Abstract：
OBJECTIVE: Glutamine metabolism is crucial in cell proliferation, aging, and apoptosis across various cancer types. Existing research indicates that Sirtuin 4 (SIRT4), primarily located in mitochondria, modulates this process. This study aimed to clarify the regulatory relationship between SIRT4 and glutamine metabolism in cervical cancer.
METHODS: SIRT4 mRNA levels and their clinical correlation to cervical cancer were analyzed using the UALCAN database. Immunohistochemistry (IHC) was performed to assess SIRT4 protein expression in tissue samples from cervical cancer patients. Transient transfection was employed to create Hela and Siha cell lines with overexpressed SIRT4, mitogen-activated extracellular signal-regulated kinase (MEK), and glutaminase 1 (GLS1). The impact on cellular functions was studied using MTT, soft agar, transwell, and western blotting assays. Glutamate and ATP levels were also measured to evaluate metabolic changes.
RESULTS: Low levels of SIRT4 mRNA in cervical cancer tissues correlated with tumor metastasis and poor survival rates. Overexpression of SIRT4 led to suppressed cell proliferation, colony growth, and motility, along with significant down-regulation of GLS expression, a key contributor to glutamine metabolism. Additionally, SIRT4 overexpression resulted in the inactivation of the MEK/ERK/c-myc signaling pathway, while overexpression of MEK reversed these effects. Notably, the inhibitory effects of SIRT4 on cell proliferation, colony formation, migration, and invasion in Hela and Siha cells were significantly attenuated following GLS1 overexpression.
CONCLUSIONS: SIRT4 acts as an anti-cancer agent in cervical cancer by inhibiting glutamine metabolism through the MEK/ERK/c-myc signaling pathway, providing a novel sight for cervical cancer therapy.
METHODS: SIRT4 mRNA levels and their clinical correlation to cervical cancer were analyzed using the UALCAN database. Immunohistochemistry (IHC) was performed to assess SIRT4 protein expression in tissue samples from cervical cancer patients. Transient transfection was employed to create Hela and Siha cell lines with overexpressed SIRT4, mitogen-activated extracellular signal-regulated kinase (MEK), and glutaminase 1 (GLS1). The impact on cellular functions was studied using MTT, soft agar, transwell, and western blotting assays. Glutamate and ATP levels were also measured to evaluate metabolic changes.
RESULTS: Low levels of SIRT4 mRNA in cervical cancer tissues correlated with tumor metastasis and poor survival rates. Overexpression of SIRT4 led to suppressed cell proliferation, colony growth, and motility, along with significant down-regulation of GLS expression, a key contributor to glutamine metabolism. Additionally, SIRT4 overexpression resulted in the inactivation of the MEK/ERK/c-myc signaling pathway, while overexpression of MEK reversed these effects. Notably, the inhibitory effects of SIRT4 on cell proliferation, colony formation, migration, and invasion in Hela and Siha cells were significantly attenuated following GLS1 overexpression.
CONCLUSIONS: SIRT4 acts as an anti-cancer agent in cervical cancer by inhibiting glutamine metabolism through the MEK/ERK/c-myc signaling pathway, providing a novel sight for cervical cancer therapy.
摘要：
目的：谷氨酰胺代谢在细胞增殖中至关重要，老化，以及各种癌症类型的细胞凋亡。现有研究表明，Sirtuin4(SIRT4)，主要位于线粒体，调节这个过程。本研究旨在阐明SIRT4与谷氨酰胺代谢在宫颈癌中的调控关系。
方法：使用UALCAN数据库分析SIRT4mRNA水平及其与宫颈癌的临床相关性。进行免疫组织化学(IHC)以评估来自宫颈癌患者的组织样品中的SIRT4蛋白表达。瞬时转染用于创建具有过表达的SIRT4，丝裂原激活的细胞外信号调节激酶（MEK）的Hela和Siha细胞系，和谷氨酰胺酶1(GLS1)。使用MTT研究了对细胞功能的影响，软琼脂,transwell,和蛋白质印迹分析。还测量了谷氨酸和ATP水平以评估代谢变化。
结果：宫颈癌组织中SIRT4mRNA水平低与肿瘤转移和低生存率相关。SIRT4的过表达导致细胞增殖受到抑制,菌落生长，和运动性，随着GLS表达的显著下调，谷氨酰胺代谢的关键因素。此外,SIRT4过表达导致MEK/ERK/c-myc信号通路失活,而MEK的过度表达逆转了这些效应。值得注意的是,SIRT4对细胞增殖的抑制作用，菌落形成,迁移,GLS1过表达后，Hela和Siha细胞的侵袭能力显着减弱。
结论：SIRT4通过MEK/ERK/c-myc信号通路抑制谷氨酰胺代谢，在宫颈癌中发挥抗癌作用，为宫颈癌的治疗提供了新的视野。
方法：使用UALCAN数据库分析SIRT4mRNA水平及其与宫颈癌的临床相关性。进行免疫组织化学(IHC)以评估来自宫颈癌患者的组织样品中的SIRT4蛋白表达。瞬时转染用于创建具有过表达的SIRT4，丝裂原激活的细胞外信号调节激酶（MEK）的Hela和Siha细胞系，和谷氨酰胺酶1(GLS1)。使用MTT研究了对细胞功能的影响，软琼脂,transwell,和蛋白质印迹分析。还测量了谷氨酸和ATP水平以评估代谢变化。
结果：宫颈癌组织中SIRT4mRNA水平低与肿瘤转移和低生存率相关。SIRT4的过表达导致细胞增殖受到抑制,菌落生长，和运动性，随着GLS表达的显著下调，谷氨酰胺代谢的关键因素。此外,SIRT4过表达导致MEK/ERK/c-myc信号通路失活,而MEK的过度表达逆转了这些效应。值得注意的是,SIRT4对细胞增殖的抑制作用，菌落形成,迁移,GLS1过表达后，Hela和Siha细胞的侵袭能力显着减弱。
结论：SIRT4通过MEK/ERK/c-myc信号通路抑制谷氨酰胺代谢，在宫颈癌中发挥抗癌作用，为宫颈癌的治疗提供了新的视野。
