Qing Shunjie, Shen Zhiyong
Department of General Surgery, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2025 Mar 20;45(3):542-553. doi: 10.12122/j.issn.1673-4254.2025.03.12.
To explore the expression of hexokinase 2 (HK2) in colorectal cancer (CRC) and its possible mechanisms for regulating tumor cell behaviors and tumor immune microenvironment.
We analyzed HK2 expression in CRC and its impact on patient prognosis and tumor immune microenvironment using public databases. HK2 expression was also examined in 8 CRC and paired adjacent tissues using immunohistochemistry, Western blotting and RT-qPCR. In cultured CRC cell lines CT26 and HCT116 with low HK2 expression, the effects of lentivirus-mediated HK2 overexpression and JAK/STAT3 inhibitors on cell proliferation, migration, and invasion were assessed using CCK-8 assay, colony formation assay and Transwell assay and in a subcutaneous tumor-bearing mouse model; the changes were also observed in MC38 and CACO2 cells with high HK2 expressions following treatment with HK2 inhibitor 3-BP. Western blotting was performed to verify the relationship between HK2 and JAK/STAT signaling pathway protein expressions.
Informatics analyses suggested that HK2 expression was significantly higher in CRC tissues than in adjacent tissues (<0.001), and patients with high HK2 expressions had worse prognosis (=0.09). In the 8 clinical CRC tissues, HK2 expressions were significantly higher in the tumor tissues than in the adjacent tissues (<0.01). In CT26 and HCT116 cells, HK2 overexpression significantly enhanced cell proliferation, migration and invasion, while in HK2-overexpressing MC38 and CACO2 cells, inhibiting HK2 with 3-BP strongly suppressed these changes. HK2 overexpression promoted STAT3 phosphorylation, and JAK/STAT3 inhibitors effectively suppressed tumor cell proliferation, migration and invasion. TIMER and MCPcounter analyses indicated correlations between HK2 and immune cells, and TCGA and GEO analyses suggested significant positive correlations between HK2 and the immune checkpoints including PDCD1.
HK2 is upregulated in CRC to promote tumor cell proliferation, migration and invasion possibly by activating the JAK-STAT signaling pathway and modulating tumor immune microenvironment.
探讨己糖激酶2（HK2）在结直肠癌（CRC）中的表达及其调控肿瘤细胞行为和肿瘤免疫微环境的可能机制。
我们使用公共数据库分析了HK2在CRC中的表达及其对患者预后和肿瘤免疫微环境的影响。还采用免疫组织化学、蛋白质免疫印迹法和逆转录定量聚合酶链反应（RT-qPCR）检测了8对CRC组织及其配对的癌旁组织中HK2的表达。在HK2表达较低的CRC细胞系CT26和HCT116中，使用细胞计数试剂盒-8（CCK-8）检测法、集落形成试验和Transwell试验以及皮下荷瘤小鼠模型，评估慢病毒介导的HK2过表达和JAK/STAT3抑制剂对细胞增殖、迁移和侵袭的影响；在用HK2抑制剂3-溴丙酮酸（3-BP）处理后，观察HK2高表达的MC38和CACO2细胞中的变化。进行蛋白质免疫印迹法以验证HK2与JAK/STAT信号通路蛋白表达之间的关系。
信息学分析表明，CRC组织中HK2的表达显著高于癌旁组织（<0.001），HK2高表达的患者预后较差（=0.09）。在8例临床CRC组织中，肿瘤组织中HK2的表达显著高于癌旁组织（<0.01）。在CT26和HCT116细胞中，HK2过表达显著增强细胞增殖、迁移和侵袭，而在HK2过表达的MC38和CACO2细胞中，用3-BP抑制HK2可强烈抑制这些变化。HK2过表达促进信号转导和转录激活因子3（STAT3）磷酸化，JAK/STAT3抑制剂有效抑制肿瘤细胞增殖、迁移和侵袭。TIMER和MCPcounter分析表明HK2与免疫细胞之间存在相关性，癌症基因组图谱（TCGA）和基因表达综合数据库（GEO）分析表明HK2与包括程序性死亡受体1（PDCD1）在内的免疫检查点之间存在显著正相关。
HK2在CRC中上调，可能通过激活JAK-STAT信号通路和调节肿瘤免疫微环境来促进肿瘤细胞增殖、迁移和侵袭。
