scholarly journals Identification of miR-589-5p as a Potential Prognostic Biomarker of Hepatocellular Carcinoma

2021 ◽  
Author(s):  
Zhijia Yao ◽  
Chunlin Jiang ◽  
Huiwen Weng ◽  
Shi Fang ◽  
Heping Li ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
In Situ Hybridization ◽  
Early Diagnosis ◽  
Real Time ◽  
Real Time Pcr ◽  
Human Cancer ◽  
Clinical Stage ◽  
Survival Prognosis ◽  
Expression Levels

Abstract Background: MicroRNAs (miRNAs) are small endogenous non-coding RNAs with 22 nucleotides approximately. Numerous studies have reported that microRNAs may hold the potential to serve as biomarkers in diagnosing cancer and predicting prognosis. miR-589-5p is a well-documented oncogenic miRNA implicated in several human cancer types. However, the potential value of miR-589-5p as a biomarker to facilitate early diagnosis and predict prognosis in hepatocellular carcinoma (HCC) patients remains obscure.Methods: miR-589-5p expression was examined by in situ hybridization (ISH) in 10 adjacent normal tissues (ANT), 21 liver fibrosis tissues, 113 HCC tissues, including 22 HCC with grade I, 37 HCC with grade II, 49 HCC with grade III and 5 hepatocholangiocellular carcinoma tissues. Statistical analysis and Kaplan–Meier survival analysis was performed to evaluate the clinical correlation between miR-589-5p expression and clinicopathological features and survival prognosis in HCC patients.Results: miR-589-5p expression was dramatically upregulated in HCC tissues by in situ hybridization (ISH). Overexpression of miR-589-5p was not only positively correlated with poor tumor differentiation degree, increased AFP levels, advanced clinical stage, distant metastatic status and venous invasion, but also predicted poor overall and relapse-free survival in HCC patients. Importantly, the expression levels of miR-589-5p measured by real-time PCR in high-ISH scores tissues were dramatically upregulated compared with those in low-ISH scores tissues, and a strong and positive correlation of miR-589-5p expression levels in real-time PCR with ISH staining index was demonstrated in HCC tissues.Conclusion: our results support the notion that miR-589-5p might hold a promising value as a novel biomarker to facilitate early diagnosis and predict prognosis in HCC patients.

Quantification of an Eikelboom type 021N bulking event with fluorescence in situ hybridization and real-time PCR

2005 ◽  
Vol 68 (5) ◽  
pp. 695-704 ◽  
Author(s):  
Han Vervaeren ◽  
Kurt De Wilde ◽  
Jorg Matthys ◽  
Nico Boon ◽  
Lutgarde Raskin ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Real Time ◽  
Real Time Pcr

scholarly journals OCT4B1 Regulates the Cellular Stress Response of Human Dental Pulp Cells with Inflammation

2017 ◽  
Vol 2017 ◽  
pp. 1-10 ◽  
Author(s):  
Lu Liu ◽  
Rong Huang ◽  
Ruiqi Yang ◽  
Xi Wei
Keyword(s):  
In Situ Hybridization ◽  
Survival Rate ◽  
Western Blot ◽  
Real Time ◽  
Real Time Pcr ◽  
Dental Pulp ◽  
Immunofluorescence Staining ◽  
Dental Pulp Cells ◽  
Pulp Cells

Introduction. Infection and apoptosis are combined triggers for inflammation in dental tissues. Octamer-binding transcription factor 4-B1 (OCT4B1), a novel spliced variant of OCT4 family, could respond to the cellular stress and possess antiapoptotic property. However, its specific role in dental pulpitis remains unknown. Methods. To investigate the effect of OCT4B1 on inflammation of dental pulp cells (DPCs), its expression in inflamed dental pulp tissues and DPCs was examined by in situ hybridization, real-time PCR, and FISH assay. OCT4B1 overexpressed DPCs model was established, confirmed by western blot and immunofluorescence staining, and then stimulated with Lipopolysaccharide (LPS). Apoptotic rate was determined by Hoechst/PI staining and FACS. Cell survival rate was calculated by CCK8 assay. Results. In situ hybridization, real-time PCR, and FISH assay revealed that OCT4B1 was extensively expressed in inflamed dental pulp tissues and DPCs with LPS stimulation. Western blot and immunofluorescence staining showed the expression of OCT4B1 and OCT4B increased after OCT4B1 transfection. Hoechst/PI staining and FACS demonstrated that less red/blue fluorescence was detected and apoptotic percentage decreased (3.45%) after transfection. CCK8 demonstrated that the survival rate of pCDH-OCT4B1-flag cells increased. Conclusions. OCT4B1 plays an essential role in inflammation and apoptosis of DPCs. OCT4B might operate synergistically with OCT4B1 to reduce apoptosis.

