AKNA Is a Potential Prognostic Biomarker in Gastric Cancer and Function as a Tumor Suppressor by Modulating EMT-Related Pathways

The AT-hook transcription factor, AKNA, is a nuclear protein that affects a few physiological and pathological processes including cancer. Here, we investigated the role of AKNA in gastric cancer (GC). By using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot assays, AKNA was found deregulated in both GC cell lines and 32 paired GC tissues. Subsequently, Kaplan-Meier analysis and clinicopathological analysis were conducted using both 32 GC cases’ data above and RNA-Seq data of AKNA in 354 GC patients and the corresponding clinical-pathological data obtained from The Cancer Genome Atlas (TCGA), and AKNA expression was found closely related to location, metastasis, and TNM staging of GC. Then, the potential molecular mechanisms of AKNA in GC were explored by gene set enrichment analysis (GSEA), qRT-PCR, and Western blot assays. AKNA was found to be a hub gene related to homotypic cell to cell adhesion, regulation of cell to cell adhesion, leukocyte cell to cell adhesion, and regulation of T cell proliferation in GC. GO analysis revealed that AKNA involved in the regulation of epithelial-mesenchymal transition (EMT)-related pathways including chemokine signaling pathway, cytokine to cytokine receptor interaction, cell adhesion molecules, and jak-stat signaling pathway in GC. To explore the regulation of AKNA expression, Targetscan and TargetMiner were used to predict the possible miRNA which targeted AKNA and found the expression of AKNA was negatively correlated to miR-762 which could be sponged by circTRNC18. In conclusion, AKNA could function as a tumor suppressor by modulating EMT-related pathways in GC. The expression of AKNA might be regulated by circTRNC18/miR-762 axis. AKNA could serve as a potential biomarker and an effective target for GC diagnosis and therapy.

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Capicua homology protein inhibits the progression of gastric cancer through the PI3K/AKT signaling pathway

10.21203/rs.3.rs-29464/v1 ◽

2020 ◽

Author(s):

LU GE ◽

Chang-long Hu ◽

Zheng-hui Ge ◽

Chun-rong Wang ◽

Li Qian ◽

...

Keyword(s):

Gastric Cancer ◽

Western Blot ◽

Signaling Pathway ◽

Cell Lines ◽

Epithelial Mesenchymal Transition ◽

Akt Signaling ◽

Akt Signaling Pathway ◽

Mesenchymal Transition ◽

Matrigel Invasion ◽

Qrt Pcr

Abstract Purpose Capicua homolog protein (CIC) played a broad role in the development of cancer in humans, however, its role in the progression of gastric cancer (GC) specifically has been unclear. This study aimed to explore the expression of CIC and its potential clinical value in patients with GC. Methods The CIC levels in GC tissues and cell lines were examined by quantitative real-time polymerase chain reaction (qRT-PCR). And the in-vitro effects of CIC expression in MGC-803 cells on their proliferation, invasion, and the progression of epithelial-mesenchymal transition were assessed by CCK-8 assays, Matrigel-invasion analysis, qRT-PCR and Western blot assays, separately. In addition, the effects of downregulation of CIC on the activation of PI3K/AKT signaling pathway were measured using Western-blot analysis. Results The results showed CIC levels were lower in GC tissues and GC cell lines, and these lower CIC levels were correlated with tumor differentiation, Helicobacter pylori infection, TNM stage, and patient survival. In addition, CIC overexpression could promote cell proliferation, invasion, and progression of epithelial-mesenchymal transition in MGC-803 cells. Notably, exotic expression of CIC inactivated the phosphoinositide 3-kinase/protein kinase B signaling pathway. Conclusions In conclusion, our finding suggested CIC could serve as a potential diagnostic and prognostic biomarker and a probable therapy target for GC.

