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MiRNA profile in esophageal squamous cell carcinoma: downregulation of miR-143 and miR-145.

ABSTRACT: To investigate the expression profile of miRNA in esophageal squamous cell carcinoma (ESCC).The expression profile of miRNA in ESCC tissues was analyzed by miRNA microarray. The expression levels of miR-143 and miR-145 in 86 ESCC patients were determined by real-time polymerase chain reaction (PCR) using TaqMan assay. The mobility effect was estimated by wound-healing using esophageal carcinoma cells transfected with miRNA expression plasmids.A set of miRNAs was found to be deregulated in the ESCC tissues, and the expression levels of miR-143 and -145 were significantly decreased in most of the ESCC tissues examined. Both miR-143 and miR-145 expression correlated with tumor invasion depth. The transfection of human esophageal carcinoma cells with miR-143 and miR-145 expression plasmids resulted in a greater inhibition of cell mobility, however, the protein level of the previously reported target of miR-145, FSCN1, did not show any significant downregulation.These findings suggest that the deregulation of miRNAs plays an important role in the progression of ESCC. Both miR-143 and miR-145 might act as anti-oncomirs common to ESCC.

SUBMITTER: Wu BL

PROVIDER: S-EPMC3016684 | biostudies-literature | 2011 Jan

REPOSITORIES: biostudies-literature

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MiRNA profile in esophageal squamous cell carcinoma: downregulation of miR-143 and miR-145.

Wu Bing-Li BL Xu Li-Yan LY Du Ze-Peng ZP Liao Lian-Di LD Zhang Hai-Feng HF Huang Qiao Q Fang Guo-Qiang GQ Li En-Min EM

World journal of gastroenterology 20110101 1

<h4>Aim</h4>To investigate the expression profile of miRNA in esophageal squamous cell carcinoma (ESCC).<h4>Methods</h4>The expression profile of miRNA in ESCC tissues was analyzed by miRNA microarray. The expression levels of miR-143 and miR-145 in 86 ESCC patients were determined by real-time polymerase chain reaction (PCR) using TaqMan assay. The mobility effect was estimated by wound-healing using esophageal carcinoma cells transfected with miRNA expression plasmids.<h4>Results</h4>A set of ...[more]

PMID: 21218087

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Project description:MicroRNAs (miRNAs), 18-24 nt non-coding RNAs, are thought to play important roles in cell proliferation, differentiation, apoptosis, and development. Recent studies suggest that some of the known microRNAs map to a single genomic locale within a single polycistronic transcript. But the roles of the cluster remain to be known. In order to understand the role and mechanism of a cluster of miR-143 and miR-145 in esophageal squamous cell carcinoma (ESCC), the association of mature miR-143 and miR-145 expression with the risk for esophageal cancer was evaluated in ESCC patients with a case-control study, and target protein regulated by mature miRNA was analyzed in ESCC cell lines with 3'UTR luciferase reporter assay. The expression levels of miR-143 and miR-145 were determined in 110 pairs of esophageal cancer tissues and adjacent normal tissues using real-time reverse transcription PCR. The relative expression of miR-143 and miR-145 were statistically different between cancer tissues and matched controls. The combined expression of miR-143 and miR-145 was significantly associated with the risk for esophageal cancer. Meanwhile, the reduced expression of two miRNAs in tumor patient was supposed to have a trend of lymph node metastases. The co-expression pattern of miR-143 and miR-145 was analyzed with Pearson correlation. It showed a significant correlation between these two miRNAs expression both in tissues and tumor cell lines. 3'UTR luciferase reporter assay indicated that Fascin Homolog 1 (FSCN1) could be co-regulated by miR-143 and miR-145. The protein level of FSCN1 showed no significant linear correlation with miR-143 and miR-145 expression in ESCC cell lines with Western blotting analysis. In conclusion, since miR-143 and miR-145 could regulate oncogenic FSCN1 and take part in the modulation of metastases, the result suggested the combination variable of miR-143 and miR-145 as a potential biomarker for earlier diagnosis and prognosis of esophageal cancer.

