[摘 要] 目的：探究β-半乳糖苷α-2-6唾液酸转移酶1（ST6GAL1）对结直肠癌（CRC）HCT116细胞糖酵解和迁移、侵袭的作用 及可能的分子机制。方法：通过检索GEPIA2数据库，分析ST6GAL1在CRC患者和健康人群中的表达差异；WB法检测 ST6GAL1在CRC细胞HCT116、SW480、Caco-2、HT29、LoVo和人正常结肠上皮细胞NCM460细胞中的表达差异；免疫组织化学 法分析ST6GAL1在CRC组织和对应癌旁组织中的表达差异。通过慢病毒转染细胞的方法构建稳定敲低或过表达ST6GAL1的 HCT116细胞，通过划痕愈合实验检测细胞迁移能力，Transwell实验检测细胞侵袭能力，WB法检测细胞糖酵解相关蛋白、Notch1 受体胞内段（Notch1 ICD）以及PI3K/AKT/mTOR通路磷酸化水平，细胞免疫荧光实验观察Notch1 ICD表达水平和进入细胞核情 况；加入Notch1受体激动剂Jagged1处理HCT116细胞，通过WB法检测糖酵解相关蛋白、Notch1 ICD表达水平以及PI3K/AKT/ mTOR通路磷酸化水平。结果：ST6GAL1在CRC组织和细胞中均表达上调（均P < 0.05）。与对照组和过表达组相比，敲低 ST6GAL1导致HCT116细胞内Notch1 ICD表达水平和PI3K/AKT/mTORC1磷酸化水平显著降低，细胞糖酵解相关蛋白表达水平 降低，细胞迁移和侵袭能力减弱（均P < 0.05）；过表达ST6GAL1增加了HCT116细胞内Notch1 ICD表达水平并促进其进入细胞 核，细胞糖酵解相关蛋白表达水平升高，细胞迁移和侵袭能力增强（均P < 0.05）。结论：ST6GAL1通过活化Notch1受体进而磷 酸化激活PI3K/AKT/mTORC1通路，并增强CRC细胞糖酵解水平和迁移、侵袭能力。

[Abstract] Objective: To explore the effect of β-galactoside α-2-6sialyltransferase1 (ST6GAL1) on glycolysis, migration and invasion of colorectal cancer (CRC) HCT116 cells and its possible molecular mechanisms. Methods: The difference in the expression of ST6GAL1 in CRC patients and healthy people was analyzed using the GEPIA2 database. WB was performed to detect the differences in the expressions of ST6GAL1 in CRC cell lines HCT116, SW480, Caco-2, HT29, LoVo and human normal colon epithelial cell line NCM460. The difference in the expressions of ST6GAL1 in CRC tissues and corresponding adjacent tissues was analyzed by immunohistochemistry. HCT116 cell lines with stably knocked down or overexpressed ST6GAL1 were constructed by lentivirus transfection. Cell migration ability was detected by scratch test. Cell invasion ability was detected by Transwell test. WB assay was performed to detect the expression levels of cell glycolysis related proteins and Notch1 intracellular domain (Notch1 ICD) as well as the phosphorylation level of PI3K/AKT/mTOR pathway. The expression level of Notch1 ICD and its entry into nucleus were observed by immunofluorescence assay. The Notch1 receptor agonist Jagged1 was added to HCT116 cells, and the expression levels of glycolysis-related proteins and Notch1 ICD and PI3K/AKT/mTOR pathway phosphorylation level were detected by WB. Results: The expression of ST6GAL1 was up-regulated in CRC tissues and cells (all P < 0.05). Compared with the control and overexpression groups, knockdown of ST6GAL1 resulted in significantly lower levels of Notch1 ICD expression and PI3K/AKT/mTORC1 phosphorylation in HCT116 cells, lower levels of cellular glycolysis-related protein expressions and weaker cell migration and invasion abilities (all P < 0.05). Overexpression of ST6GAL1 increased Notch1 ICD expression levels within HCT116 cells and promoted their entry into the nucleus. Cell glycolysis-related protein expression levels were elevated (all P < 0.05). Cell migration and invasion abilities were enhanced (all P < 0.05). Conclusion: ST6GAL1 activates the PI3K/AKT/mTORC1 pathway through activation of Notch1 receptor and phosphorylation, thus enhancing the glycolytic level and migration and invasion abilities of CRC cells.

新疆维吾尔自治区天池英才青年博士项目（No. 2025TCYCHYS）；新疆维吾尔自治区自然科学基金（No. 2021D01C185）