Effects of Ethanol on Expression of Coding and Noncoding RNAs in Murine Neuroblastoma Neuro2a Cells

Abstract

Excessive use of alcohol can induce neurobiological and neuropathological alterations in the brain, including the hippocampus and forebrain, through changes in neurotransmitter systems, hormonal systems, and neuroimmune processes. We aimed to investigate the effects of ethanol on the expression of coding and noncoding RNAs in a brain-derived cell line exposed to ethanol. After exposing Neuro2a cells, a neuroblastoma cell line, to ethanol for 24 and 72 h, we observed cell proliferation and analyzed up- and downregulated mRNAs and long noncoding RNAs (lncRNAs) using total RNA-Seq technology. We validated the differential expression of some mRNAs and lncRNAs by RT-qPCR and analyzed the expression of Cebpd and Rnu3a through knock-down of Cebpd. Cell proliferation was significantly reduced in cells exposed to 100 mM ethanol for 72 h, with 1773 transcripts up- or downregulated by greater than three-fold in ethanol-treated cells compared to controls. Of these, 514 were identified as lncRNAs. Differentially expressed mRNAs and lncRNAs were mainly observed in cells exposed to ethanol for 72 h, in which Atm and Cnr1 decreased, but Trib3, Cebpd, and Spdef increased. On the other hand, lncRNAs Kcnq1ot1, Tug1, and Xist were changed by ethanol, and Rnu3a in particular was greatly increased by chronic ethanol treatment through inhibition of Cebpd. Our results increase the understanding of cellular and molecular mechanisms related to coding and noncoding RNAs in an in vitro model of acute and chronic exposure to ethanol.

Prospective DNA Methylation Analysis of the CpG GABRA2 Receptor Subunit in Alcohol Dependence during Detoxification

“…This is higher than the average blood alcohol concentration in alcoholdependent subjects. Moreover, it has been reported that in vitro alcohol treatment using concentrations above 100 mmol/L has a direct cytotoxic effect of alcohol on cells [28]. On the other hand, several recent studies could not confirm this observation [29][30][31].…”

Ethanol responsive lnc171 promotes migration and invasion of HCC cells via mir-873-5p/ZEB1 axis

“…1 d). Some studies have reported that ethanol could induce changes of lncRNA levels in cells [ 17 – 19 ], so we constructed a cell model of ethanol treatment to observe whether ethanol can cause changes of lnc171 levels in HCC cells. Before establishing the cell model, we treated cells with gradient concentration of ethanol and measured cellular activity at 24 h, 48 h, and 72 h after ethanol treatment.…”

“…Ethanol can induce liver damage and hepatic steatosis these lesions can cause alcoholic liver disease to progress to cirrhosis and eventually to HCC [ 35 ]. Ethanol has been proved to be a teratogenic agent and epigenetic regulator that regulates cell biological traits and functions by regulating the expression levels of multiple factors, leading to aggravating liver damage and accelerating the process of liver lesions [ 17 – 19 ]. In our study, we found that the stimulation of ethanol could increase the expression of lnc171 in HCC cells, and the migration and invasion ability of HCC cells was enhanced.…”

“…For example, alcohol, one of the risk factors of HCC, that could induce alcoholic hepatitis which gradually develop into alcoholic cirrhosis and eventually leading to the occurrence of HCC [ 16 ]. And alcohol have been shown to be an epigenetic modulator that could stimulate cells to secreted variety of cytokine factors including lncRNA [ 17 – 19 ]. However, the function and mechanism of lncRNA in the process of alcohol-induce liver injury and HCC remains unknow.…”