Modulation of miR-26a-5p and miR-15b-5p Exosomal Expression Associated with Clopidogrel-Induced Hepatotoxicity in HepG2 Cells

Abstract

Clopidogrel is an essential antiplatelet drug used to prevent thrombosis complications associated with atherosclerosis. However, hepatotoxicity is a potential adverse effect related to clopidogrel therapy. Exosome-derived miRNAs may be useful for improved monitoring of drug response and hepatotoxicity risk. In the present study, the expression of several exosomal miRNAs (miR-26a-5p, miR-145-5p, miR-15b-5p, and miR-4701-3p) and cell-derived mRNA targets (PLOD2, SENP5, EIF4G2, HMGA2, STRADB, and TLK1) were evaluated in HepG2 cells treated with clopidogrel (6.25, 12.5, 25, 50, and 100 μM) for 24 and 48 h. Then, clopidogrel cytotoxicity was evaluated by analyzing DNA fragmentation and the cell cycle profile using flow cytometry. Differential expression of exosome-derived miRNAs and cell-derived mRNAs was analyzed by RT-qPCR. Exposure of HepG2 cells to high concentrations of clopidogrel (50 and 100 μM) for 24 h caused significant DNA fragmentation (17.6 and 44.4%, respectively; p < 0.05) and 48 h (26.8 and 48.9%, respectively; p < 0.05), indicating cellular toxicity. Upregulation of miR-26a-5p and downregulation of miR-15b-5p was observed in cells exposed to 100 μM clopidogrel for 24 and 48 h. The miR-26a-5p target mRNAs HMGA2, EIF4G2, STRADB, and SENP5 were downregulated in HepG2 cells following exposure to cytotoxic concentrations of clopidogrel (50 and 100 μM) for 24 h, and HMGA2 levels remained low after 48 h of treatment. TLK1, a target of miR-15b-5p, was downregulated by 50 and 100 μM clopidogrel at 24 h. In conclusion, our results suggest that exposure to high concentrations of clopidogrel modulates the expression of exosomal miR-26a-5p and miR-15b-5p and their target mRNAs in HepG2 cells. Dysregulation of these miRNAs maybe modulate the regulatory pathways involved in clopidogrel-induced liver injury.

SHU00238 Promotes Colorectal Cancer Cell Apoptosis Through miR-4701-3p and miR-4793-3p

“…However, extensive studies are needed to explore the participation of miR-4701-3p and miR-4793-3p in the regulation of SHU00238 on CRC tumor in animals and human (Liang et al, 2017). MiR-4701-3p, derived from exosomes, was shown to influence clopidogrel-induced liver injury, platelet reactivity, and drug-induced toxicity (Freitas et al, 2016;de Freitas et al, 2017). MiR-4701-3p was downregulated in patients with high risk of cadiovascular disease.…”

Construction and Analysis of circRNA-miRNA-mRNA Molecular Regulatory Networks During Herba Gelsemium elegans Intoxication

“…MiR-15a-5p was found to be involved in cell survival and apoptosis (Chen et al, 2017a; Chen and Tian, 2017; Zhou et al, 2018). In addition, dysregulated miR-15b-5p is also related to cellular toxicity (de Freitas et al, 2017; Chen et al, 2018b). Thus, we could conclude that the mmu-circRNA-013703 is involved in GE intoxication by sponging intoxication-related miRNAs.…”

High Mobility Group A (HMGA): Chromatin Nodes Controlled by a Knotty miRNA Network

“…Via the transfection of cells with vectors containing a wild-type or mutated 3 UTR of HMGA1 or HMGA2 and reporter gene assays, several studies have demonstrated that miR-26a specifically targets well-conserved regions of the 3 UTRs of HMGA1 [87,89,90,92,95] and HMGA2 [91,93,94]. miR-26a downregulates HMGA1 at both the mRNA and protein levels, as assessed by qRT-PCR and Western blot analyses [87,90,92], while HMGA2 downregulation has only been validated at the protein level [93,222]. The role of the miR-26a/HMGA1 axis in the context of cancer has been extensively studied, and this axis has been shown to act on the proliferation and migration of pancreatic cancer [96], bladder cancer [95], breast cancer [92], lung adenocarcinoma [90], and osteosarcoma [89] cells.…”