目的:越来越多的证据表明lncRNAs在肿瘤发生中起调节作用。新鉴定的lncRNA(LINC00847)的失调已经涉及几种肿瘤。然而,lncRNAs在皮肤黑色素瘤中的表达和作用尚不清楚.因此,我们试图研究lncRNAs在这项研究中的表达和作用。
方法:对来自TCGA数据库和临床招募的参与者的组织样品中LINC00847的表达水平进行定量。通过用si-LINC00847或si-NC转染抑制细胞中的LINC00847。应用RT-qPCR检测LINC00847和miR-133a-3p的表达,TGFBR1水平用Western印迹法测定。通过生物信息学工具预测了具有miR-133a-3p的LINC00847和具有TGFBR1的miR-133a-3p的靶向位点,并通过双荧光素酶报告系统和RNA免疫沉淀进行了证明。细胞增殖,入侵,和迁移能力使用CCK8,细胞集落形成,细胞伤口划痕,和transwell分析,分别。
结果:在TCGA和临床队列中,LINC00847在皮肤黑色素瘤组织中的表达比良性痣异常上调。此外,与正常表皮黑素细胞(HEMA-LP细胞)相比,在A375和SK-MEL-28细胞中LINC00847表达增加更显著。LINC00847敲低显著抑制皮肤黑素瘤细胞增殖,转移,和伤口愈合率。此外,miR-133a-3p/TGFBR1是LINC00847的下游靶标。LINC00847负调控皮肤黑素瘤细胞中miR-133a-3p的表达。miR-133a-3p抑制剂和TGFBR1载体转染均逆转了LINC00847沉默在皮肤黑色素瘤细胞中的作用。
结论:LINC00847在皮肤黑色素瘤中高表达,其过度表达加速了皮肤黑色素瘤细胞的恶性肿瘤行为。miR-133a-3p/TGFBR1轴参与LINC00847在皮肤黑色素瘤中的作用。
OBJECTIVE: Growing evidence has suggested that lncRNAs play a regulatory role in tumorigenesis. Dysregulation of a newly identified lncRNA (LINC00847) has been involved in several tumors. Nevertheless, the expression and roles of lncRNAs in skin melanoma remain unclear. Therefore, we attempted to investigate the expressions and roles of lncRNAs in this study.
METHODS: Expression levels of LINC00847 were quantified in tissue samples from the TCGA database and clinically recruited participants. LINC00847 was inhibited in cells by transfecting with si-LINC00847 or si-NC. Expressions of LINC00847 and miR-133a-3p were determined using RT-qPCR, and the
TGFBR1 level was determined using Western blotting. Targeting sites of LINC00847 with miR-133a-3p and miR-133a-3p with
TGFBR1 were predicted by bioinformatic tools and proved by dual-luciferase reporter system and RNA immunoprecipitation. Cell proliferation, invasion, and migration abilities were assessed using CCK8, cell colony formation, cell wound scratch, and transwell assay, respectively.
RESULTS: In both TCGA and clinical cohorts, the expression of LINC00847 was abnormally upregulated in skin melanoma tissues than that of benign nevus. Besides, LINC00847 expression increased more markedly in A375 and SK-MEL-28 cells than in normal epidermal melanocytes (HEMa-LP cells). LINC00847 knockdown remarkably restrained skin melanoma cell proliferation, metastasis, and wound healing rate. Furthermore, miR-133a-3p/
TGFBR1 was the downstream target for LINC00847. LINC00847 negatively regulated miR-133a-3p expression in skin melanoma cells. Both miR-133a-3p inhibitors and
TGFBR1 vector transfection reversed the effect of LINC00847 silence in skin melanoma cells.
CONCLUSIONS: LINC00847 was highly expressed in skin melanoma, and its overexpression accelerated the malignant tumor behavior of skin melanoma cells. The miR-133a-3p /
TGFBR1 axis was involved in the roles of LINC00847 in skin melanoma.