Key Laboratory of Environmental Medicine Engineering, School of Public Health, Ministry of Education, Southeast University, 87 Dingjiaqiao Street, Nanjing, 210009, China.
J Transl Med. 2024 Apr 30;22(1):401. doi: 10.1186/s12967-024-05157-z.
The cancer microbiota was considered the main risk factor for cancer progression. We had proved that Fusobacterium periodonticum (F.p) was higher abundance in Esophageal cancer(EC)tissues. Bioinformation analysis found that BCT was a key virulence protein of F.p. However, little is known about the role and mechanism of BCT in EC. This study aimed to recognize the key virulence protein of F.p and explore the mechanism of BCT in promoting EC.
We constructed a eukaryotic expression vector and purified the recombinant protein BCT. CCK8 used to analyzed the activity of EC after treated by different concentration of BCT. UPLC-MS/MS and ELISA used to detect the metabonomics and metabolites. The ability of migration and invasion was completed by transwell assay. RT-QPCR, WB used to analyze the expression of relevant genes.
Our data showed that BCT was higher expression in EC tumor tissues (p < 0.05) and BCT in 20 µg/mL promoted the survival, invasion and migration of EC cells (EC109) (p < 0.05). Meanwhile, UPLC-MS/MS results suggested that BCT resulted in an augmentation of hypotaurine metabolism, arachidonic acid metabolism, glycolysis/gluconeogenesis, tryptophan metabolism, citrate cycle activity in EC109. The metabolic changes resulted in decreasing in glucose and pyruvate levels but increase in lactate dehydrogenase (LDH) activity and lactic acid (LA) as well as the expression of glucose transporter 1, Hexokinase 2, LDH which regulated the glycolysis were all changed (p < 0.05). The BCT treatment upregulated the expression of TLR4, Akt, HIF-1α (p < 0.05) which regulated the production of LA. Furthermore, LA stimulation promoted the expression of GPR81, Wnt, and β-catenin (p < 0.05), thereby inducing EMT and metastasis in EC109 cells.
Altogether, these findings identified that impact of BCT in regulation of glycolysis in EC109 and its involves the TLR4/Akt/HIF-1α pathway. Meanwhile, glycolysis increasing the release of LA and promote the EMT of EC109 by GPR81/Wnt/β-catenin signaling pathway. In summary, our findings underscore the potential of targeting BCT as an innovative strategy to mitigate the development of EC.
癌症微生物群被认为是癌症进展的主要风险因素。我们已经证明，牙周梭杆菌（F.p）在食管癌（EC）组织中的丰度更高。生物信息分析发现 BCT 是 F.p 的关键毒力蛋白。然而，关于 BCT 在 EC 中的作用和机制知之甚少。本研究旨在识别 F.p 的关键毒力蛋白，并探讨 BCT 促进 EC 的机制。
我们构建了真核表达载体并纯化了重组蛋白 BCT。CCK8 用于分析不同浓度 BCT 处理后 EC 的活性。UPLC-MS/MS 和 ELISA 用于检测代谢组学和代谢物。通过 Transwell 测定分析迁移和侵袭能力。RT-QPCR、WB 用于分析相关基因的表达。
我们的数据表明，BCT 在 EC 肿瘤组织中表达较高（p<0.05），20μg/mL 的 BCT 促进 EC 细胞（EC109）的存活、侵袭和迁移（p<0.05）。同时，UPLC-MS/MS 结果表明，BCT 导致 EC109 中的高丝氨酸代谢、花生四烯酸代谢、糖酵解/糖异生、色氨酸代谢、柠檬酸循环活性增强。代谢变化导致葡萄糖和丙酮酸水平降低，但乳酸脱氢酶（LDH）活性和乳酸（LA）升高，以及调节糖酵解的葡萄糖转运蛋白 1、己糖激酶 2、LDH 的表达均发生变化（p<0.05）。BCT 处理上调了 TLR4、Akt、HIF-1α 的表达（p<0.05），调节 LA 的产生。此外，LA 刺激促进了 GPR81、Wnt 和 β-连环蛋白的表达（p<0.05），从而诱导 EC109 细胞发生 EMT 和转移。
总之，这些发现表明 BCT 调节 EC109 中的糖酵解及其涉及 TLR4/Akt/HIF-1α 通路。同时，糖酵解增加 LA 的释放，并通过 GPR81/Wnt/β-连环蛋白信号通路促进 EC109 的 EMT。总之，我们的研究结果强调了靶向 BCT 作为减轻 EC 发展的创新策略的潜力。
