Knockdown of circ_0084043 suppresses the development of human melanoma cells through miR-429/tribbles homolog 2 axis and Wnt/β-catenin pathway
Knockdown of circ_0084043 suppresses the development of human melanoma cells through miR-429/tribbles homolog 2 axis and Wnt/β-catenin pathway
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DOI：
10.1016/j.lfs.2020.117323
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发表时间：
2020-02-15
期刊：
影响因子：
6.1
通讯作者：
Cen, Ying
中科院分区：
文献类型：
--
作者：
Chen, Zhibing;Chen, Junjie;Cen, Ying
目的：环状RNA（circRNA）已成为包括黑色素瘤在内的多种肿瘤发生的新型调控因子。CircRNA_0084043最近被证明在人黑素瘤细胞中是失调的。材料和方法：采用逆转录-定量PCR（RT-qPCR）和Western blotting检测circ_0084043、miRNA（miR）-429和tribbles homolog 2（TRIB 2）在黑色素瘤中的表达。采用MTT法、流式细胞仪和transwell法分别检测细胞增殖、凋亡、迁移和侵袭能力。Wnt/β-catenin通路的活化通过蛋白质印迹法评估。通过双荧光素酶报告基因测定和RNA免疫沉淀证实了circ_0084043、miR-429和TRIB 2之间的靶结合。在体内，产生小鼠异种移植模型以研究肿瘤生长。关键发现：circ_0084043和TRIB 2在人黑素瘤组织和细胞系中的表达上调。在A375和SK-MEL-28细胞中，circ_0084043基因敲低和TRIB 2基因沉默均能降低细胞增殖、迁移和侵袭能力，但促进细胞凋亡。此外，TRIB 2恢复部分消除了体外黑色素瘤细胞中circ_0084043敲低的肿瘤抑制作用。然后，我们验证了circ_0084043通过海绵状miR-429积极且物理地控制Trib 2表达。此外，circ_0084043基因敲除后，A375和SK-MEL-28细胞中β-catenin、c-Myc和cyclinD 1的表达受到抑制，同时伴随miR-429的增加和TRIB 2的减少。值得注意的是，circ_0084043的下调在体内抑制了A375细胞的肿瘤生长。意义：circ_0084043的敲低可能通过调节miR 429/TRIB 2轴和失活Wnt/β-catenin信号通路来抑制黑色素瘤的恶性发展。
Aims: Circular RNAs (circRNAs) have been emerged as novel regulators in multiple tumorigenesis, including melanoma. CircRNA_0084043 was recently demonstrated to be deregulated in human melanoma cells. Nevertheless, its role and mechanism are largely unrevealed in melanoma.Materials and methods: Expression of circ_0084043, miRNA (miR)-429 and tribbles homolog 2 (TRIB2) was detected using reverse transcription-quantitative PCR quantitative PCR (RT-qPCR) and western blotting. Cell proliferation, apoptosis, migration and invasion were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, flow cytometry and transwell assays, respectively. The activation of Wnt/beta-catenin pathway was evaluated by western blotting. The target binding among circ_0084043, miR-429 and TRIB2 was confirmed by dual-luciferase reporter assay and RNA immunoprecipitation. In vivo, mice xenograft model was generated to investigate tumor growth.Key findings: Expression of circ_0084043 and TRIB2 was upregulated in human melanoma tissues and cell lines. Both circ_0084043 knockdown and TRIB2 silencing could decrease cell proliferation, migration and invasion, but facilitate apoptosis in A375 and SK-MEL-28 cells. Furthermore, TRIB2 restoration partially abrogated the tumor-suppressive role of circ_0084043 knockdown in melanoma cells in vitro. Then, we verified that circ_0084043 positively and physically controlled TRIB2 expression through sponging miR-429. Besides, expression of beta-catenin, c-Myc and cyclinD1 was inhibited in A375 and SK-MEL-28 cells when circ_0084043 was knocked down, accompanied with increased miR-429 and decreased TRIB2. Notably, circ_0084043 downregulation impeded tumor growth of A375 cells in vivo.Significance: Knockdown of circ_0084043 suppressed the malignant development of melanoma presumably through modulating miR429/TRIB2 axis and inactivating Wnt/beta-catenin signaling pathway.