PDF(Pubmed)
Abstract：
BACKGROUND: The Enoyl-CoA hydratase/isomerase family plays a crucial role in the metabolism of tumors, being crucial for maintaining the energy balance and biosynthetic needs of cancer cells. However, the enzymes within this family that are pivotal in gastric cancer (GC) remain unclear.
METHODS: We employed bioinformatics techniques to identify key Enoyl-CoA hydratase/isomerase in GC. The expression of ECHDC2 and its clinical significance were validated through tissue microarray analysis. The role of ECHDC2 in GC was further assessed using colony formation assays, CCK8 assay, EDU assay, Glucose and lactic acid assay, and subcutaneous tumor experiments in nude mice. The mechanism of action of ECHDC2 was validated through Western blotting, Co-immunoprecipitation, and immunofluorescence experiments.
RESULTS: Our analysis of multiple datasets indicates that low expression of ECHDC2 in GC is significantly associated with poor prognosis. Overexpression of ECHDC2 notably inhibits aerobic glycolysis and proliferation of GC cells both in vivo and in vitro. Further experiments revealed that overexpression of ECHDC2 suppresses the P38 MAPK pathway by inhibiting the protein level of MCCC2, thereby restraining glycolysis and proliferation in GC cells. Ultimately, it was discovered that ECHDC2 promotes the ubiquitination and subsequent degradation of MCCC2 protein by binding with NEDD4.
CONCLUSIONS: These findings underscore the pivotal role of the ECHDC2 in regulating aerobic glycolysis and proliferation in GC cells, suggesting ECHDC2 as a potential therapeutic target in GC.
METHODS: We employed bioinformatics techniques to identify key Enoyl-CoA hydratase/isomerase in GC. The expression of ECHDC2 and its clinical significance were validated through tissue microarray analysis. The role of ECHDC2 in GC was further assessed using colony formation assays, CCK8 assay, EDU assay, Glucose and lactic acid assay, and subcutaneous tumor experiments in nude mice. The mechanism of action of ECHDC2 was validated through Western blotting, Co-immunoprecipitation, and immunofluorescence experiments.
RESULTS: Our analysis of multiple datasets indicates that low expression of ECHDC2 in GC is significantly associated with poor prognosis. Overexpression of ECHDC2 notably inhibits aerobic glycolysis and proliferation of GC cells both in vivo and in vitro. Further experiments revealed that overexpression of ECHDC2 suppresses the P38 MAPK pathway by inhibiting the protein level of MCCC2, thereby restraining glycolysis and proliferation in GC cells. Ultimately, it was discovered that ECHDC2 promotes the ubiquitination and subsequent degradation of MCCC2 protein by binding with NEDD4.
CONCLUSIONS: These findings underscore the pivotal role of the ECHDC2 in regulating aerobic glycolysis and proliferation in GC cells, suggesting ECHDC2 as a potential therapeutic target in GC.
摘要：
背景：烯酰辅酶A水合酶/异构酶家族在肿瘤的代谢中起着至关重要的作用，对于维持癌细胞的能量平衡和生物合成需求至关重要。然而,该家族中在胃癌（GC）中至关重要的酶尚不清楚。
方法：我们使用生物信息学技术来鉴定GC中的关键Enoyl-CoA水合酶/异构酶。通过组织芯片分析验证ECHDC2的表达及其临床意义。使用集落形成测定法进一步评估ECHDC2在GC中的作用，CCK8测定，EDU检测,葡萄糖和乳酸测定，和裸鼠皮下肿瘤实验。ECHDC2的作用机制通过蛋白质印迹法得到验证,免疫共沉淀,和免疫荧光实验。
结果：我们对多个数据集的分析表明，GC中ECHDC2的低表达与不良预后显着相关。ECHDC2的过表达在体内和体外都显着抑制了GC细胞的有氧糖酵解和增殖。进一步的实验表明，ECHDC2的过表达通过抑制MCCC2的蛋白水平来抑制P38MAPK途径，从而抑制GC细胞的糖酵解和增殖。最终,发现ECHDC2通过与NEDD4结合促进MCCC2蛋白的泛素化和随后的降解。
结论：这些发现强调了ECHDC2在调节GC细胞有氧糖酵解和增殖中的关键作用，提示ECHDC2是GC的潜在治疗靶点。
方法：我们使用生物信息学技术来鉴定GC中的关键Enoyl-CoA水合酶/异构酶。通过组织芯片分析验证ECHDC2的表达及其临床意义。使用集落形成测定法进一步评估ECHDC2在GC中的作用，CCK8测定，EDU检测,葡萄糖和乳酸测定，和裸鼠皮下肿瘤实验。ECHDC2的作用机制通过蛋白质印迹法得到验证,免疫共沉淀,和免疫荧光实验。
结果：我们对多个数据集的分析表明，GC中ECHDC2的低表达与不良预后显着相关。ECHDC2的过表达在体内和体外都显着抑制了GC细胞的有氧糖酵解和增殖。进一步的实验表明，ECHDC2的过表达通过抑制MCCC2的蛋白水平来抑制P38MAPK途径，从而抑制GC细胞的糖酵解和增殖。最终,发现ECHDC2通过与NEDD4结合促进MCCC2蛋白的泛素化和随后的降解。
结论：这些发现强调了ECHDC2在调节GC细胞有氧糖酵解和增殖中的关键作用，提示ECHDC2是GC的潜在治疗靶点。
