Chemokine receptor CCR6 expression is regulated by miR-518a-5p in colorectal cancer cells

Abstract

BackgroundRecently, involvement of the chemokine/receptor system CCL20/CCR6 in colorectal cancer (CRC) progression was shown. Here, we analyzed the functional interaction of miRNA-518-5p (miR-518a-5p) with CCR6 and its impact on CCR6 expression in CRC cells.MethodsMiR-518a-5p was identified by computer software to potentially interact with CCR6. Hence, functional implications of miR-518a-5p with the 3′UTR of CCR6 were analyzed using the Dual Luciferase Reporter assay system. Confirmation of the predicted target site for miR-518a-5p was achieved by site-directed mutagenesis of the seed sequence in the 3′UTR of CCR6 and subsequent application of the mutated seed sequence in a luciferase assay with miR-518a-5p mimics. Accordingly, two CRC cell lines (Caco-2 and HT-29) were transfected with miR-518a-5p miRNA mimics and gene and protein expression of CCR6 was monitored using qRT PCR and immunocytochemistry, respectively.ResultsAddition of miR-518a-5p led to significant down-regulation of luciferase activity (P < 0.05), which was significantly reversed in a reporter test system containing the mutated seed sequences in the 3′UTR of CCR6. Following transfection of CRC cell lines with miR-518a-5p mimics and subsequent monitoring of CCR6 expression showed significant down-regulation of CCR6 mRNA and CCR6 protein expression in both CRC cell lines under investigation (P < 0.05).ConclusionsWe have shown that miR-518a-5p functionally interacts with CCR6 and that transfection of CRC cells with miR-518a-5p leads to significant CCR6 down-regulation. Consequently, CCR6 expression is regulated by miR-518a-5p in CRC cells indicating that regulation of CCR6 expression by miR-518a-5p might be a regulatory mechanism involved in CRC pathogenesis.

Elevation of circ‐PITX1 upregulates interleukin 17 receptor D expression via sponging miR‐518a‐5p and facilitates cell progression in glioma

“…In this project, we also predicted and verified miR‐518a‐5p as the target of circ‐PITX1. miR‐518a‐5p is a tumor suppressive miRNA in colorectal cancer reported previously . We further identified the tumor suppressor role of miR‐518a‐5p in GM.…”

Circular RNA hsa_circ_0078607 suppresses ovarian cancer progression by regulating miR-518a-5p/Fas signaling pathway

“…To better understand the regulatory mechanism of circ_0078607 in ovarian cancer, we analysed the miRNAs known to be bound by circ_0078607, Bioinformatics analysis identified that miR-518a-5p is a hsa_circ_0078607 associated miRNA and the sponge adsorption effect of circ_0078607 on miR-518a-5p was further verified by dual-luciferase reporter gene assay. Previously, miR-518a-5p has been identified as tumor regulator in many malignancies, including colorectal cancer [23], astrointestinal stromal tumors [24] and germ cell tumors [25]. We found that miR-518a-5p was upregulated in ovarian cance tissues and correctly negatively with circ_0078607 in ovarian cancetissues.…”

An Information Entropy-based Method to Detect microRNA Regulatory Module

“…For example, five of ten miRNAs in MRM 3 are from the same miRNA family MIPF0000020. Furthermore, the five miRNAs play significant roles in varieties of cancers in a coordinated manc 2019 Information Processing Society of Japan ner [25], [26], [27]. Similarly, in MRM 5, three of eleven miR-NAs are from the same miRNA family MIPF0000005, which inhibit or promote tumor growth in a synergistic manner [28], [29].…”