scholarly journals DNA-Methylation-Mediated lncRNA HOXB-AS4 Promotes Gastric Cancer Progression Through Regulating miR-130a-5p/PKP4

2022 ◽  
Author(s):  
Xiaoyan Wang ◽  
Rong He ◽  
Yan Wang ◽  
Yunyun Liu ◽  
Yuxin Wang ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Dna Methylation ◽  
In Situ Hybridization ◽  
Western Blot ◽  
Fluorescence In Situ Hybridization ◽  
Cancer Cells ◽  
Chromatin Immunoprecipitation ◽  
Qrt Pcr ◽  
Specific Pcr

Abstract Background：Gastric cancer (GC) is one of the most common cancer in the world, possessing the second leading cause of cancer-related mortality. Long noncoding RNAs (lncRNAs) have been shown to play important roles in tumorigenesis. However, the effect of lncRNA HOXB-AS4 in GC progression and the underlying mechanisms remain unknown. Methods：Firstly, the expression of lncRNA HOXB-AS4 in gastric cancer tissues and cancer cells was investigated according to GEPIA database and Real time fluorescence quantitative PCR(qRT-PCR). Then, MTT, clone formation, Transwell and Western blot were used to study the effects of overexpression or down-regulation of HOXB-AS4 on the proliferation, invasion and epithelial mesenchymal transformation of cancer cells. We further studied the molecular mechanism of HOXB-AS4 by fluorescence in situ hybridization, bioinformatics analysis, luciferase reporting, methylation specific PCR (MSP) and chromatin immunoprecipitation (chip).Results:In the study, the GEPIA database and quantitative Real-Time PCR (qRT-PCR) assay showed that HOXB-AS4 was upregulated in GC tissues and cells. Then, MTT, clone formation, transwell, and western blot assays suggested that overexpression of HOXB-AS4 increased cell proliferation, migration, and invasion, and regulated epithelial-mesenchymal transition (EMT) markers expression, while knockdown of HOXB-AS4 showed the opposite effect. Fluorescence in situ hybridization (FISH) assay found that HOXB-AS4 localized in the cytoplasm of the GSE-1 and AGS cells. Further mechanism experiments, including bioinformatics, luciferase reporter, qRT-PCR, and western blot assays showed that HOXB-AS4 sponged to miR-130a-5p to regulate the PKP4 expression. Knockdown of miR-130a-5p obliterated the effect of HOXB-AS4, which was further abolished by knockdown of PKP4 in vitro and in vivo. Methylation-specific PCR (MSP) and chromatin immunoprecipitation (CHIP) assay showed that overexpression of HOXB-AS4 in GC was mediated by SP1-dependent DNA methylation. Abnormal upregulation of lncRNA HOXB-AS4 contributed to GC progression, which was mediated by DNA methylation. The study clarified that DNA-methylation-mediated HOXB-AS4 played its role through miR-130a-5p/PKP4 axis.Conclusions: Our study provides new insights for the understanding of epigenetic regulation on lncRNA expression in GC, and indicates that HOXB-AS4 could be a biomarker of GC prognosis. Moreover, targeting HOXB-AS4 /miR-130a-5p/PKP4 axis might be a promising strategy to treat GC.

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.

Quantitative Detection of Lung Cancer Cells by Fluorescence In Situ Hybridization

CHEST Journal ◽  
2005 ◽  
Vol 128 (2) ◽  
pp. 906-911 ◽  
Author(s):  
Haruhiko Nakamura ◽  
Idiris Aute ◽  
Norihito Kawasaki ◽  
Masahiko Taguchi ◽  
Tatsuo Ohira ◽  
...  
Keyword(s):  
Lung Cancer ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Cancer Cells ◽  
Lung Cancer Cells ◽  
Quantitative Detection

scholarly journals Combining TrkA Immunohistochemical-Score with Clinicopathologic Parameters in Neuroblastoma: An Influential Prognostic Nomogram

2021 ◽  
Author(s):  
Juanqing Yue ◽  
Lei Cai ◽  
Ruifen Wang ◽  
Meng Qiao ◽  
Kezhou Wang ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Prognostic Factor ◽  
Fluorescence In Situ Hybridization ◽  
Gene Fusion ◽  
Independent Prognostic Factor ◽  
Clinical Samples ◽  
Qrt Pcr ◽  
Clinicopathologic Parameters ◽  
Low Expression

Abstract BackgroundNeuroblastoma (NB) is one of the most common solid tumors in children with varied clinical outcomes. Although there are some several risk stratification systems currently, their clinical applications are limited due to the testing conditions of different laboratory and the heavy financial burden on patients. TrkA is coded by NTRK1 , belonging to tropomyosin receptor kinase family. We have observed that TrkA was differentially expressed in paraffin tissue sections of NB. The aim of this study was to determine the immunohistochemical-score of TrkA as an independent prognostic factor for NB and establish a useful prognostic model for postoperative patients.MethodsWe systematically summarized the relationship between immunochemistry (IHC) score of TrkA and clinicopathological parameters in 86 NB cases. Fluorescence in situ hybridization (FISH) and qRT-PCR were used to detect NTRK1 gene fusion. Furthermore, GSE96631, GSE16476, GSE49710 and GSE73537 datasets, originated from Gene Expression Omnibus (GEO), were analyzed to figure out the NTRK1 related molecular characteristics by bioinformatics methods. And combined TrkA immunohistochemical-score with clinicopathologic parameters to construct a prognostic nomogram of overall survival (OS) for NB.ResultIn clinical samples and GEO database analyses, patients in the NTRK1 / TrkA low expression group showed significantly poorer outcome than patients in high group. Multivariate cox regression analysis demonstrated NTRK1 / TrkA as an independent prognostic factor for NB survival. Neither Fluorescence in situ hybridization nor qRT-PCR detected evidence for NTRK1 gene fusion in clinical samples, indicating that differential expression in NTRK1 / TrkA are caused by epigenetic changes. Bioinformatics analyses revealed that MYC target related pathway may play a critical role in low expression of TrkA, leading to unfavorable prognoses of NB.ConclusionThe results of this study suggest that the IHC score of TrkA may be used as an independent predictor of postoperative OS of patients with NB. By combining the IHC score of TrkA and clinicopathological features, the proposed nomogram provides a feasible predictive tool for postoperative patients with NB. Simultaneously, this study also reveals that Trk inhibitors are not supposed to be taken in NB patients.

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