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Global transcriptomic analysis identifiesSERPINE1as a prognostic biomarker associated with epithelial-to-mesenchymal transition in gastric cancer

PeerJ ◽

10.7717/peerj.7091 ◽

2019 ◽

Vol 7 ◽

pp. e7091 ◽

Cited By ~ 5

Author(s):

Bodong Xu ◽

Zhigang Bai ◽

Jie Yin ◽

Zhongtao Zhang

Keyword(s):

Gastric Cancer ◽

Poor Prognosis ◽

Epithelial To Mesenchymal Transition ◽

Enrichment Analysis ◽

Gene Set Enrichment Analysis ◽

Plasminogen Activation ◽

Mesenchymal Transition ◽

Qrt Pcr ◽

Plasminogen Activator System ◽

Km Plotter

BackgroundThe plasminogen activation system plays a pivotal role in regulating tumorigenesis. In this work, we aim to identify key regulators of plasminogen activation associated with tumorigenesis and explore potential mechanisms in gastric cancer (GC).MethodsGene profiling datasets were extracted from the Gene Expression Omnibus (GEO) database. The differentially expressed genes (DEGs) were screened for and obtained by the GEO2R tool. The Database for Annotation, Visualization and Integrated Discovery was used for GO and KEGG enrichment analysis. Gene set enrichment analysis (GSEA) was performed to verify molecular signatures and pathways among The Cancer Genome Atlas or GEO datasets. Correlations between SERPINE1 and markers of epithelial-to-mesenchymal transition (EMT) were analyzed using the GEPIA database and quantitative real-time PCR (qRT-PCR). Interactive networks of selected genes were built by STRING and Cytoscape software. Finally, selected genes were verified with the Kaplan–Meier (KM) plotter database.ResultsA total of 104 overlapped upregulated and 61 downregulated DEGs were obtained. Multiple GO and KEGG terms associated with the extracellular matrix were enriched among the DEGs. SERPINE1 was identified as the only regulator of angiogenesis and the plasminogen activator system among the DEGs. A high level of SERPINE1 was associated with a poor prognosis in GC. GSEA analysis showed a strong correlation between SERPINE1 and EMT, which was also confirmed with the GEPIA database and qRT-PCR validation. FN1, TIMP1, MMP2, and SPARC were correlated with SERPINE1.The KM plotter database showed that an overexpression of these genes correlated with a shorter survival time in GC patients.ConclusionsIn conclusion, SERPINE1 is a potent biomarker associated with EMT and a poor prognosis in GC. Furthermore, FN1, TIMP1, MMP2, and SPARC are correlated with SERPINE1 and may serve as therapeutic targets in reversing EMT in GC.

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MiR-92a-3p promotes the malignant progression of hepatocellular carcinoma by mediating the PI3K/AKT/mTOR signaling pathway

Current Pharmaceutical Design ◽

10.2174/1381612827666210612054156 ◽

2021 ◽

Vol 27 ◽

Author(s):

Libing Wang ◽

Mingxin Cui ◽

Fengzhi Qu ◽

Daming Cheng ◽

Jingkun Yu ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Western Blot ◽

Signaling Pathway ◽

Epithelial Mesenchymal Transition ◽

Mtor Signaling ◽

Mtor Signaling Pathway ◽

Mesenchymal Transition ◽

Diagnosis And Prognosis ◽

Potential Biomarker ◽

Hcc Cells

Background: As one of the most common cancers in the world, hepatocellular carcinoma (HCC) usually has a poor prognosis. Many HCC patients are usually diagnosed at advanced stages. Therefore, new potential biomarkers for the diagnosis and prognosis of HCC are urgently needed. More and more studies have shown that miR-92a-3p can regulate the occurrence and development of a variety of cancers, but its clinical significance and molecular mechanism in HCC are still elusive. Here, we tried to clarify the regulatory mechanism of miR-92a-3p in HCC. Methods: In this study, we conducted qRT-PCR and revealed that miR-92a-3p was notably upregulated in HCC cells. MTT, flow cytometry, wound healing, Transwell invasion assays and western blot were conducted to uncover that overexpressed miR-92a-3p could boost the proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) of HCC cells while inhibiting cell apoptosis. In addition, the proteins associated with PI3K/AKT/mTOR pathway were also detected by western blot. Results: It was suggested that miR-92a-3p could activate the PI3K/AKT/mTOR signaling pathway. Conclusion: These results suggest that miR-92a-3p plays a tumor-promoting role in HCC and may be a potential biomarker for the diagnosis and prognosis of HCC.

