Department of Pediatric Hepatobiliary Surgery, Children's Hospital of Fudan University, Shanghai 201102, P.R. China.
Int J Mol Med. 2019 Feb;43(2):739-748. doi: 10.3892/ijmm.2018.4023. Epub 2018 Dec 11.
Biliary atresia (BA) is the most common cause of chronic cholestasis in children. The long non‑coding RNA (lncRNA) Annexin A2 pseudogene 3 (ANXA2P3) and Annexin A2 (ANXA2) have been suggested to serve pivotal roles in BA; however, the clinical significance and biological roles of ANXA2P3 and ANXA2 in BA remain to be elucidated. The present study aimed to elucidate the function of ANAX2P3 and ANXA2 in BA‑induced liver injury using a human liver cell line and liver tissues from patients with BA. Reverse transcription‑quantitative polymerase chain reaction, western blotting and immunohistochemistry were conducted to determine the expression levels of ANXA2 and ANXA2P3 in liver tissues from patients with BA. Classification of fibrosis was analyzed by Masson staining. The functional roles of ANXA2 and ANXA2P3 in liver cells were determined by Cell Counting kit‑8 assay, and flow cytometric and cell cycle analyses. Activation of the ANXA2/ANXA2P3 signaling pathway in liver cells was evaluated by western blot analysis. According to the present results, the expression levels of ANXA2 and ANXA2P3 were significantly increased in liver tissues from patients with BA. In addition, knocking down the expression of ANXA2P3 and ANXA2 may result in reduced liver cell proliferation, cell cycle arrest in G1 phase and increased apoptosis of liver cells in vitro. Furthermore, in cells in which ANXA2 and ANXA2P3 were overexpressed, cell apoptosis was reduced and cell cycle arrest in G2 phase. Taken together, these results indicated that ANXA2P3 and ANXA2 may have protective effects against liver injury progression and may be considered biomarkers in patients with BA.
先天性胆道闭锁（BA）是儿童慢性胆汁淤积的最常见原因。长链非编码 RNA（lncRNA）膜联蛋白 A2 假基因 3（ANXA2P3）和膜联蛋白 A2（ANXA2）被认为在 BA 中发挥关键作用；然而，ANXA2P3 和 ANXA2 在 BA 中的临床意义和生物学作用仍有待阐明。本研究旨在通过人肝细胞系和 BA 患者的肝组织阐明 ANXA2P3 和 ANXA2 在 BA 诱导的肝损伤中的作用。采用逆转录定量聚合酶链反应、western blot 及免疫组织化学法检测 BA 患者肝组织中 ANXA2 和 ANXA2P3 的表达水平。采用 Masson 染色法分析纤维化分级。通过细胞计数试剂盒-8 检测、流式细胞术和细胞周期分析确定 ANXA2 和 ANXA2P3 在肝细胞中的功能作用。通过 western blot 分析评估 ANXA2/ANXA2P3 信号通路在肝细胞中的激活情况。根据本研究结果，BA 患者肝组织中 ANXA2 和 ANXA2P3 的表达水平显著升高。此外，敲低 ANXA2P3 和 ANXA2 的表达可能导致体外肝细胞增殖减少、G1 期细胞周期停滞和细胞凋亡增加。此外，在过表达 ANXA2 和 ANXA2P3 的细胞中，细胞凋亡减少，G2 期细胞周期停滞。综上所述，这些结果表明 ANXA2P3 和 ANXA2 可能对肝损伤进展具有保护作用，可作为 BA 患者的生物标志物。
