Rs56288038 (C/G) in 3'UTR of IRF-1 Regulated by MiR-502-5p Promotes Gastric Cancer Development

Background/Aims: Interferon regulatory factor 1 (IRF-1) has been shown to function as a transcriptional activator or repressor of a variety of target genes. However, its upstream, non-coding RNA-related regulatory capacity remains unknown. In this study, we focus on the miRNA-associated single nucleotide polymorphisms (SNPs) in the 3′untranslated region (UTR) of IRF-1 to further investigate the functional relationship and potential diagnostic value of the SNPs and miRNAs among Chinese gastric cancer (GC) patients. Methods: We performed a case-control study with 819 GC patients and 756 cancer-free controls. Genotyping by realtime PCR assay, cell transfection, and the dual luciferase reporter assay were used in our study, and the 5-year overall survival rate and relapse-free survival rate in different groups were investigated. Results: We found that patients suffering from Helicobacter pylori (Hp) infection were the susceptible population compared to controls. SNP rs56288038 (C/G) in IRF-1 3′UTR was involved in the occurrence of GC by acting as a tumor promoter factor. SNP rs56288038 (C/G) could be up-regulated by miR-502-5p, which caused a down-regulation of IRF-1 in cell lines and decreased apoptosis induced by IFN-γ. Carrying the G genotype was related to significantly low expression of IRF-1 and Hp infection, poor differentiation, big tumor size, invasion depth, as well as the high probability of metastasis, and moreover, the C/G SNP was associated with shorter survival of GC patients with five years of follow-up study. Conclusions: our findings have shown that the SNP rs56288038 (C/G) in IRF-1 3′UTR acted as a promotion factor in GC development through enhancing the regulatory role of miR-502-5p in IRF-1 expression.

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Relationship between miR-378c and YY1 expression in patients with gastric cancer and the clinicopathological features

Cellular & Molecular Biology Letters ◽

10.1186/s11658-021-00256-x ◽

2021 ◽

Vol 26 (1) ◽

Author(s):

Lei Zhang ◽

Lei Zou ◽

Peng Sun

Keyword(s):

Gastric Cancer ◽

Prognostic Factors ◽

Survival Rate ◽

Diagnostic Value ◽

Tnm Staging ◽

Luciferase Reporter Assay ◽

Luciferase Reporter ◽

Reporter Assay ◽

Low Expression ◽

Dual Luciferase Reporter Assay

Abstract Background The purpose of this study was to explore the clinical value of miR-378c and its target gene YY1 in gastric cancer. Methods The TCGA database was employed to analyse miR-378c expression in gastric cancer. qRT-PCR was applied to identify miR-378c and YY1 in tissues and serum of patients suffering from gastric cancer. The association of miR-378c with the clinical data of patients with gastric cancer was analysed. Receiver operating characteristics (ROC) curve analysis was used to determine the diagnostic value of miR-378c and YY1 in gastric cancer, and analyse the relationship between miR-378c and YY1 and patients’ survival. Pearson’s test was applied to determine the association between miR-378c and YY1 in tissue and serum of patients. Dual-Luciferase Reporter assay was employed to examine the targeting association between miR-378c and YY1. Finally, independent prognostic factors was determined in patients with gastric cancer using Cox regression analysis. Results In the TCGA database, miR-378c was weakly expressed in gastric cancer. Overall, patients with low expression had a lower survival rate. The expression of miR-378c decreased and the expression of YY1 increased in cancer tissues and serum of tumour patients. In patients with low expression of miR-378c the tumour size was ≥ 5 cm. Low differentiation, high TNM staging and lymph node invasion rate increased significantly, but the 5-year survival rate decreased in the patients. miR-378c and YY1 had better diagnostic value in gastric cancer. TargetScan, miRDB, starBase and miRTarBase predicted that YY1 was a potential gene of miR-378c, and the Dual-Luciferase Reporter assay revealed that there was a targeting relationship between the two, which was proved by correlation analysis. Multivariate Cox analysis revealed that differentiation, TNM staging and miR-378c were independent prognostic factors for patients. Conclusions MiR-378c is weakly expressed in gastric cancer patients and may be considered as a promising diagnostic and prognostic indicator for gastric cancer.

