key: cord-1019983-1nb1qwjk authors: Shen, Xuehuai; Yin, Lei; Pan, Xiaocheng; Zhao, Ruihong; Zhang, Danjun title: Porcine epidemic diarrhea virus infection blocks Cell cycle and induces apoptosis in pig intestinal epithelial cells date: 2020-07-10 journal: Microb Pathog DOI: 10.1016/j.micpath.2020.104378 sha: 28ab35e89b649aab06d78919fb136cbd1e86dc52 doc_id: 1019983 cord_uid: 1nb1qwjk Porcine epidemic diarrhea virus (PEDV) is responsible for the acute infectious swine disease porcine epidemic diarrhea (PED). PED causes damage to the intestine, including villus atrophy and shedding, leading to serious economic losses to the pig industry worldwide. We carried out an in vitro study to investigate cell apoptosis and the cell cycle in a PEDV-infected host using transcriptomic shotgun sequencing (RNA-Seq) to study gene responses to PEDV infection. Results revealed that the PEDV infection reduced proliferation activity, blocked the cell cycle at S-phase and induced apoptosis in IPEC-J2 cells. The expression of gene levels related to ribosome proteins and oxidative phosphorylation were significantly up-regulated post-PEDV infection. Although the significantly down-regulated on PI3K/Akt signaling pathway post-PEDV infection, the regulator–related genes of mTOR signaling pathway exerted significantly up-regulated or down-regulated in IPEC-J2 cells. These results indicated that ribosome proteins and oxidative phosphorylation process were widely involved in the pathological changes and regulation of host cells caused by PEDV infection, and PI3K/AKT and mTOR signaling pathways played a vital role in antiviral regulation in IPEC-J2 cells. These data might provide new insights into the specific pathogenesis of PEDV infection and pave the way for the development of effective therapeutic strategies. showed that the JNK and p38 mitogen-activated protein kinase (MAPK) pathways also contributed to 68 PEDV infection, while another study showed that activated p53 and the accumulation of reactive 69 oxygen species participated in PEDV-induced apoptosis, and that p53 could be regulated by reactive 70 oxygen species during PEDV infection [11] . Activated p38 MAPK and SAPK/JNK had no effect on Although the understanding of the molecular virology, epidemiology, and vaccinology of PEDV 77 has improved greatly since the reemergence of these strains, the molecular mechanism of the host cell In the present study, we determined the effect of PEDV infection on apoptosis and the cell cycle in (log2-fold change; P < 0.05, false discovery rate > 95%) were selected for real-time qPCR.The primer 168 pairs for qPCR were designed using the assembled RNA-Seq sequence data (Table 1) . qPCR was 169 carried out with a SYBR Green PCR Master Mix Kit (Toyobo, Osaka, Japan) using an ABI7500 Statistical analyses were carried out using Prism software and expressed as the mean ± standard 177 deviation. Differences among multiple groups were analyzed by one-way analysis of variance followed 178 by the Student-Newman-Keuls test. Differences were considered significant at P < 0.05. There 25880 DEGs were normalized and 251 significant DEGs were detected between the 216 PEDV-infected and control groups, including 133 up-regulated and 118 down-regulated genes (Fig. 217 4) . GO functional cluster analysis divided the DEGs into three categories involving biological 218 process, cellular component and molecular function (Fig. 5) . Three categories include 20 219 sub-categories and 55 term. There are the most gene related to cellular process and single-organism process in biological process category. Cell and cell part -related genes are most 221 in cellular component category, and the most genes related to binding and catalytically activity in 222 molecular function. KEGG pathway analysis was performed to evaluate the biological and 223 ontological significance of DEGs. The results showed that the DEGs were involved in a variety of 224 life activities, and more genes were annotated into ribosome, oxidative phosphorylation and signal 225 transduction of viral infection (Fig. 6) . The ribosome is a large assembly of proteins and ribosomal 226 RNAs (rRNAs) that functions to translate messenger RNAs (mRNAs) into proteins. In our results, 227 Table 2 . 251 Thirteen of the significantly differentially expressed genes between the PEDV infection and 253 control groups were selected for real-time qPCR. β-actin as a reference gene for qPCR. The results 254 showed that the expression level of thirteen genes were consistent with the results from 255 RNA-Seq (Fig. 7) . Apoptosis is an important process regulating the pathogenesis of many infectious diseases [10] . 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The authors declared that they have no conflicts of interest to this work.