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MicroRNA-491-5p inhibit gastric cancer development by targeting EMT, cell adhesion genes and IFITM2

10.21203/rs.2.20605/v2 ◽

2020 ◽

Author(s):

zhijian Wei ◽

lixiang Zhang ◽

angqing Li ◽

chuanhong li ◽

wenxiu han ◽

...

Keyword(s):

Gastric Cancer ◽

Cell Adhesion ◽

Tumor Suppressor ◽

Epithelial Mesenchymal Transition ◽

Migration And Invasion ◽

Western Blot Assay ◽

Mesenchymal Transition ◽

High Level

Abstract Background: Gastric cancer (GC) is one of the deadliest cancers in China. And, it can be regulated by MicroRNAs (miRNAs) generally. miR-491-5p function as a tumor suppressor in different types of cancer, but we still don’t know the role of miR-491-5p in gastric cancer. Methods: Functional experiments including CCK-8 assay and transwell assay were performed. Furthermore, the underlying mechanism was explored through qRT-PCR and western blot assay. In addition, the function of miR-491-5p was also identified in vivo.Results: In this study, we found that high level of miR-491-5p caused a weak cell proliferation, migration and invasion abilities. In order to explore the role of miR-491-5p in vivo, we set a xenograft mouse model, and also found that high level of miR-491-5p suppressed tumor growth. Moreover, we found that miR-491-5p regulate the tumor development thought regulate the expression of EMT(Epithelial-mesenchymal transition), cell adhesion genes and IFITM2. Conclusions: These data show that miR-491-5p function as a tumor suppressor in GC both in vitro and in vivo.

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ANXA1 Induces Oxaliplatin Resistance of Gastric Cancer through Autophagy by Targeting PI3K/AKT/mTOR

10.21203/rs.3.rs-895675/v1 ◽

2021 ◽

Author(s):

Jun Ren ◽

Qing Zhi Hu ◽

Ming Geng Niu ◽

Jie Xia ◽

Xing Wang ◽

...

Keyword(s):

Gastric Cancer ◽

Western Blot ◽

Signaling Pathway ◽

Enrichment Analysis ◽

Mtor Signaling ◽

Gene Set Enrichment Analysis ◽

Rna Seq ◽

Mtor Signaling Pathway

Abstract Background: Resistance to oxaliplatin (OXA) is a major cause of recurrence in gastric cancer (GC) patients. ANXA1 has been found to participate in the regulation of diverse cellular functions in a variety of cell types including anti-inflammatory processes. We aimed to investigate the role of ANXA1 in autophagy and chemoresistance of GC cells. Methods: To identify the genes that regulate oxaliplatin resistance, we used RNA-seq to profile gene expression within oxaliplatin resistant GC and parental cells. Immunohistochemical and RT-qPCR was performed to detect ANXA1 expression in tissues of 2 cohorts of GC patients who received OXA-based chemotherapy. The chemoresistant effects of ANXA1 were assessed by cell viability, apoptosis, and autophagy assays. The effects of ANXA1 on autophagy were assessed by mRFP-GFP-LC3 and western blot. Gene set enrichment analysis (GSEA) and western blot was performed to detect the activity of PI3K/AKT/mTOR signaling under the regulation of ANXA1.Results: Based on RNA-seq profiling, ANXA1 was selected as a candidate that was upregulated in oxaliplatin resistant GC cells. Furthermore, we discovered that ANXA1 is upregulated in chemo-resistant GC tissues. Knockdown of ANXA1, via inhibiting autophagy, enhanced the sensitivity of OXA-resistant GC cells to OXA in vitro and in vivo. Mechanically, we identified that PI3K/AKT/mTOR signaling pathway was activated in the ANXA1 stably knockdown AGS/OXA cells, which leads to the down-regulation of autophagy.Conclusions: ANXA1 functions as a chemoresistant gene in GC cells by targeting the PI3K/AKT/mTOR signaling pathway and might be a prognostic predictor for GC patients who receive OXA-based chemotherapy.

