MicroRNA-561 Promotes Acetaminophen-Induced Hepatotoxicity in HepG2 Cells and Primary Human Hepatocytes through Downregulation of the Nuclear Receptor Corepressor Dosage-Sensitive Sex-Reversal Adrenal Hypoplasia Congenital Critical Region on the X Chromosome, Gene 1 (DAX-1)

Abstract

One of the major mechanisms involved in acetaminophen (APAP)-induced hepatotoxicity is hepatocyte nuclear factor 4a (HNF4a)-mediated activation of pregnane X receptor (PXR) and constitutive androstane receptor (CAR). In the present study, we investigated the role of miR-561 and its target gene DAX-1 encoding a corepressor of HNF4a in the process of APAP-induced hepatotoxicity. We used both human hepatocellular liver carcinoma cell line (HepG2) cells and primary human hepatocytes in this study and monitored the levels of reactive oxygen species, lactate dehydrogenase, and glutathione. Our bioinformatics study suggests an association between miR-561 and DAX-1, but not HNF4a. Treatment of HepG2 cells with APAP significantly reduced the expression of DAX-1 in a concentration-dependent manner. miR-561 was induced by APAP treatment in HepG2 cells. Transfection of HepG2 cells with an miR-561 mimic exacerbated APAP-induced hepatotoxicity. HNF4a is physically associated with DAX-1 in HepG2 cells. A decreased protein level of DAX-1 by APAP treatment was also enhanced by miR-561 mimic transfection in HepG2 cells and primary human hepatocytes. The basal and APAP-induced expression of PXR and CAR was enhanced by miR-561 mimic transfection; however, transfection of HepG2 cells or primary human hepatocytes with a miR-561 inhibitor or DAX-1 small interfering RNA reversed these effects. Additionally, the chromatin immunoprecipitation assay revealed that recruitment of DAX-1 onto the PXR promoter was inversely correlated with the recruitment of peroxisome proliferator-activated receptor-a coactivator-1a and HNF4a on APAP treatment. These results indicate that miR-561 worsens APAP-induced hepatotoxicity via inhibition of DAX-1 and consequent transactivation of nuclear receptors.

The miR-561-5p/CX3CL1 Signaling Axis Regulates Pulmonary Metastasis in Hepatocellular Carcinoma Involving CX3CR1+ Natural Killer Cells Infiltration

“…Identification of miR-561-5p was somewhat surprising, as very little is known about its role in cancer 21.To determine the role of miR-561-5p, we speculated that it can modulate cytokine or chemokine levels to reshape the tumor microenvironment to favor metastasis 22. CX 3 CL1 mRNA was identified as the direct downstream target of miR-561-5p, with miR-561-5p overexpression in HCC patients reducing CX 3 CL1 levels, indicating that the crosstalk between cancer cells and their associated stroma cells is potentially regulated by CX 3 CL1.…”

Molecular Biomarkers in Drug-Induced Liver Injury: Challenges and Future Perspectives

“…• Treatment with some immune-related drugs may be effective once the role of immunity mechanism is determined. Tang et al, 2013;Li et al, 2014;Metushi et al, 2014a;He et al, 2015;Metushi et al, 2016 OPN • Prognosis of bad outcome in DILI.…”

“…Specifically, bortezomib was reported to decrease hepatocyte nuclear factor−1a-induced promoter activation of cytochrome P450 2E1 (CYP2E1) and induced endoplasmic reticulum stress, thereby significantly downregulating CYP2E1 expression and alleviating APAP-, CCl 4 -, and thioacetamide-induced liver injury in mice (Park et al, 2016). In addition, certain cytochrome enzymes, including CYP2E1, CYP2B6, and CYP3A4, were associated with the mechanism responsible for causing increased hepatic necrosis (Li et al, 2014). Further, since the occurrence of DILI has been related to genetic vulnerability, many studies have focused on single nucleotide polymorphisms (SNP) in transporter genes (Au et al, 2011).…”

“…Basic studying data GLDH McGill et al, 2012;Harrill et al, 2012;Antoine et al, 2013;Schomaker et al, 2013;McGill et al, 2014;Singhal et al, 2014-K18 Koelink et al, 2009Possamai et al, 2013;Thulin et al, 2014;Xie et al, 2019Cummings et al, 2008SDH Harrill et al, 2012Metushi et al, 2012Metushi et al, 2014bGST Sun et al, 2008Singla et al, 2014;Cai et al, 2015;Wu et al, 2016a;Sun et al, 2017;Yang et al, 2019Giffen et al, 2002Bailey et al, 2012;Tsai et al, 2017BAs James et al, 2015Longo et al, 2019Yamazaki et al, 2013Qiu et al, 2016;Cepa et al, 2018CYP450 Sun et al, 2008Tang et al, 2013;Tang et al, 2013;Metushi et al, 2014a;Li et al, 2014Park et al, 2016OPN Arai et al, 2006Srungaram et al, 2015He et al, 2012Arriazu et al, 2017HMGB1 Antoine et al, 2012…”

DAX-1 Inhibits Hepatocellular Carcinoma Proliferation by Inhibiting β-Catenin Transcriptional Activity

“…Besides, DAX-1 was shown to interact with liver X receptor (LXR) and farnesoid X receptor (FXR) to regulate hepatic triglyceride and bile acid metabolism, respectively [14,15]. A recent study also suggest that DAX-1 participated in the acetaminophen (APAP)-induced hepatotoxicity through regulation of pregnane X receptor (PXR) and constitutive androstane receptor (CAR) [16]. Therefore, the roles and molecular mechanisms of DAX-1 in livers might be diverse and complicated, which remains to be explored in the future.…”