Integrated MicroRNA–mRNA Analysis Reveals miR-204 Inhibits Cell Proliferation in Gastric Cancer by Targeting CKS1B, CXCL1 and GPRC5A

Abstract

Gastric cancer (GC) is the second most frequent cause of cancer-related deaths worldwide. MicroRNAs are single-stranded RNA molecules of 21–23 nucleotides that regulate target gene expression through specific base-pairing interactions between miRNA and untranslated regions of targeted mRNAs. In this study, we generated a multistep approach for the integrated analysis of miRNA and mRNA expression. First, both miRNA and mRNA expression profiling datasets in gastric cancer from the Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) identified 79 and 1042 differentially expressed miRNAs and mRNAs, respectively, in gastric cancer. Second, inverse correlations between miRNA and mRNA expression levels identified 3206 miRNA–mRNA pairs combined with 79 dysregulated miRNAs and their 774 target mRNAs predicted by three prediction tools, miRanda, PITA, and RNAhybrid. Additionally, miR-204, which was found to be down-regulated in gastric cancer, was ectopically over-expressed in the AGS gastric cancer cell line and all down-regulated targets were identified by RNA sequencing (RNA-seq) analysis. Over-expression of miR-204 reduced the gastric cancer cell proliferation and suppressed the expression of three targets which were validated by qRT-PCR and luciferase assays. For the first time, we identified that CKS1B, CXCL1, and GPRC5A are putative targets of miR-204 and elucidated that miR-204 acted as potential tumor suppressor and, therefore, are useful as a promising therapeutic target for gastric cancer.

CKS1B as Drug Resistance-Inducing Gene—A Potential Target to Improve Cancer Therapy

“…In addition, experiments have shown that miR-1258 exerts a tumor-suppressive effect through immediately downregulation of the oncogenic CKS1B gene in CRC (24). Other studies have shown that miR-204 negatively regulates the expression of CKS1B in gastric cancer (53). Yu Fujita found that miR-197 was downregulated in platinum-resistant NSCLC specimens, leading to in vitro and in vivo chemical resistance, tumorigenicity, and increased lung metastasis.…”

GPRC5A: An Emerging Biomarker in Human Cancer

“…miR-103a-3p has two target sites in the 5'UTR of GPRC5A , and in vitro studies have found that it suppresses the expression of GPRC5A mRNA and protein by binding to either of them [ 35 ]. Besides, miR-204 can inhibit GPRC5A expression via binding to its 3' UTR in gastric cancer (GC) [ 36 ].…”

MiR-204 acts as a potential therapeutic target in acute myeloid leukemia by increasing BIRC6-mediated apoptosis

“…MiR-204 is reported to play a relevant role in the development of gastric cancer ( 8 ), prostate cancer ( 9 , 10 ), ovarian cancer ( 11 ), colorectal cancer ( 12 ), cervical cancer ( 13 ), and melanoma ( 14 ). For example, miR-204 suppressed cell proliferation in gastric cancer by targeting CSK1B, GPRC5A and CXCL1 ( 8 ). Moreover, miR-204 was downregulated in cervical cancer tissues which plays an important role in regulating cervical cancer cell proliferation, migration and invasion ( 13 ).…”