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Genome-Wide Lymphocytic mRNA Sequencing to Identify Epithelial-Mesenchymal Transition Mechanism in Silicosis

10.21203/rs.3.rs-1046313/v1 ◽

2021 ◽

Author(s):

Hai bin Li ◽

Lin Zhang ◽

Yi Guan ◽

Yingzheng Zhao ◽

Ning Li ◽

...

Keyword(s):

Cell Adhesion ◽

Signaling Pathway ◽

Peripheral Blood ◽

Epithelial Mesenchymal Transition ◽

Janus Kinase ◽

Gene Set Enrichment Analysis ◽

Healthy Controls ◽

Blood Samples ◽

Mesenchymal Transition ◽

Transition Mechanism

Abstract Background Lymphocytes are immune cells that play dual roles in the pathogenesis of silicosis. Epithelial-mesenchymal transition (EMT), a vital phenomenon in the pathogenesis of silicosis, is regulated by cytokines, chemokines, and other molecules secreted by lymphocytes; however, the underlying regulatory mechanism is unclear. Here, we investigated the role of lymphocytes in EMT in silicosis. Methods Three patients with silicosis and three healthy controls that underwent pre-job physical examination were recruited; fasting venous blood samples were collected and lymphocytes were separated by Ficoll. High-throughput sequencing technology and bioinformatic analysis were used to identify specific genes and signaling pathways. The results were verified through the detection of related indices of peripheral blood samples. Results The baseline characteristics of subjects from silicosis group were matched with those of healthy controls. In comparison with healthy controls, patients with silicosis showed 1915 dysregulated genes that were thought to participate in various biological processes, including angiogenesis, tissue repair, cell proliferation, invasion, migration, and EMT. Protein-protein interaction analysis grouped these genes into three hub targets, including phosphoinositide 3-kinase (PI3K), integrin beta 1 (ITGB1), and integrin-linked protein kinase (ILK). Gene set enrichment analysis (GSEA) confirmed that PI3K, ITGB1, and ILK were tightly associated with EMT through the Wnt signaling pathway, Janus kinase/signal transducer and activator of transcription (JAK-STAT) signaling pathway, and cell adhesion molecular pathway. ITGB1 is a member of the adhesion molecule family. The identified genes were verified through the detection of soluble adhesion molecules in peripheral blood samples of patients with silicosis and healthy subjects. Conclusion Dysregulation of PI3K, ITGB1, and ILK in lymphocytes may contribute to EMT via JAK-STAT, Wnt, and cell adhesion molecular pathways in patients with silicosis.

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Ginsenoside Rg3 Inhibits the Growth of Osteosarcoma and Attenuates Metastasis through the Wnt/β-Catenin and EMT Signaling Pathway

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2020/6065124 ◽

2020 ◽

Vol 2020 ◽

pp. 1-12

Author(s):

Xiaohan Mao ◽

Yaqian Jin ◽

Tianyu Feng ◽

Hao Wang ◽

Dan Liu ◽

...

Keyword(s):

