中国兽医科学2024，Vol.54Issue(6)：718-727,10.DOI:10.16656/j.issn.1673-4696.2024.0084
华支睾吸虫IncRNA EV19对人肝星状细胞纤维化相关基因的调控研究
Clonorchis sinensis lncRNA EV19 regulates genes related to fibrosis in human hepatic stellate cells
摘要
Abstract
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.关键词
华支睾吸虫/人肝星状细胞/肝纤维化/长链非编码RNA/micro RNAKey words
Clonorchis sinensis/human hepatic stellate cells/liver fibrosis/long non-coding RNA/micro RNA分类
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王晶,唐斌,王兴洋,司光泉,刘志娜,丁静,刘明远,刘晓雷,王洋..华支睾吸虫IncRNA EV19对人肝星状细胞纤维化相关基因的调控研究[J].中国兽医科学,2024,54(6):718-727,10.基金项目
中央高校基本科研业务费专项(JLUXKJC2021ZZ10) （JLUXKJC2021ZZ10）