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The Functional SOCS3 RS115785973 Variant Regulated by MiR-4308 Promotes Gastric Cancer Development in Chinese Population

Cellular Physiology and Biochemistry ◽

10.1159/000443118 ◽

2016 ◽

Vol 38 (5) ◽

pp. 1796-1802 ◽

Cited By ~ 10

Author(s):

Xiaowei Wang ◽

Ting Li ◽

Meng Li ◽

Na Cao ◽

Jun Han

Keyword(s):

Gastric Cancer ◽

Risk Factor ◽

Luciferase Reporter ◽

Vitro Assay ◽

Susceptible Population ◽

Study Results ◽

Socs3 Expression ◽

Poor Differentiation ◽

H Pylori

Background/Aims: SOCS3 is tumor suppressor which has been identified as upstream of JAK/STAT3 signaling by specific kinase inhibition. However, additional regulations especially through a non-coding RNA approach were remained unknown. Methods: We performed case-control study focusing on the miRNAs associated SNPs in SOCS3 to investigate the further relationship of the SNPs with miRNAs among Chinese gastric cancer (GC) patients. Genotyping, real time PCR assay, cell transfection and the dual luciferase reporter assay were used in our study. Results: We found that patients suffering from Helicobacter pylori (H. pylori) infection indicating as susceptible population by comparing with controls. Besides, SNP rs115785973 in SOCS3 was identified as a risk factor in the occurrence of GC highly associated with poor differentiation grade, larger tumor size and metastasis. In vitro assay found that rs115785973 could be regulated by miR-4308 which caused an up-regulation of SOCS3 in patients with GA and AA genotype. Conclusion: Our findings have shown that the SNP rs115785973 in SOCS3 disrupting the regulatory role of miR-4308 in SOCS3 expression, rs115785973 in SOCS3 might act as a risk factor in the pathogenesis of GC.

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LncRNA HCG11 promotes proliferation and migration in gastric cancer via targeting miR-1276/CTNNB1 and activating Wnt signaling pathway

Cancer Cell International ◽

10.1186/s12935-019-1046-0 ◽

2019 ◽

Vol 19 (1) ◽

Cited By ~ 6

Author(s):

Hua Zhang ◽

Haitao Huang ◽

Xiaomei Xu ◽

Haiying Wang ◽

Jianxiang Wang ◽

...

Keyword(s):

Gastric Cancer ◽

Wnt Signaling ◽

Signaling Pathway ◽

Function Analysis ◽

Wnt Signaling Pathway ◽

Tumor Formation ◽

Tumor Promoter ◽

Luciferase Reporter ◽

And Migration ◽

Proliferation And Migration

Abstract Background Gastric cancer (GC) is one common cancer which occurs in the stomach leading to high mortality around the world. Long non-coding RNAs (lncRNAs) were found overexpressed or silenced in the occurrence and progression of multiple cancers including GC. Method The gene expression level in GC tissues and cells were analyzed by RT-qPCR. CCK-8, colony formation, flow cytometry and transwell assays were performed for the function analysis of HLA complex group 11 (HCG11). The mechanism study for HCG11 was conducted using RIP, RNA pull down and luciferase reporter assays. Results HCG11 was discovered highly expressed in GC tissues and cells. Depletion experiments were used to evaluate HCG11 silence on cell proliferation, migration and apoptosis. Moreover, Wnt signaling pathway was found as a tumor promoter in GC. RIP assay, RNA pull down assay and luciferase reporter assay were performed to illustrate the relationship of HCG11, miR-1276 and CTNNB1. Rescue assays revealed that HCG11/miR-1276/CTNNB1 axis regulated the incidence and development of GC. Tumor formation in mice proved that HCG11 was negatively correlated with miR-1276 and had positively correlation with CTNNB1. Conclusion Overall, HCG11 accelerated proliferation and migration in GC through miR-1276/CTNNB1 and Wnt signaling pathway, revealing that HCG11 could be a brand new target for GC.