Project description:BackgroundThe impact of miR-145-5p in immune infiltration and the potential application in esophageal squamous cell carcinoma (ESCC) immunochemotherapy remains unknown.MethodsTranscriptomic data for ESCC tissues and normal tissues and clinical materials of patients with ESCC were obtained from The Cancer Genome Atlas (TCGA) and the Gene Expression Omnibus (GEO) databases. The differences in mRNA levels in cancer tissues and noncancerous tissues were analyzed, and we subsequently investigated the association between miR-145-5p expression and the key parameters of ESCC progression and prognosis. Additionally, cytological experiments were performed to evaluate the biological functions of miR-145-5p. Pathways potentially affected by miR-145-5p were analyzed by Gene Set Enrichment Analysis (GSEA) and REACTOME. We also analyzed the function of miR-145-5p in immune infiltration through the TIMER2 (Tumor Immune Estimation Resource) database.ResultsThe analysis of gene chip data from the TCGA database and GEO database (including GSE13937 and GSE43732) showed that the expression of miR-145-5p is downregulated in ESCC (P<0.05) and that patients with high miR-145-5p levels had lower survival rates (P<0.05). The expression of miR-145-5p was significantly correlated with the disease-free survival (DFS) rate (P<0.05) and M stage (P<0.05) in the TCGA database and was significantly correlated with the T stage (P<0.05) and TNM stage (P<0.05) in the GSE13937 database. Functional experiments showed that miR-145-5p attenuated proliferation (P<0.05), migration (P<0.01) and invasion (P<0.01) in the Eca109 cell line. Both GSEA gene enrichment and REACTOME gene enrichment revealed that miR-145-5p was associated with tumor signaling pathways and immune signaling pathways. Immune infiltration analysis revealed that the expression level of miR-145-5p was significantly correlated with the infiltration level of macrophages (P<0.05) and was positively correlated with the level of gene markers of M2 macrophages and tumor-associated macrophages (P<0.05).ConclusionsMiR-145-5p acts as a tumor suppressor microRNA in ESCC and is an important noncoding RNA in the high M2-like tumor-associated macrophage infiltration of ESCC. Assessing the miR-145-5p level in ESCC samples has translational meaning, which help illustrate the immune infiltration status, predict the prognostic outcome, and select the type of immunochemotherapy.

Project description:MicroRNAs (miRNAs) are an endogenous conserved class of non-coding 20–22 nt small RNAs that regulate gene expression at post-transcriptional level by mostly binding to 3′-UTR of target mRNAs, leading to mRNA degradation or translation inhibition. Recent reports demonstrate a role for miRNA expression in the disease progression and outcome. By now, many researcher focusing on miRNA expression profiles in Barrett's esophagus and esophageal adenocarcinoma have been reported. Nevertheless, there is still a little information available about specific miRNA expression pattern and their roles in ESCC. To develop novel diagnostic and therapeutic targets for esophageal squamous cancer, we first investigated the expression profile of miRNA in three pairs of ESCC clinical samples. Tissues of ESCC and the matched normal counterparts were obtained from surgical specimens immediately after resection from patients undergoing primary surgical treatment of esophageal carcinoma from the Department of Tumor Surgery of Shantou Central Hospital, China. RNA labeling and hybridization were completed by KangChen Bio-tech Inc. (Shanghai, China) according to the manufacturer's instructions. Briefly, total RNA from three pairs of esophageal carcinoma and matched normal tissues were isolated by Trizol (Invitrogen, USA) and purified by RNeasy mini kit (QIAGEN, German). The concentration and quality of total RNA were measured by NanoDrop ND-1000 at 260 and 280 nm (A260/280) and checked by gel electrophoresis. Each RNA sample from three pairs of ESCC was separately labeled either using the miRCURY Hy3/Hy5 labeling kit and hybridized on the six miRCURYTM locked nucleic acid (LNA) array version 11.0 (Exiqon, Denmark), which contains probes for 1700 mature miRNA. Scans were quantified by using GenePix software (Molecular Devices). The data were exported to Microsoft Excel worksheets, log2 transformed, normalized using global Lowess (Locally Weighted Scatter plot Smoothing) regression algorithm (MIDAS, TIGR Microarray Data Analysis System).

Dataset Information

MiRNA profile in esophageal squamous cell carcinoma: downregulation of miR-143 and miR-145.

Publications

MiRNA profile in esophageal squamous cell carcinoma: downregulation of miR-143 and miR-145.

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