Integrated Analysis of lncRNA-miRNA-mRNA Associated ceRNA Networks Involved in Cystic Echinococcosis-Induced Fibrogenesis in Vitro
JOURNAL OF BIOLOGICAL REGULATORS AND HOMEOSTATIC AGENTS(2023)
Abstract
Background: The primary pathological features of cystic echinococcosis (CE) are the formation of fibrotic pericytes and spaceoccupying lesions in the liver, but fibrogenesis has received little attention in CE. This study aimed to investigate the possible mechanisms underlying CE-induced fibrogenesis from the perspective of competing endogenous RNAs (ceRNAs). Methods: Hydatid fluid was collected from sheep livers infected with Echinococcus granulosus and was used to treat rat hepatic stellate cells (HSC-T6) to establish a fibrotic cell model. Next, RNA sequencing and bioinformatic analyses were conducted. Finally, two important ceRNA axes were chosen for quantitative real-time polymerase chain reaction (qRT-PCR) analysis. Results: After hydatid fluid treatment, the both mRNA and protein expression levels of fibrosis-related factors (anti-alphasmooth muscle actin, type III collagen, and transforming growth factor-beta receptor II) were significantly higher than those in control cells. After sequencing, 569 differentially expressed (DE) mRNAs, 151 DE long non-coding RNAs (lncRNAs), and 14 DE miRNAs were identified in infected cells. Next, ceRNA regulatory networks consisting of 207 mRNAs, 13 miRNAs, and 54 lncRNAs were constructed. Based on the comparative toxicogenomics database, 8 fibrosis-related pathways were identified, including focal adhesion, extracellular matrix-receptor interaction, the hypoxia-inducible factor 1 signaling pathway, and pathways in cancer, as well as disease pathway-related regulatory networks associated with 38 DE lncRNAs, 43 DE mRNAs, 11 DE miRNAs, and 8 pathways. The qRT-PCR results of LNC_003335-novel_559-TNC and LNC_002273-miR-125-2-3p-PDGFD were consistent with the RNA sequencing results. Conclusions: Our study showed that the two ceRNA axes LNC_003335-novel_559-TNC and LNC_002273-miR-125-2-3p-PDGFD may be potential critical targets for CE-induced fibrogenesis.
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Key words
cystic echinococcosis, fibrosis, pathway, competing endogenous RNA, RNA sequencing
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