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LINC00511 accelerated the process of gastric cancer by targeting miR-625-5p/NFIX axis

Cancer Cell International ◽

10.1186/s12935-019-1070-0 ◽

2019 ◽

Vol 19 (1) ◽

Cited By ~ 8

Author(s):

Zhaosheng Chen ◽

Honglei Wu ◽

Zhen Zhang ◽

Guangchun Li ◽

Bin Liu

Keyword(s):

Gastric Cancer ◽

Binding Sites ◽

Regulatory System ◽

Tumor Promoter ◽

Luciferase Reporter ◽

Loss Of Function ◽

Ki67 Expression ◽

Downstream Target ◽

Reporter Assays ◽

Non Coding Rnas

Abstract Background Gastric cancer (GC) is a common-sighted cancer which is hard to cure over the world. Substantial researches revealed that long non-coding RNAs (lncRNAs) were fundamental regulators in the process of cancers. Nevertheless, the biological function of LINC00511 and how LINC00511 was involved in the regulatory system in GC remained unclear. Methods RIP assays and luciferase reporter assays were performed to illustrate combination between LINC00511 and miR-625-5p. Loss-of-function assays were applied for identifying LINC00511 function in GC. Results In our study, LINC00511 was discovered significantly high in expression in GC tissues and cell lines. Moreover, LINC00511 showed a strong expression in I/II and III/IV stage. Knockdown of LINC00511 could inhibit the cell proliferation while enhanced cell apoptosis rate in GC. We used nuclear–cytoplasmic fractionation to judge the subcellular localization of LINC00511. Furthermore, miR-625-5p was found to have binding sites for LINC00511 and negatively regulated by LINC00511. Overexpression of miR-625-5p repressed the course of GC. And knockdown of miR-625-5p could recover the effects of LINC00511 silence. Besides, NFIX was discovered as a downstream target of miR-625-5p and overexpression of NFIX could offset the influence of LINC00511 silence. The results of vivo studies manifested that down-regulation of LINC00511 could reduce the Ki67 expression and NFIX while lifted the expression of miR-625-5p. Conclusion Overall, the results from our study demonstrated that LINC00511 could function as a tumor promoter by targeting miR-625-5p NFIX axis, suggesting LINC00511 could be considered as a target for GC treatment.

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miR-3613 Regulates Gastric Cancer Cells Proliferation, Invasion, Migration and Apoptosis by Targeting Suppressor of Cytokine Signaling 4

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2019.22051699 ◽

2019 ◽

Vol 9 (12) ◽

pp. 1699-1705

Author(s):

Yuming Luo ◽

Wei Cao

Keyword(s):

Gastric Cancer ◽

Cell Cycle ◽

Western Blot ◽

Cancer Cells ◽

Cancer Cell ◽

Gastric Cancer Cell ◽

Target Genes ◽

Luciferase Reporter ◽

Gastric Cancer Cells ◽

And Migration

The present study aimed to investigate the effect of miR-3613 on the biological functions of gastric cancer cell lines. The expression of miR-3613 and SOCS4 in gastric cancer cells were detected by RT-qPCR and western blot. The target genes of miR-3613 were verified with the luciferase reporter system and western blot. The SOCS4 overexpression plasmid was constructed and transfected into gastric cancer cells. To further investigate the function of miR-3613, shRNA targeting miR-3613 and SOCS4 overexpression were transfected into SGC-7901. The growth of cells was detected by CCK-8, then the cell invasion and migration ability were detected by wound healing and transwell. Furthermore, the level of cell cycle was detected by flow cytometry. The expression of cell proliferation, cyclin and migration-related proteins were detected by western blot. The results revealed that the expression of miR-3613 is significantly increased in gastric cancer cells. SOCS4 is one of the target genes of miR-3613. Additionally, interference with miR-3613 promotes cell cycle arrest in gastric cancer cells and reversed the inhibitory effect of miR-3613 on biological function of gastric cells. Collectively, the data demonstrated that miR-3613 regulates gastric cancer cell by targeting SOCS4, which is expected to be an attractive target for the development of new drugs for the treatment of gastric cancer.

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Overexpression of NELFE contributes to gastric cancer progression via Wnt/β-catenin signaling-mediated activation of CSNK2B expression

Journal of Experimental & Clinical Cancer Research ◽

10.1186/s13046-021-01848-3 ◽

2021 ◽

Vol 40 (1) ◽

Author(s):

Shijun Yu ◽

Li Li ◽

Hui Cai ◽

Bin He ◽

Yong Gao ◽

...

