LOC102553417 silencing facilitates the apoptosis of hepatic stellate cells via the miR-30e/MTDH axis
LOC102553417 silencing facilitates the apoptosis of hepatic stellate cells via the miR-30e/MTDH axis
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DOI：
10.3892/mmr.2022.12865
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发表时间：
2022-09
影响因子：
3.4
通讯作者：
Wujun Wei;Cheng Lin;Rentong Hu;Jingjing Huang;Xiaohao Chen;Lv Zhou;J. Wei;Yi-bin Deng;Chun-Fang Wang
中科院分区：
文献类型：
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作者：
Wujun Wei;Cheng Lin;Rentong Hu;Jingjing Huang;Xiaohao Chen;Lv Zhou;J. Wei;Yi-bin Deng;Chun-Fang Wang
肝纤维化是多种慢性肝病发展过程中不可避免的病理过程，也是肝病治疗的一大挑战。本研究的目的是证明长非编码RNA LOC 102553417的沉默是否通过microRNA（miR）-30 e/metadherin（MTDH）轴促进肝星状细胞（HSC）凋亡。构建L0 C102553417沉默慢病毒并将其转导到HSC-T6细胞中。在通过逆转录-定量PCR确认沉默效率后，使用Cell Counting Kit-8测定法评估细胞增殖，并使用流式细胞术评估细胞凋亡。使用双荧光素酶报告基因测定和RNA结合蛋白免疫沉淀证明了LOC 102553417与miR-30 e之间以及miR-30 e与MTDH之间的相互作用。转染miR-30 e模拟物和抑制剂后，检测是否存在沉默LOC 102553417，检测HSC-T6细胞的凋亡。LOC 102553417基因沉默抑制HSC-T6细胞增殖并促进其凋亡。已证明L0 C102553417靶向miR-30 e，而miR-30 e靶向MTDH。此外，LOC 102553417沉默显著上调miR-30 e表达水平，并显著下调MTDH mRNA和蛋白表达水平，导致p-Akt/Akt比值显著降低，p53蛋白表达水平显著升高。单独转染miR-30 e模拟物可显著促进HSC-T6细胞凋亡，抑制LOC 102553417和MTDH的表达。此外，miR-30 e模拟物可加速LOC 102553417沉默刺激的HSC-T6细胞凋亡;通过对miR-30 e抑制剂的反向验证获得了一致的结果。总之，本研究表明，LOC 102553417沉默通过miR-30 e/MTDH轴刺激HSC凋亡。
Hepatic fibrosis is an inevitable pathological process in the progression of multiple chronic liver diseases and remains a major challenge in the treatment of liver diseases. The purpose of the present study was to demonstrate whether silencing of the long non-coding RNA LOC102553417 promoted hepatic stellate cell (HSC) apoptosis via the microRNA (miR)-30e/metadherin (MTDH) axis. A LOC102553417 silencing lentivirus was constructed and transduced into HSC-T6 cells. After confirming the silencing efficiency by reverse transcription-quantitative PCR, cell proliferation was assessed using the Cell Counting Kit-8 assay and apoptosis was assessed using flow cytometry. The interaction between LOC102553417 and miR-30e, and that between miR-30e and MTDH, was demonstrated using the dual-luciferase reporter assay and RNA binding protein immunoprecipitation. The apoptosis of HSC-T6 cells was detected after transfection of miR-30e mimics and inhibitors with or without silencing LOC102553417. Silencing of LOC102553417 curbed HSC-T6 cell proliferation and expedited their apoptosis. LOC102553417 was demonstrated to target miR-30e, whereas miR-30e targeted MTDH. In addition, LOC102553417 silencing significantly upregulated miR-30e expression levels, and significantly downregulated MTDH mRNA and protein expression levels, which resulted in a significantly reduced p-Akt/Akt ratio and significantly elevated p53 protein expression levels. Transfection with miR-30e mimic alone significantly enhanced HSC-T6 cell apoptosis and inhibits LOC102553417 and MTDH expressions, In addition, miR-30e mimic expedites the apoptosis of HSCs stimulated by LOC102553417 silencing; consistent results were obtained by reverse validation of miR-30e inhibitor. In conclusion, the present study demonstrated that LOC102553417 silencing stimulated the apoptosis of HSCs via the miR-30e/MTDH axis.