Department of General Surgery, Shenzhen Children's Hospital, Shenzhen, China.
Department of Pediatric Surgery, Shantou University Medical College, Shantou, China.
Bioengineered. 2022 Mar;13(3):6222-6230. doi: 10.1080/21655979.2022.2041321.
Biliary atresia (BA) is a devastating liver disease in neonates. Liver fibrosis is regarded as a universal and prominent feature of BA. Studies have revealed that long non-coding RNAs (lncRNAs) regulate cellular processes during the development of liver fibrosis in BA. Long non-coding RNA-adducin 3 antisense RNA1 (lnc-ADD3-AS1) has been shown to increase susceptibility to BA. However, the role of lnc-ADD3-AS1 in liver fibrosis in BA remains unclear. Here, we investigated the role of lnc-ADD3-AS1 in the proliferation, migration, and apoptosis of the immortalized human hepatic stellate cell (HSC) line, LX-2. We successfully overexpressed and silenced lnc-ADD3-AS1 in LX-2 cells using adenovirus vectors and evaluated the proliferation of transfected cells using the Cell Counting Kit-8 (CCK8) assay. Cell apoptosis was detected using annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) double staining and flow cytometry. We then analyzed cell migration by performing wound-scratch and transwell migration assays. Our results show that lnc-ADD3-AS1 significantly promoted LX-2 cell proliferation and attenuated apoptosis. More importantly, lncRNA-ADD3-AS1 significantly accelerated the migration of LX-2 cells. Our data indicated that lncRNA-ADD3-AS1 plays a role in the pathogenesis of liver fibrosis in patients with BA and may serve as a potential diagnostic marker for monitoring liver fibrosis in BA or as a therapeutic target for the disease.
先天性胆道闭锁(BA)是一种新生儿期发生的毁灭性肝脏疾病。肝纤维化被认为是 BA 的普遍而突出的特征。研究表明,长链非编码 RNA(lncRNA)在 BA 肝纤维化的发展过程中调节细胞过程。长链非编码 RNA-黏附素 3 反义 RNA1(lnc-ADD3-AS1)已被证明增加了 BA 的易感性。然而,lnc-ADD3-AS1 在 BA 肝纤维化中的作用尚不清楚。在这里,我们研究了 lnc-ADD3-AS1 在永生化人肝星状细胞(LX-2)系增殖、迁移和凋亡中的作用。我们成功地使用腺病毒载体过表达和沉默 LX-2 细胞中的 lnc-ADD3-AS1,并使用细胞计数试剂盒-8(CCK8)测定法评估转染细胞的增殖。通过 Annexin V-荧光素异硫氰酸酯(FITC)/碘化丙啶(PI)双重染色和流式细胞术检测细胞凋亡。然后,我们通过划痕和 Transwell 迁移实验分析细胞迁移。我们的结果表明,lnc-ADD3-AS1 显著促进了 LX-2 细胞的增殖并减弱了细胞凋亡。更重要的是,lncRNA-ADD3-AS1 显著加速了 LX-2 细胞的迁移。我们的数据表明,lncRNA-ADD3-AS1 在 BA 患者肝纤维化的发病机制中起作用,并且可能作为监测 BA 肝纤维化的潜在诊断标志物或作为该疾病的治疗靶点。