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Knockdown of circ_0084043 suppresses the development of human melanoma cells through miR-429/tribbles homolog 2 axis and Wnt/β-catenin pathway.

敲除circ_0084043 通过miR-4 2 9/tribbles同源物 2 轴和Wnt/β-catenin通路抑制人黑色素瘤细胞的发育。

  • 影响因子:3.40
  • DOI:10.1016/j.lfs.2020.117323
  • 作者列表:"Chen Z","Chen J","Wa Q","He M","Wang X","Zhou J","Cen Y
  • 发表时间:2020-02-15
Abstract

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/β-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 β-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/β-catenin signaling pathway.

摘要

目的: 环状rna (Circular RNAs,circRNAs) 已经成为多种肿瘤发生的新型调控因子,包括黑色素瘤。CircRNA_0084043 最近被证明在人黑色素瘤细胞中失调。然而,其在黑色素瘤中的作用和机制在很大程度上尚未揭示。 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同源物 2 (TRIB2) 的表达。分别用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐 (MTT) 法、流式细胞术和transwell法测定细胞增殖、凋亡、迁移和侵袭。Western blotting评价Wnt/β-catenin通路的激活。通过双荧光素酶报告基因检测和RNA免疫沉淀证实了circ_0084043 、miR-429 和TRIB2 之间的靶结合。在体内,生成小鼠异种移植模型来研究肿瘤生长。 关键发现: circ_0084043 和TRIB2 在人黑色素瘤组织和细胞系中表达上调。Circ_0084043 敲除和TRIB2 沉默均可降低A375 和SK-MEL-28 细胞的增殖、迁移和侵袭能力,但可促进细胞凋亡。此外,TRIB2 恢复部分消除了circ_0084043 敲除在体外黑色素瘤细胞中的肿瘤抑制作用。然后,我们验证了circ_0084043 通过海绵miR-429 积极和物理控制TRIB2 的表达。此外,当circ_0084043 被敲低时,A375 和SK-MEL-28 细胞 β-catenin、c-Myc和cyclinD1 的表达受到抑制,伴随着miR-429 的增加和trip2 的降低。值得注意的是,circ_0084043 下调在体内阻碍了A375 细胞的肿瘤生长。 意义: 敲除circ_0084043 可通过调节miR429/TRIB2 轴和灭活Wnt/β-catenin信号通路抑制黑色素瘤的恶性发展。

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影响因子:2.93
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影响因子:0.96
发表时间:2020-01-01
DOI:10.1097/DAD.0000000000001459
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皮肤肿瘤方向

皮肤肿瘤是发生在皮肤的细胞增生性疾病,是一种常见病。发生于皮内或皮下组织的新生物,种类很多,临床上分良性肿瘤和恶性肿瘤。恶性肿瘤可以不断增殖,引起转移,威胁生命,称为皮肤癌。

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