Pearson correlation coefficient: 0.686, P = 0.007). Moreover, in cancer tissues, a significant correlation was observed between the expression of β-catenin and survival time (Spearman correlationcoefficient= - 0.482, P = 0.003). However, we found the expression of β-catenin did not correlate with the degree of tumor differentiation and tumor size, age, gender, serum AFP levels, microinvasion, and metastasis (P > 0.05). Conclusion: Our findings establish β-catenin as a useful marker that can distinguish HAS from CGA and may improve the early diagnosis to guide the appropriate and timely treatment of HAS patients.

scholarly journals New from Old: Thorectandrin Alkaloids in a Southern Australian Marine Sponge, Thorectandra choanoides (CMB-01889)

Marine Drugs ◽  
2021 ◽  
Vol 19 (2) ◽  
pp. 97
Author(s):  
Shamsunnahar Khushi ◽  
Angela A. Salim ◽  
Ahmed H. Elbanna ◽  
Laizuman Nahar ◽  
Robert J. Capon
Keyword(s):  
Marine Sponge ◽  
Tissue Specificity ◽  
Spectroscopic Analysis ◽  
Transformation Products ◽  
Chemical Fractionation ◽  
Cancer Tissue ◽  
Michael Acceptor ◽  
Michael Acceptors ◽  
Stable Ring

Thorectandra choanoides (CMB-01889) was prioritized as a source of promising new chemistry from a library of 960 southern Australian marine sponge extracts, using a global natural products social (GNPS) molecular networking approach. The sponge was collected at a depth of 45 m. Chemical fractionation followed by detailed spectroscopic analysis led to the discovery of a new tryptophan-derived alkaloid, thorectandrin A (1), with the GNPS cluster revealing a halo of related alkaloids 1a–1n. In considering biosynthetic origins, we propose that Thorectandrachoanoides (CMB-01889) produces four well-known alkaloids, 6-bromo-1′,8-dihydroaplysinopsin (2), 6-bromoaplysinopsin (3), aplysinopsin (4), and 1′,8-dihydroaplysinopsin (10), all of which are susceptible to processing by a putative indoleamine 2,3-dioxygenase-like (IDO) enzyme to 1a–1n. Where the 1′,8-dihydroalkaloids 2 and 10 are fully transformed to stable ring-opened thorectandrins 1 and 1a–1b, and 1h–1j, respectively, the conjugated precursors 3 and 4 are transformed to highly reactive Michael acceptors that during extraction and handling undergo complete transformation to artifacts 1c–1g, and 1k–1n, respectively. Knowledge of the susceptibility of aplysinopsins as substrates for IDOs, and the relative reactivity of Michael acceptor transformation products, informs our understanding of the pharmaceutical potential of this vintage marine pharmacophore. For example, the cancer tissue specificity of IDOs could be exploited for an immunotherapeutic response, with aplysinopsins transforming in situ to Michael acceptor thorectandrins, which covalently bind and inhibit the enzyme.

scholarly journals Multiplexed In Situ Protein Profiling with High-Performance Cleavable Fluorescent Tyramide

Molecules ◽  
2021 ◽  
Vol 26 (8) ◽  
pp. 2206
Author(s):  
Thai Pham ◽  
Renjie Liao ◽  
Joshua Labaer ◽  
Jia Guo
Keyword(s):  
High Performance ◽  
Single Cells ◽  
Protein Quantification ◽  
Cellular Systems ◽  
Protein Profiling ◽  
High Signal ◽  
Protein Targets ◽  
Chemical Reagents ◽  
Ffpe Tissues

Understanding the composition, function and regulation of complex cellular systems requires tools that quantify the expression of multiple proteins at their native cellular context. Here, we report a highly sensitive and accurate protein in situ profiling approach using off-the-shelf antibodies and cleavable fluorescent tyramide (CFT). In each cycle of this method, protein targets are stained with horseradish peroxidase (HRP) conjugated antibodies and CFT. Subsequently, the fluorophores are efficiently cleaved by mild chemical reagents, which simultaneously deactivate HRP. Through reiterative cycles of protein staining, fluorescence imaging, fluorophore cleavage, and HRP deactivation, multiplexed protein quantification in single cells in situ can be achieved. We designed and synthesized the high-performance CFT, and demonstrated that over 95% of the staining signals can be erased by mild chemical reagents while preserving the integrity of the epitopes on protein targets. Applying this method, we explored the protein expression heterogeneity and correlation in a group of genetically identical cells. With the high signal removal efficiency, this approach also enables us to accurately profile proteins in formalin-fixed paraffin-embedded (FFPE) tissues in the order of low to high and also high to low expression levels.

scholarly journals Insight into metastatic oral cancer tissue from novel analyses using FTIR spectroscopy and aperture IR-SNOM

The Analyst ◽  
2021 ◽  
Author(s):  
Barnaby Ellis ◽  
Conor A Whitley ◽  
Safaa Al Jedani ◽  
Caroline Smith ◽  
Philip Gunning ◽  
...  
Keyword(s):  
Machine Learning ◽  
Squamous Cell Carcinoma ◽  
Oral Cancer ◽  
Oral Squamous Cell Carcinoma ◽  
Lymphoid Tissue ◽  
Learning Algorithm ◽  
Cancer Tissue ◽  
Machine Learning Algorithm ◽  
Nodal Metastases ◽  
Insight Into

A novel machine learning algorithm is shown to accurately discriminate between oral squamous cell carcinoma (OSCC) nodal metastases and surrounding lymphoid tissue on the basis of a single metric, the...

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