Keyword(s):

Gastric Cancer ◽

Cell Proliferation ◽

Cancer Progression ◽

Target Genes ◽

Mrna Level ◽

Luciferase Reporter ◽

Migration And Invasion ◽

Loss Of Function ◽

Mice Model ◽

Qrt Pcr

Abstract Background Accumulating evidence has highlighted the importance of negative elongation factor complex member E (NELFE) in tumorigenesis. However, the relationship between NELFE and gastric cancer (GC) remains unclear. This study aimed to explore the expression pattern and specific function of NELFE in GC. Methods NELFE expression was evaluated by immunohistochemistry and qRT-PCR in GC tissues, respectively. Cell proliferation, migration and invasion were measured by CCK-8, colony formation, transwell assays, and nude mice model. Bioinformatics analysis was performed to search potential target genes of NELFE, and a Cignal Finder 10-Pathway Reporter Array was used to explore potential signaling pathways regulated by NELFE. Dual-luciferase reporter assays, qRT-PCR and western blotting were conducted to verify their regulatory relationship. The expression correlations among NELFE, β-catenin and CSNK2B were further explored by immunohistochemistry on consecutive resections. Results NELFE was significantly overexpressed in GC tissues both in protein and mRNA level and negatively correlated with the prognosis of GC patients. Gain- and loss-of-function experiments showed that NELFE potentiated GC cell proliferation and metastasis in vitro and in vivo. CSNK2B was identified as a downstream effector of NELFE. Wnt/β-catenin signaling may mediate the regulation of CSNK2B by NELFE. In addition, NELFE, β-catenin and CSNK2B were all remarkably upregulated in tumor tissues compared with adjacent normal tissues, and their expression levels in GC were positively correlated with each other. Conclusion Our findings reveal a new NELFE-Wnt/β-catenin-CSNK2B axis to promote GC progression and provide new candidate targets against this disease.

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Let-7i-5p enhances cell proliferation, migration and invasion of ccRCC by targeting HABP4

BMC Urology ◽

10.1186/s12894-021-00820-9 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Yujie Liu ◽

Xing Hu ◽

Liang Hu ◽

Changjing Xu ◽

Xuemei Liang

Keyword(s):

Cell Proliferation ◽

Target Genes ◽

The Cancer Genome Atlas ◽

Tumor Promoter ◽

Luciferase Reporter ◽

Migration And Invasion ◽

Poor Overall Survival ◽

Cell Renal Cell Carcinoma ◽

Bioinformatics Analyses ◽

Gene Assay

Abstract Background Clear cell renal cell carcinoma (ccRCC) is one of the best-characterized and most pervasive renal cancers. The present study aimed to explore the effects and potential mechanisms of let-7i-5p in ccRCC cells. Methods Using bioinformatics analyses, we investigated the expression of let-7i-5p in The Cancer Genome Atlas (TCGA) database and predicted biological functions and possible target genes of let-7i-5p in ccRCC cells. Cell proliferation assay, wound healing assay and transwell invasion assay were conducted to characterize the effects of let-7i-5p in ccRCC cells. To verify the interactions between let-7i-5p and HABP4, dual-luciferase reporter assay, quantitative real-time polymerase chain reaction, and western blotting were conducted. Rescue experiments were used to investigate the relationship between let-7i-5p and HABP4. Results TCGA data analysis revealed that ccRCC tissues had significantly increased let-7i-5p expression, which was robustly associated with poor overall survival. Further verification showed that ccRCC cell proliferation, migration and invasion were inhibited by let-7i-5p inhibitor but enhanced by let-7i-5p mimics. Subsequently, HABP4 was predicted to be the target gene of let-7i-5p. TCGA data showed that ccRCC tissues had decreased expression of HABP4 and that HABP4 expression was negatively correlated with let-7i-5p. Further verification showed that downregulation of HABP4 expression promoted cell proliferation, migration and invasion. The dual-luciferase reporter gene assay suggested that the let-7i-5p/HABP4 axis was responsible for the development of ccRCC. Conclusion Our results provide evidence that let-7i-5p functions as a tumor promoter in ccRCC and facilitates cell proliferation, migration and invasion by targeting HABP4. These results clarify the pathogenesis of ccRCC and offer a potential target for its treatment.

