Objective: To investigate the expression of DARS2 and its clinical significance in colorectal cancer. Methods: In this study, bioinformatics tools, especially gene expression profile interactive analysis 2 (GEPIA2), were used to conduct an in-depth analysis of DARS2 expression in colorectal cancer tissues. Immunohistochemical staining was carried out in 108 colorectal cancer specimens and 30 normal colorectal tissues obtained from the First Affiliated Hospital of Nanchang University, Nanchang, China. Colorectal cancer cell lines (HCT116 and SW480) were transfected with small interfering RNA (siRNA) and DARS2 overexpression plasmid to examine the effects of DARS2 knockdown and overexpression on cell function. To assess the effects on cell function, CCK8 and transwell migration assays were used to assess proliferation and cell motility, respectively. Additionally, protein immunoblotting was employed to scrutinize the expression of proteins associated with the epithelial-mesenchymal transition of colorectal cancer cells. Results: DARS2 exhibited a pronounced upregulation in expression within colorectal cancer tissues compared to their normal epithelial counterparts. Furthermore, DARS2 expression was higher in colorectal cancer of stage Ⅲ-Ⅳ than those of stage Ⅰ-Ⅱ, exhibiting a significant correlation with N staging, M staging, and pathological staging (P<0.05). Kaplan-Meier analyses showed a decreased overall survival rate in colorectal cancer with DARS2 expression compared to those without DARS2 expression (P<0.05). In the siRNA transfection group, there was a significant reduction in cell proliferation and migration (P<0.01 and P<0.05, respectively). Conversely, the transfection of DARS2 overexpression plasmids substantially increased both cell proliferation and migration (P<0.05). Additionally, immunoblotting revealed that DARS2 knockdown led to an upregulation of E-cadherin expression and a downregulation of N-cadherin and vimentin expression. In contrast, DARS2 overexpression resulted in increased N-cadherin and vimentin expression, coupled with reduction in E-cadherin expression. Conclusions: There is a strong association between DARS2 expression and colorectal cancer progression. Silencing DARS2 inhibits cell proliferation and migration, exerting a discernible influence on the epithelial-mesenchymal transition process.
目的： 探讨结直肠癌中DARS2的表达情况及其临床意义。 方法： 采用生物信息学工具基因表达谱交互分析版本2（gene expression profiling interactive analysis 2，GEPIA2）分析DARS2在结直肠癌组织中的表达情况。采用免疫组织化学方法检测南昌大学第一附属医院的108例结直肠癌组织和30例正常结直肠上皮组织中DARS2的表达情况。以结直肠癌细胞HCT116和SW480为研究对象，通过将小干扰RNA（siRNA）或DARS2过表达质粒转染到细胞中，以检测敲低和过表达DARS2对细胞功能的影响。使用CCK8检测试剂盒检测DARS2对结直肠癌细胞增殖能力的影响，Transwell小室迁移实验检测DARS2对结直肠癌细胞迁移能力的影响，使用蛋白质免疫印迹检测结直肠癌细胞中上皮-间质转化相关蛋白的表达。 结果： 与正常结直肠上皮组织相比，DARS2在结直肠癌组织中的表达显著上调。DARS2在Ⅲ~Ⅳ期结直肠癌组织表达较Ⅰ~Ⅱ期高，临床基线数据表明DARS2表达与N分期、M分期和病理分期相关（P<0.05）。Kaplan-Meier生存曲线分析显示，DARS2高表达的结直肠癌患者具有较低的总体生存率（P<0.05）。在siRNA转染组检测到细胞增殖能力显著下降（P<0.01），细胞迁移能力也显著下降（P<0.05），而在转染DARS2过表达质粒后细胞增殖和迁移能力均显著增强（P<0.05）。同时，蛋白质免疫印迹结果显示在敲低DARS2后，上皮相关分子E-cadherin的表达上调，而间质相关分子N-cadherin和波形蛋白的表达下调，而过表达DARS2后N-cadherin和波形蛋白的表达上调、E-cadherin的表达下调。 结论： DARS2高表达与结直肠癌进展相关，敲低DARS2可以抑制细胞增殖与迁移，并影响上皮-间质转化过程。.