Western Blot ◽

Signaling Pathway ◽

Epithelial Mesenchymal Transition ◽

Chinese Herb ◽

Luciferase Reporter ◽

Migration And Invasion ◽

Ginsenoside Rg3 ◽

Mesenchymal Transition ◽

Qrt Pcr ◽

Gene Assay

Osteosarcoma (OS) is the most common primary malignant bone cancer. An increasing number of studies have demonstrated that ginsenoside Rg3 (Rg3), which is extracted from the roots of the traditional Chinese herb Panax ginseng, plays a tumor suppression role in various malignant tumors. In the present study, we aimed at investigating the role of Rg3 in the proliferation, migration, and invasion of OS and at exploring the underlying mechanisms. Cell viability and proliferation were observed by MTT assay and crystal violet staining. The migration and invasion of cells were measured by wound-healing assay and Transwell method. Signaling pathway screening was investigated using luciferase reporter gene assay. qRT-PCR and western blot were performed to measure the expression of molecules involved in cell epithelial-mesenchymal transition (EMT), and Wnt/β-catenin pathway. Results suggested that Rg3 could not only inhibit proliferation but also hamper the migration and invasion of OS. qRT-PCR and western blot demonstrated that a reduced level of MMP2/MMP7/MMP9 was induced after Rg3 treatment. In addition, the expression levels of proteins related to EMT and the Wnt/β-catenin pathway were downregulated. In summary, our data revealed that Rg3 could inhibit the proliferation, migration, and invasion of OS cells. This effect of Rg3 might be mediated by downregulating MMP2, MMP7, and MMP9 expression and suppressing EMT as well as the Wnt/β-catenin pathway. Thus, Rg3 might be a potential agent for the treatment of OS.

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Integrated Multiomics Analysis Identifies a Novel Biomarker Associated with Prognosis in Intracerebral Hemorrhage

Oxidative Medicine and Cellular Longevity ◽

10.1155/2021/2510847 ◽

2021 ◽

Vol 2021 ◽

pp. 1-20

Author(s):

Yiqing Shen ◽

Wensong Yang ◽

Xin Xiong ◽

Xinhui Li ◽

Zhongsong Xiao ◽

...

Keyword(s):

Brain Injury ◽

Intracerebral Hemorrhage ◽

Western Blot ◽

Gene Set Enrichment Analysis ◽

Multidimensional Analysis ◽

Differentially Expressed ◽

Secondary Brain Injury ◽

Hub Genes ◽

Qrt Pcr ◽

Potential Biomarker

Existing treatments for intracerebral hemorrhage (ICH) are unable to satisfactorily prevent development of secondary brain injury after ICH and multiple pathological mechanisms are involved in the development of the injury. In this study, we aimed to identify novel genes and proteins and integrated their molecular alternations to reveal key network modules involved in ICH pathology. A total of 30 C57BL/6 male mice were used for this study. The collagenase model of ICH was employed, 3 days after ICH animals were tested neurological. After it, animals were euthanized and perihematomal brain tissues were collected for transcriptome and TMT labeling-based quantitative proteome analyses. Protein-protein interaction (PPI) network, Gene Set Enrichment Analysis (GSEA), and regularized Canonical Correlation Analysis (rCCA) were performed to integrated multiomics data. For validation of hub genes and proteins, qRT-PCR and Western blot were carried out. The candidate biomarkers were further measured by ELISA in the plasma of ICH patients and the controls. A total of 2218 differentially expressed genes (DEGs) and 353 differentially expressed proteins (DEPs) between the ICH model group and control group were identified. GSEA revealed that immune-related gene sets were prominently upregulated and significantly enriched in pathways of inflammasome complex, negative regulation of interleukin-12 production, and pyroptosis during the ICH process. The rCCA network presented two highly connective clusters which were involved in the sphingolipid catabolic process and inflammatory response. Among ten hub genes screened out by integrative analysis, significantly upregulated Itgb2, Serpina3n, and Ctss were validated in the ICH group by qRT-PCR and Western blot. Plasma levels of human SERPINA3 (homologue of murine Serpina3n) were elevated in ICH patients compared with the healthy controls (SERPINA3: 13.3 ng/mL vs. 11.2 ng/mL, p = 0.015 ). Within the ICH group, higher plasma SERPINA3 levels with a predictive threshold of 14.31 ng/mL ( sensitivity = 64.3 % ; specificity = 80.8 % ; AUC = 0.742 , 95% CI: 0.567-0.916) were highly associated with poor outcome (mRS scores 4-6). Taken together, the results of our study exhibited molecular changes related to ICH-induced brain injury by multidimensional analysis and effectively identified three biomarker candidates in a mouse ICH model, as well as pointed out that Serpina3n/SERPINA3 was a potential biomarker associated with poor functional outcome in ICH patients.