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Oxidative stress activates NORAD expression by H3K27ac and promotes oxaliplatin resistance in gastric cancer by enhancing autophagy flux via targeting the miR-433-3p

Cell Death and Disease ◽

10.1038/s41419-020-03368-y ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Jizhao Wang ◽

Yuchen Sun ◽

Xing Zhang ◽

Hui Cai ◽

Cheng Zhang ◽

...

Keyword(s):

Oxidative Stress ◽

Gastric Cancer ◽

Target Genes ◽

Function Analysis ◽

Resistance Index ◽

Luciferase Reporter ◽

Luciferase Reporter Gene ◽

Autophagy Flux ◽

Potential Biomarker ◽

Gene Assay

AbstractOxaliplatin resistance undermines its curative effects on cancer and usually leads to local recurrence. The oxidative stress induced DNA damage repair response is an important mechanism for inducing oxaliplatin resistance by activating autophagy. ELISA is used to detect target genes expression. TMT-based quantitative proteomic analysis was used to investigate the potential mechanisms involved in NORAD interactions based on GO analysis. Transwell assays and apoptosis flow cytometry were used for biological function analysis. CCK-8 was used to calculate IC50 and resistance index (RI) values. Dual-luciferase reporter gene assay, RIP and ChIP assays, and RNA pull-down were used to detect the interaction. Autophagy flux was evaluated using electron microscope and western blotting. Oxidative stress was enhanced by oxaliplatin; and oxaliplatin resistance gastric cancer cell showed lower oxidative stress. TMT labeling showed that NORAD may regulate autophagy flux. NORAD was highly expressed in oxaliplatin-resistant tissues. In vitro experiments indicate that NORAD knockdown decreases the RI (Resistance Index). Oxaliplatin induces oxidative stress and upregulates the expression of NORAD. SGC-7901 shows enhanced oxidative stress than oxaliplatin-resistant cells (SGC-7901-R). NORAD, activated by H3K27ac and CREBBP, enhanced the autophagy flux in SGC-7901-R to suppress the oxidative stress. NORAD binds to miR-433-3p and thereby stabilize the ATG5- ATG12 complex. Our findings illustrate that NORAD, activated by the oxidative stress, can positively regulate ATG5 and ATG12 and enhance the autophagy flux by sponging miR-433-3p. NORAD may be a potential biomarker for predicting oxaliplatin resistance and mediating oxidative stress, and provides therapeutic targets for reversing oxaliplatin resistance.

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Downregulated miR-383-5p contributes to the proliferation and migration of gastric cancer cells and is associated with poor prognosis

PeerJ ◽

10.7717/peerj.7882 ◽

2019 ◽

Vol 7 ◽

pp. e7882 ◽

Cited By ~ 9

Author(s):

Chao Wei ◽

Jian-Jun Gao

Keyword(s):

