Yan Xin, Li Qianyin, Ni Dongsheng, Xie Yajun, He Qingling, Wan Qianya, Liu Yamin, Lyu Zhongshi, Mao Zhaomin, Zhou Qin
1 The College of Laboratory Medicine, Chongqing Medical University, Chongqing, China.
2 Division of Molecular Nephrology and the Creative Training Center for Undergraduates, The Ministry of Education Key Laboratory of Laboratory Medical Diagnostics, The College of Laboratory Medicine, Chongqing Medical University, Chongqing, China.
Tumour Biol. 2017 Jun;39(6):1010428317706218. doi: 10.1177/1010428317706218.
A1CF (apobec-1 complementation factor) acts as a component of the apolipoprotein-B messenger RNA editing complex. Previous researches mainly focused on its post-transcriptional cytidine to uridine RNA editing. However, few study reported its role in progression of breast carcinoma cells. Wound healing assay and flow cytometry were applied to detect the migration and apoptosis; western blot, real-time polymerase chain reaction, and dual-luciferase assays were applied to investigate the potential regulation mechanism of A1CF-mediated cell migration and apoptosis. Knockdown of A1CF decreased cell migration and enhanced cell apoptosis in MCF7 cells in vitro. Western blot analysis showed that knockdown of A1CF decreased Dickkopf1 but increased c-Myc and β-catenin expression, and overexpression of A1CF can get opposite results. Knockdown of Dickkopf1 in A1CF-overexpressed cells decreased cell migration and enhanced cell apoptosis compared with A1CF-overexpressed cells. Luciferase-fused 3' untranslated region of human Dickkopf1 activity was highly upregulated in A1CF-overexpressed MCF7 cells, but this upregulation can be inhibited by mutating conserved binding motifs of Dickkopf1 3' untranslated region. A1CF played a crucial role in cell migration and survival through affecting 3' untranslated region of Dickkopf1 to upregulate its expression in MCF7 cells.
A1CF（载脂蛋白B信使核糖核酸编辑复合体互补因子）作为载脂蛋白B信使核糖核酸编辑复合体的一个组成部分发挥作用。以往的研究主要集中在其转录后胞嘧啶到尿嘧啶的核糖核酸编辑。然而，鲜有研究报道其在乳腺癌细胞进展中的作用。采用伤口愈合试验和流式细胞术检测细胞迁移和凋亡；采用蛋白质免疫印迹法、实时聚合酶链反应和双荧光素酶测定法研究A1CF介导的细胞迁移和凋亡的潜在调控机制。在体外，敲低A1CF可降低MCF7细胞的迁移并增强细胞凋亡。蛋白质免疫印迹分析表明，敲低A1CF可降低Dickkopf1的表达，但增加c-Myc和β-连环蛋白的表达，而A1CF的过表达则会得到相反的结果。与A1CF过表达的细胞相比，在A1CF过表达的细胞中敲低Dickkopf1可降低细胞迁移并增强细胞凋亡。在A1CF过表达的MCF7细胞中，荧光素酶融合的人Dickkopf1 3'非翻译区活性高度上调，但这种上调可通过突变Dickkopf1 3'非翻译区的保守结合基序来抑制。在MCF7细胞中，A1CF通过影响Dickkopf1的3'非翻译区来上调其表达，从而在细胞迁移和存活中发挥关键作用。