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Bioinformatics Analysis Identifies CPZ as a Tumor Immunology Biomarker For Gastric Cancer

Current Bioinformatics ◽

10.2174/1574893615999200707145643 ◽

2020 ◽

Vol 15 ◽

Author(s):

Yuan Gu ◽

Ying Gao ◽

Xiaodan Tang ◽

Huizhong Xia ◽

Kunhe Shi

Keyword(s):

Gastric Cancer ◽

T Cells ◽

Signaling Pathway ◽

Tumor Immunology ◽

Bioinformatics Analysis ◽

Human Cancer ◽

Disease Free Survival ◽

Kaplan Meier ◽

Potential Biomarker ◽

Pathway Regulation

Background: Gastric cancer (GC) is one of the most common malignancies worldwide. However, the biomarkers for the prognosis and diagnosis of Gastric cancer were still need. Objective: The present study aimed to evaluate whether CPZ could be a potential biomarker for GC. Method: Kaplan-Meier plotter (http://kmplot.com/analysis/) was used to determine the correlation between CPZ expression and overall survival (OS) and disease-free survival (DFS) time in GC [9]. We analyzed CPZ expression in different types of cancer and the correlation of CPZ expression with the abundance of immune infiltrates, including B cells, CD4+ T cells, CD8+ T cells, neutrophils, macrophages, and dendritic cells, via gene modules using TIMER Database. Results: The present study identified that CPZ was overexpressed in multiple types of human cancer, including Gastric cancer. We found that overexpression of CPZ correlates to the poor prognosis of patients with STAD. Furthermore, our analyses show that immune infiltration levels and diverse immune marker sets are correlated with levels of CPZ expression in STAD. Bioinformatics analysis revealed that CPZ was involved in regulating multiple pathways, including PI3K-Akt signaling pathway, cGMP-PKG signaling pathway, Rap1 signaling pathway, TGF-beta signaling pathway, regulation of cell adhesion, extracellular matrix organization, collagen fibril organization, collagen catabolic process. Conclusion: This study for the first time provides useful information to understand the potential roles of CPZ in tumor immunology and validate it to be a potential biomarker for GC.

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Interrogation of gender disparity uncovers androgen receptor as the transcriptional activator for oncogenic miR-125b in gastric cancer

Cell Death and Disease ◽

10.1038/s41419-021-03727-3 ◽

2021 ◽

Vol 12 (5) ◽

Author(s):

Ben Liu ◽

Meng Zhou ◽

Xiangchun Li ◽

Xining Zhang ◽

Qinghua Wang ◽

...

Keyword(s):

Gastric Cancer ◽

Androgen Receptor ◽

Signaling Pathway ◽

Gender Bias ◽

Gender Disparity ◽

Gene Set Enrichment Analysis ◽

Cancer Type ◽

Female Patients

AbstractThere is a male preponderance in gastric cancer (GC), which suggests a role of androgen and androgen receptor (AR). However, the mechanism of AR signaling in GC especially in female patients remains obscure. We sought to identify the AR signaling pathway that might be related to prognosis and examine the potential clinical utility of the AR antagonist for treatment. Deep learning and gene set enrichment analysis was used to identify potential critical factors associated with gender bias in GC (n = 1390). Gene expression profile analysis was performed to screen differentially expressed genes associated with AR expression in the Tianjin discovery set (n = 90) and TCGA validation set (n = 341). Predictors of survival were identified via lasso regression analyses and validated in the expanded Tianjin cohort (n = 373). In vitro and in vivo experiments were established to determine the drug effect. The GC gender bias was attributable to sex chromosome abnormalities and AR signaling dysregulation. The candidates for AR-related gene sets were screened, and AR combined with miR-125b was associated with poor prognosis, particularly among female patients. AR was confirmed to directly regulate miR-125b expression. AR-miR-125b signaling pathway inhibited apoptosis and promoted proliferation. AR antagonist, bicalutamide, exerted anti-tumor activities and induced apoptosis both in vitro and in vivo, using GC cell lines and female patient-derived xenograft (PDX) model. We have shed light on gender differences by revealing a hormone-regulated oncogenic signaling pathway in GC. Our preclinical studies suggest that AR is a potential therapeutic target for this deadly cancer type, especially in female patients.

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