Department of Gastroenterology and Hepatology, Erasmus Medical Center Cancer Institute, University Medical Center, Rotterdam, The Netherlands.
Yunnan Key Laboratory of Chiral Functional Substance Research and Application, School of Chemistry & Environment, Yunnan Minzu University, Kunming, China.
PLoS One. 2024 Jun 7;19(6):e0304607. doi: 10.1371/journal.pone.0304607. eCollection 2024.
Hepatocellular carcinoma (HCC) is a highly aggressive liver cancer with significant morbidity and mortality rates. AXIN1 is one of the top-mutated genes in HCC, but the mechanism by which AXIN1 mutations contribute to HCC development remains unclear.
In this study, we utilized CRISPR/Cas9 genome editing to repair AXIN1-truncated mutations in five HCC cell lines.
For each cell line we successfully obtained 2-4 correctly repaired clones, which all show reduced β-catenin signaling accompanied with reduced cell viability and colony formation. Although exposure of repaired clones to Wnt3A-conditioned medium restored β-catenin signaling, it did not or only partially recover their growth characteristics, indicating the involvement of additional mechanisms. Through RNA-sequencing analysis, we explored the gene expression patterns associated with repaired AXIN1 clones. Except for some highly-responsive β-catenin target genes, no consistent alteration in gene/pathway expression was observed. This observation also applies to the Notch and YAP/TAZ-Hippo signaling pathways, which have been associated with AXIN1-mutant HCCs previously. The AXIN1-repaired clones also cannot confirm a recent observation that AXIN1 is directly linked to YAP/TAZ protein stability and signaling.
Our study provides insights into the effects of repairing AXIN1 mutations on β-catenin signaling, cell viability, and colony formation in HCC cell lines. However, further investigations are necessary to understand the complex mechanisms underlying HCC development associated with AXIN1 mutations.
肝细胞癌（HCC）是一种侵袭性很强的肝癌，发病率和死亡率都很高。AXIN1 是 HCC 中突变频率最高的基因之一，但 AXIN1 突变促进 HCC 发展的机制尚不清楚。
本研究利用 CRISPR/Cas9 基因组编辑技术修复了五株 HCC 细胞系中的 AXIN1 截断突变。
我们成功地从每株细胞系中获得了 2-4 个正确修复的克隆，这些克隆的 β-catenin 信号均减弱，细胞活力和集落形成能力降低。虽然修复的克隆暴露于 Wnt3A 条件培养基中可恢复 β-catenin 信号，但并未或仅部分恢复其生长特征，表明存在其他机制的参与。通过 RNA-seq 分析，我们探索了与修复的 AXIN1 克隆相关的基因表达模式。除了一些高响应的 β-catenin 靶基因外，未观察到基因/通路表达的一致改变。这一观察结果也适用于 Notch 和 YAP/TAZ-Hippo 信号通路，这些通路以前与 AXIN1 突变型 HCC 有关。AXIN1 修复的克隆也不能证实最近的观察结果，即 AXIN1 直接与 YAP/TAZ 蛋白稳定性和信号有关。
本研究深入了解了修复 AXIN1 突变对 HCC 细胞系中 β-catenin 信号、细胞活力和集落形成的影响。然而，需要进一步研究来理解与 AXIN1 突变相关的 HCC 发展的复杂机制。
