Lu Ruijuan, Zhang Xusheng, Li Xiaojuan, Wan Xuefeng
Department of Dermatovenereology, The First Affiliated Hospital of Northwest Minzu University (The Second People's Hospital of Gansu Province) Lanzhou, Gansu, China.
Department of General Surgery, The First Affiliated Hospital of Northwest Minzu University (The Second People's Hospital of Gansu Province) Lanzhou, Gansu, China.
Int J Clin Exp Pathol. 2020 Jul 1;13(7):1791-1801. eCollection 2020.
Malignant melanoma is a skin cancer with a high rate of metastasis. Numerous circular RNAs (circRNAs) have been shown to play vital roles in melanoma. This research aimed to investigate the role and molecular basis of circ_0016418 in melanoma progression.
The abundanced of circ_0016418, miR-605-5p and glutaminase (GLS) were measured using quantitative real-time polymerase chain reaction or western blot analysis. Cell proliferation was evaluated using Cell Counting Kit-8 (CCK-8) assay and colony formation assay. Cell migration and invasion were assessed by transwell assay. Cell cycle and apoptosis were monitored by flow cytometry. The levels of glutamine consumption and glutamate were examined using commercial kits. The interaction among circ_0016418, miR-605-5p and GLS was verified with the dual-luciferase reporter assay. A xenograft model was used to analyze tumor growth .
Circ_0016418 and GLS were up-regulated, while miR-605-5p was down-regulated in melanoma tissues and cells. Circ_0016418 silencing hindered cell proliferation, metastasis, and glutamine catabolism and promoted cell cycle arrest and apoptosis in A375 and A875 cells. Circ_0016418 modulated melanoma progression and glutamine catabolism through sponging miR-605-5p. Also, miR-605-5p inhibited melanoma progression and glutamine catabolism by targeting GLS. Moreover, circ_0016418 depletion blocked tumor growth .
Knockdown of circ_0016418 suppressed melanoma development and glutamine catabolism by modulating the miR-605-5p/GLS pathway.
恶性黑色素瘤是一种具有高转移率的皮肤癌。众多环状RNA（circRNA）已被证明在黑色素瘤中发挥重要作用。本研究旨在探讨circ_0016418在黑色素瘤进展中的作用及分子机制。
采用定量实时聚合酶链反应或蛋白质免疫印迹分析检测circ_0016418、miR-605-5p和谷氨酰胺酶（GLS）的表达水平。使用细胞计数试剂盒-8（CCK-8）法和集落形成试验评估细胞增殖。通过Transwell试验评估细胞迁移和侵袭。采用流式细胞术监测细胞周期和凋亡。使用商业试剂盒检测谷氨酰胺消耗和谷氨酸水平。通过双荧光素酶报告基因试验验证circ_0016418、miR-605-5p和GLS之间的相互作用。利用异种移植模型分析肿瘤生长情况。
circ_0016418和GLS在黑色素瘤组织和细胞中上调，而miR-605-5p下调。circ_0016418沉默抑制了A375和A875细胞的增殖、转移和谷氨酰胺分解代谢，并促进细胞周期停滞和凋亡。circ_0016418通过海绵吸附miR-605-5p调节黑色素瘤进展和谷氨酰胺分解代谢。此外，miR-605-5p通过靶向GLS抑制黑色素瘤进展和谷氨酰胺分解代谢。而且，circ_0016418缺失可阻止肿瘤生长。
敲低circ_0016418通过调节miR-605-5p/GLS通路抑制黑色素瘤发展和谷氨酰胺分解代谢。