Gastric Cancer ◽

Target Genes ◽

Enrichment Analysis ◽

Functional Enrichment Analysis ◽

Functional Enrichment ◽

Luciferase Reporter ◽

Reporter Assay ◽

Qrt Pcr ◽

And Migration ◽

Proliferation And Migration

Aim The study aims to identify differentially expressed microRNAs (DEMs) in gastric cancer (GC) and explore the expression, prognosis and downstream regulation role of miR-383-5p in GC. Methods The GC miRNA-Seq and clinical information were downloaded from Firebrowse which stores integrated data sourced from The Cancer Genome Atlas database. The DEMs were identified with limma package in R software at the cut-off criteria of P < 0.05 and |log2 fold change| > 1.0 (|log2FC| > 1.0). The expression of miR-383-5p in GC cell lines and 54 paired GC tissues was measured by quantitative real-time polymerase chain reaction (qRT-PCR). The overall survival curve of miR-383-5p and the association between its expression and clinicopathological features were explored. Wound healing and cell counting kit-8 assays were performed to investigate the capacity of miR-383-5p in cell proliferation and migration. The downstream target genes were predicted by bioinformatics tools (miRDB, TargetScan and starBase). The consensus target genes were selected for gene functional enrichment analysis by FunRich v3.0 software. The luciferase reporter assay was performed to verify the potential targeting sites of miR-383-5p on lactate dehydrogenase A (LDHA). Results A total of 21 down-regulated miRNAs (including miR-383-5p) and 202 up-regulated miRNAs were identified by analyzing GC miRNA-Seq data. Survival analysis found that patients with low miR-383-5p expression had a shorter survival time (median survival time 21.1 months) than those with high expression (46.9 months). The results of qRT-PCR indicated that miR-383-5p was downregulated in GC cell lines and tissues, which was consistent with miRNA-Seq data. The expression of miR-383-5p was significantly associated with tumor size and differentiation grade. Besides, overexpression of miR-383-5p suppressed GC cells proliferation and migration. A total of 49 common target genes of miR-383-5p were obtained by bioinformatics tools and gene functional enrichment analysis showed that these predicted genes participated in PI3K, mTOR, c-MYC, TGF-beta receptor, VEGF/VEGFR and E-cadherin signaling pathways. The data showed that expression of miR-383-5p was negatively correlated with target LDHA (r = −0.203). Luciferase reporter assay suggested that LDHA was a target of miR-383-5p. Conclusion The present study concluded that miR-383-5p was downregulated and may act as a tumor suppressor in GC. Furthermore, its target genes were involved in important signaling pathways. It could be a prognostic biomarker and play a vital role in exploring the molecular mechanism of GC.

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Identification of a Key Competing Endogenous RNA Axis, “COL5A1/miR-137-3p/FSTL1”, Associated With Gastric Cancer Progression by Integrated Bioinformatics Analyses and Experimental Verification

10.21203/rs.3.rs-824329/v1 ◽

2021 ◽

Author(s):

Ming Yang ◽

Zhixing Lu ◽

Liang Li ◽

Min Ma ◽

Fei Long ◽

...

Keyword(s):

Gastric Cancer ◽

Cancer Progression ◽

Target Gene ◽

Target Genes ◽

Differentially Expressed ◽

Luciferase Reporter ◽

Migration And Invasion ◽

Bioinformatics Analyses ◽

Immune Infiltration ◽

Differentially Expressed Mirnas

Abstract Background MicroRNAs (miRNAs) and their target genes have been shown to play an important role in gastric cancer (GC), but this role has not been fully clarified. Therefore, our goal was to find the key miRNA-mRNA regulatory network in GC by combining a variety of bioinformatics and experimental analyses. Methods Differentially expressed miRNAs (DEMs) and genes (DEGs) were screened from TCGA and GEO, respectively. Survival-related differentially expressed miRNAs (SRDEMs) were determined by Cox proportional hazards regression and lasso regression analyses. Differentially expressed target genes (DETGs) of SRDEMs were predicted by TargetScan and miRDB and overlapped with DEGs. We constructed a protein–protein interaction (PPI) network of DETGs and conducted weighted gene coexpression network analysis (WGCNA) to screen the hub genes. Then, qRT–PCR and western blotting were performed to detect the expression level, and a dual-luciferase reporter assay was conducted to verify the binding of miRNA and mRNAs. CCK-8, EdU, wound healing and Transwell assays were conducted to compare the proliferation, migration and invasion abilities of GC cells in the different groups. Results We identified 11 SRDEMs and 233 DETGs, from which we selected miR-137-3p and its target gene COL5A1 for further research because of their key roles in the results of the bioinformatics analyses. Then, we showed that miR-137-3p was significantly downregulated in GC and that overexpression of miR-137-3p suppressed the proliferation, invasion and migration of GC cells by targeting COL5A1. Furthermore, we found that COL5A1 could regulate the expression of FSTL1 and GC progression by sponging miR-137-3p. Finally, bioinformatics analyses showed that FSTL1 might promote GC progression by regulating the immune infiltration of GC. Conclusions miR-137-3p played a tumor-suppressive role in GC, and its target gene COL5A1 could competitively bind miR-137-3p to upregulate the expression of FSTL1, which affects immune infiltration.

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