miR-145 inhibits glutamine metabolism through c-myc/GLS1 pathways in ovarian cancer cells
miR-145 inhibits glutamine metabolism through c-myc/GLS1 pathways in ovarian cancer cells
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miR-145通过c-myc/GLS1途径抑制卵巢癌细胞中的谷氨酰胺代谢
DOI：
10.1002/cbin.11182
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发表时间：
2019-08-01
影响因子：
3.9
通讯作者：
Jiang, Yu
中科院分区：
文献类型：
--
作者：
Li, Jie;Li, Xu;Jiang, Yu
miR-145已被发现参与卵巢癌的肿瘤转移和糖代谢。然而，谷氨酰胺代谢在卵巢癌中的作用尚不清楚。在这项研究中，我们旨在阐明miR-145在卵巢癌细胞中调节谷氨酰胺代谢的分子机制。采用实时定量聚合酶链式反应（qRT-PCR）检测miR-145和谷氨酰胺酶1 （GLS1）的mRNA水平。western blot检测c-myc和GLS1蛋白水平。荧光素酶报告基因检测用于验证c-myc是miR-145的靶标。用检测试剂盒分析谷氨酰胺代谢。此外，我们进行了荧光素酶报告基因检测和染色质免疫沉淀检测，以验证c-myc转录激活GLS1并促进GLS1表达。qRT-PCR结果显示，miR-145与GLS1 mRNA水平在卵巢癌组织和细胞系中呈负相关。Kaplan-Meier生存分析和log-rank检验显示，miR-145高表达患者的总生存率显著提高。miR-145过表达抑制谷氨酰胺消耗、α -酮戊二酸生成和细胞ATP水平。此外，我们发现miR-145通过靶向c-myc抑制谷氨酰胺代谢。此外，c-myc可以通过转录激活促进GLS1的表达。总之，我们的研究结果揭示了miR-145在卵巢癌细胞中通过c-myc/GLS1途径抑制谷氨酰胺代谢，这可能会改善目前卵巢癌的诊断和治疗策略。
miR-145 has been found to be a participant in cancer metastasis and glucose metabolism in ovarian cancer. However, the role of glutamine metabolism in ovarian cancer remains unclear. In this study, we aim to elucidate the molecular mechanism underlying the regulation of glutamine metabolism by miR-145 in ovarian cancer cells. The messenger RNA (mRNA) levels of miR-145 and glutaminase 1 (GLS1) were examined by quantitative real-time polymerase chain reaction (qRT-PCR). The protein levels of c-myc and GLS1 were detected by western blot analysis. Luciferase reporter assays were used to validate c-myc was a target of miR-145. Glutamine metabolism was analyzed using assay kits. In addition, we performed luciferase reporter assays and chromatin immunoprecipitation assay to validate c-myc transcription activated GLS1 and promoted GLS1 expression. The qRT-PCR demonstrated that the mRNA level of miR-145 and GLS1 was negatively correlated in ovarian cancer tissues and cell lines. Kaplan-Meier survival analysis and the log-rank test showed that patients with high miR-145 expression had significantly increased the overall survival. The overexpression of miR-145 inhibited glutamine consumption, alpha-ketoglutarate production, and cellular ATP levels. Furthermore, we found miR-145 inhibited glutamine metabolism by targeting c-myc. Moreover, c-myc could promote GLS1 expression by transcription activated. Together, our results revealed that miR-145 inhibited glutamine metabolism through c-myc/GLS1 pathways in ovarian cancer cells, which may improve the current strategy of ovarian cancer diagnosis and therapy.