scholarly journals Identification of immune-infiltrating cell-related biomarkers in hepatocellular carcinoma based on gene co-expression network analysis

2021 ◽  
Vol 16 (1) ◽  
Author(s):  
Yinghui Hou ◽  
Guizhi Zhang
Keyword(s):  
Hepatocellular Carcinoma ◽  
Network Analysis ◽  
Early Diagnosis ◽  
The Cancer Genome Atlas ◽  
Ppi Network ◽  
Sequencing Data ◽  
Hub Genes ◽  
Survival Prognosis ◽  
Expression Levels ◽  
Cancer Tissues

Abstract Background Hepatocellular carcinoma (HCC) is often caused by chronic liver infection or inflammation. Searching for potential immunotherapy targets will aid the early diagnosis and treatment of HCC. Methods Firstly, detailed HCC data were downloaded from The Cancer Genome Atlas database. GDCRNATools was used for the comprehensive analysis of RNA sequencing data. Subsequently, the CIBERSORT package was used to estimate infiltration scores of 22 types of immune cells in complex samples. Furthermore, hub genes were identified via weighted gene co-expression network analysis (WGCNA) and protein-protein interaction (PPI) network analysis. In addition, multiple databases were used to validate the expression of hub gene in the tumor tissue. Finally, prognostic, diagnostic and immunohistochemical analysis of key hub genes was performed. Results In the present study, 9 hub genes were identified using WGCNA and PPI network analysis. Furthermore, the expression levels of 9 genes were positively correlated with the infiltration levels of CD8-positive T (CD8+ T) cells. In multiple dataset validations, the expression levels of CCL5, CXCR6, CD3E, and LCK were decreased in cancer tissues. In addition, survival analysis revealed that patients with LCK low expression had a poor survival prognosis (P < 0.05). Immunohistochemistry results demonstrated that CCL5, CD3E and LCK were expressed at low levels in HCC cancer tissues. Conclusion The identification of CCL5, CXCR6, CD3E and LCK may be helpful in the development of early diagnosis and therapy of HCC. LCK may be a potential prognostic biomarker for immunotherapy for HCC.

scholarly journals Histological Lesions and Replication Sites of PCV3 in Naturally Infected Pigs

Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1520
Author(s):  
Elisa Rigo De Conti ◽  
Talita Pilar Resende ◽  
Lacey Marshall-Lund ◽  
Albert Rovira ◽  
Fabio Augusto Vannucci
Keyword(s):  
In Situ Hybridization ◽  
Lymph Nodes ◽  
Real Time ◽  
Real Time Pcr ◽  
Systemic Vasculitis ◽  
Tissue Type ◽  
Lymphoid Tissues ◽  
Associated Diseases ◽  
Spleen And Lymph Nodes

Porcine circovirus type 3 (PCV3) has been recently described as a potential cause of abortions and systemic vasculitis in pigs. Although the virus has been detected by real-time PCR in several porcine tissues from countries worldwide, PCV3-associated diseases have not been satisfactorily clarified. The objective of this study was to investigate the association between the presence of PCV3 mRNA detected by in situ hybridization (ISH) within histological lesions and PCV3 DNA detected by real-time PCR in naturally infected pigs. A total of 25 PCV3 PCR-positive cases were analyzed. Formalin-fixed tissues from these cases were evaluated for histologic lesions and for ISH-RNA positive signals for PCV3. The most frequent tissue type with histopathologic lesions was heart, 76.2%, with lymphoplasmacytic myocarditis and epicarditis as the most frequent lesions observed. Lymphoplasmacytic interstitial pneumonia was also a frequent finding, 47.6%. There were also lesions in kidney, liver, spleen and lymph nodes. PCV3-ISH-RNA positive signals were mostly observed in association with lymphoplasmacytic inflammatory infiltrate in various tissues, including arteries. Based on our results, the minimum set of specimens to be submitted for histopathology and mRNA in situ hybridization to confirm or exclude a diagnosis of PCV3 are heart, lung and lymphoid tissues (i.e., spleen and lymph nodes), especially for differential diagnosis related with PCV2-associated diseases.

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