长链非编码 RNA LINC00847 通过调控 miR-133a-3p/TGFBR1 轴促进黑色素瘤进展。
LncRNA LINC00847 Accelerates Melanoma Progression by Regulating MiR-133a-3p/TGFBR1 Axis.
机构信息
Department of Aesthetic, Plastic and Burn Surgery, The Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai, 264000, China.
Department of Burn and Plastic Surgery, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, 250021, China.
出版信息
Comb Chem High Throughput Screen. 2024;27(8):1231-1241. doi: 10.2174/1386207326666230816113411.
AIMS
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.
MATERIALS AND 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.
CONCLUSION
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.
目的
越来越多的证据表明，长链非编码 RNA（lncRNA）在肿瘤发生中发挥调节作用。新鉴定的 lncRNA（LINC00847）的失调已涉及多种肿瘤。然而，lncRNA 在皮肤黑色素瘤中的表达和作用仍不清楚。因此，我们试图在本研究中研究 lncRNA 的表达和作用。
材料和方法
从 TCGA 数据库和临床招募的参与者的组织样本中定量测定 LINC00847 的表达水平。通过转染 si-LINC00847 或 si-NC 抑制细胞中的 LINC00847 表达。使用 RT-qPCR 测定 LINC00847 和 miR-133a-3p 的表达水平，并使用 Western blot 测定 TGFBR1 水平。通过生物信息学工具预测 LINC00847 与 miR-133a-3p 的靶向位点以及 miR-133a-3p 与 TGFBR1 的靶向位点，并通过双荧光素酶报告系统和 RNA 免疫沉淀证实。使用 CCK8 分别评估细胞增殖、侵袭和迁移能力，使用细胞集落形成、细胞划痕实验和 Transwell 实验评估细胞增殖、侵袭和迁移能力。
结果
在 TCGA 和临床队列中，皮肤黑色素瘤组织中 LINC00847 的表达异常上调，而非良性痣。此外，A375 和 SK-MEL-28 细胞中 LINC00847 的表达增加更为明显，而非正常表皮黑素细胞（HEMa-LP 细胞）。LINC00847 敲低可显著抑制皮肤黑色素瘤细胞的增殖、转移和伤口愈合率。此外，miR-133a-3p/TGFBR1 是 LINC00847 的下游靶标。LINC00847 在皮肤黑色素瘤细胞中负调控 miR-133a-3p 的表达。miR-133a-3p 抑制剂和 TGFBR1 载体转染均逆转了 LINC00847 沉默对皮肤黑色素瘤细胞的作用。
结论
LINC00847 在皮肤黑色素瘤中高表达，其过表达加速了皮肤黑色素瘤细胞的恶性肿瘤行为。miR-133a-3p/TGFBR1 轴参与了 LINC00847 在皮肤黑色素瘤中的作用。
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