Background

Metastasis is a major cause of death in Colorectal cancer (CRC) patients, and the Epithelial–mesenchymal transition (EMT) has been known to be a crucial event in cancer metastasis. Downregulated expression of AT-rich interaction domain-containing protein 1A (ARID1A), a bona fide tumor suppressor gene, plays an important role in promoting EMT and CRC metastasis, but the underlying molecular mechanisms remain poorly understood. Here, we evaluated the impact of ARID1A knockdown and overexpression on the expression of EMT‑related genes, E-cadherin and β-catenin, in human CRC cells.

Methods and results

The expression levels of ARID1A, E-cadherin and β-catenin in CRC cell lines were detected via real-time quantitative PCR (qPCR) and western blot. ARID1A overexpression and shRNA-mediated knockdown were performed to indicate the effect of ARID1A expression on E-cadherin and β-catenin expression in CRC cell lines. The effect of ARID1A knockdown on the migration ability of HCT116 cells was assessed using wound-healing assay. We found that the mRNA and protein expression of adhesive protein E-cadherin was remarkably downregulated in response to shRNA-mediated ARID1A knockdown in HCT116 and HT29 cells. Conversely, overexpression of ARID1A in SW48 cells significantly increased E-cadherin expression. In addition, ARID1A silencing promoted the migration of HCT116 cells. ARID1A knockdown and overexpression did not alter the level of β-catenin expression.

Conclusions

Our study demonstrates that E-cadherin levels were closely correlated with ARID1A expression. Thus, ARID1A downregulation may promote CRC metastasis through decreasing EMT‑related protein E-cadherin and promoting epithelial cell movement. ARID1A could represent a promising candidate therapeutic target for CRC.

中文翻译：

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ARID1A 调节结直肠癌细胞中 E-cadherin 的表达：一个有希望的候选治疗靶点

背景

转移是结直肠癌（CRC）患者死亡的主要原因，已知上皮-间质转化（EMT）是癌症转移的关键事件。富含 AT 相互作用结构域的蛋白 1A (ARID1A) 的下调表达是一种真正的肿瘤抑制基因，在促进 EMT 和 CRC 转移中起重要作用，但其潜在的分子机制仍知之甚少。在这里，我们评估了ARID1A敲低和过表达对人 CRC 细胞中 EMT 相关基因E-钙粘蛋白和β-连环蛋白表达的影响。

方法和结果

通过实时定量PCR（qPCR）和蛋白质印迹检测CRC细胞系中ARID1A、E-cadherin和β-catenin的表达水平。进行 ARID1A过表达和 shRNA 介导的敲低以表明ARID1A表达对CRC 细胞系中E-钙粘蛋白和β-连环蛋白表达的影响。使用伤口愈合试验评估ARID1A敲低对 HCT116 细胞迁移能力的影响。我们发现粘附蛋白 E-cadherin 的 mRNA 和蛋白质表达在 HCT116 和 HT29 细胞中响应于 shRNA 介导的ARID1A敲低而显着下调。相反，ARID1A的过表达在 SW48 细胞中，E-cadherin 表达显着增加。此外，ARID1A沉默促进了 HCT116 细胞的迁移。ARID1A敲低和过表达不会改变β-连环蛋白的表达水平。

结论

我们的研究表明，E-cadherin 水平与ARID1A表达密切相关。因此，ARID1A下调可能通过减少 EMT 相关蛋白 E-钙粘蛋白和促进上皮细胞运动来促进 CRC 转移。ARID1A可能代表一个有希望的 CRC 候选治疗靶点。