SBK2通过代谢重编程促进胃癌细胞增殖 和抗凋亡

SBK2 Promoting Proliferation and Anti-Apoptosis of Gastric Cancer Cells via Metabolic Reprogramming

摘要: 目的：探讨SH3结构域结合激酶2 (SBK2)基因在胃癌细胞凋亡、细胞周期调控、代谢重编程过程中的作用影响。方法：采用慢病毒载体介导的基因编辑技术，构建SBK2基因敲低(sh-SBK2)及过表达(OE-SBK2)的KATO III稳定转染细胞模型；应用流式细胞术系统分析SBK2表达水平对胃癌细胞凋亡率及细胞周期分布的调控作用；利用商品化检测试剂盒分析各组细胞ATP、葡萄糖含量及乳酸脱氢酶活性，评估SBK2的表达对胃癌细胞代谢重编程相关产物含量的变化。结果：(1) 流式细胞术检测结果显示，OE-SBK2显著抑制凋亡进程、shSBK2转染显著促进细胞凋亡(P < 0.001)。(2) 敲降SBK2导致G1期细胞比例增加、S期减少(P < 0.001)；而过表达SBK2组G1期细胞减少(P < 0.01)、S期明显增加(P < 0.001)。(3) 代谢相关指标检测结果显示，OE-SBK2组ATP、葡萄糖含量显著增加，乳酸脱氢酶活力也得到显著提升(P < 0.001)，而在敲减SBK2的胃癌细胞中，ATP、葡萄糖含量显著减少，乳酸活力也有下降趋势(P < 0.05)。结论：本项研究揭示了SBK2在胃癌中的促癌作用，并阐明了其通过调控代谢重编程来促进胃癌细胞增殖和抗凋亡的关键分子节点，提示SBK2可作为胃癌预测性生物标志物及靶向治疗候选分子。

Abstract: Objective: To explore the effects of the SH3 domain-binding kinase 2 (SBK2) gene on the apoptosis, cell cycle regulation, and metabolic reprogramming of gastric cancer cells. Methods: Using lentiviral vector-mediated gene editing technology, stable transfected KATO III cell models with SBK2 gene knockdown (sh-SBK2) and overexpression (OE-SBK2) were constructed. Flow cytometry (Annexin V/PI double staining method) was applied to systematically analyze the regulatory effects of SBK2 expression levels on the apoptosis rate and cell cycle distribution of gastric cancer cells. Commercial detection kits were used to analyze the contents of ATP, glucose, and the activity of lactate dehydrogenase in each group of cells, to evaluate the changes in the contents of products related to metabolic reprogramming of gastric cancer cells due to the expression of SBK2. Results: (1) The results of flow cytometry detection showed that OE-SBK2 significantly inhibited the apoptosis process, and sh-SBK2 transfection significantly promoted cell apoptosis (P < 0.001). (2) Knocking down SBK2 led to an increase in the proportion of cells in the G1 phase and a decrease in the S phase (P < 0.001); in the group with SBK2 overexpression, the number of cells in the G1 phase decreased (P < 0.01), and the number of cells in the S phase increased significantly (P < 0.001). (3) The results of detecting metabolic-related indicators showed that the contents of ATP and glucose in the OE-SBK2 group increased significantly, and the lactate activity was also significantly enhanced (P < 0.001). In contrast, in gastric cancer cells with SBK2 knockdown, the contents of ATP and glucose decreased significantly, and the lactate activity also showed a downward trend (P < 0.05). Conclusion: This study reveals the tumor-promoting role of SBK2 in gastric cancer and clarifies the key molecular nodes through which it promotes the proliferation and anti-apoptosis of gastric cancer cells by regulating metabolic reprogramming. It suggests that SBK2 can be used as a predictive biomarker for gastric cancer and a candidate molecule for targeted therapy.