Clonorchis sinensis lncRNA EV19 regulates genes related to fibrosis in human hepatic stellate cells
In order to investigate the impact of Clonorchis sinensis long non-coding RNAs(lncRNAs)on human hepatic stellate cells(LX-2 cells)fibrosis-related factors,eight lncRNA of C.sinensis pos-sibly related to fibrosis were initially screened through bioinformatics analysis.And RT-qPCR con-firmed the expression of the lncRNAs EV1,EV17,and EV19 in adult C.sinensis and their extracellular vesicles.Then,pcDNA3.1-lncRNA EV19,pcDNA3.1-lncRNA EV17,and pcDNA3.1-lncRNA EV1 overexpression vectors were constructed,and transfected into LX-2 cells.The transfected cells were collected,and the levels of transcription and protein expression of the α-SMA,Col 1,and Col 3 genes were detected using RT-qPCR and Western-blot,respectively.Additionally,the CCK-8 assay was used to detect the influence of lncRNA on the proliferation of LX-2 cells.Finally,RT-qPCR and a double luciferase reporter assay were performed to verify the target of lncRNA EV19.The results showed that,compared with the control group,the overexpression of lncRNA EV19 significantly increased the transcription and protein levels of α-SMA,Col 1,and Col 3(P＜0.05),and promoted the proliferat ion of LX-2 cells.The RT-qPCR results showed that the expression levels of miR-19b-1-5p,miR-29b-1-5p,and miR-940 were decreased in LX-2 cells overexpressing lncRNA EV19.Dual-luciferase reporter assay confirmed that lncRNA EV19 targets miR-19b-1-5p.These results indicate that the lncRNA EV19 from C.sinensis may promote the proliferat ion of LX-2 cells and induce fibrosis by negatively regulating miR-19b-1-5p.
国家科技期刊平台
NSTL
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