DANCR通过AKT-GSK3β-Cyclin D1信号通路调控黑素瘤细胞活性机制的研究

李程彬 徐威龙 李婷 陈佳 舒茂国 贾晶

[摘要]目的：探索长链非编码（Long non-coding，Lnc）RNA DANCR（Anti-differentiation nc RNA）在黑素瘤组织中表达及其对黑素瘤细胞的作用及其作用机制。方法：检测黑素瘤和瘤旁组织中DANCR表达，及其与患者临床预后的关系。在敲低DANCR的黑素瘤细胞株A375和B16中通过MTT和克隆形成实验检测敲低DANCR对细胞活性影响，通过Western-blot实验检测p-AKT，p-GSK3β和Cyclin D1表达，并通过细胞流式检测细胞周期分布。通过过表达DANCR检测其是否通过AKT/GSK3β调控细胞活性。结果：DANCR在黑素瘤组织中表达上调，并与黑素瘤不良临床预后相关;敲低DANCR可抑制黑素瘤细胞活性，下调p-AKT、p-GSK3β和Cyclin D1表达;抑制AKT或GSK3β均可减弱DANCR过表达对细胞活性上调作用。结论：DANCR在黑素瘤中表达上调，激活AKT-GSK3β-Cyclin D1信号通路上调细胞活性，促进黑素瘤进展。

[关键词]LncRNA DANCR;黑素瘤;细胞活性;增殖;AKT;GSK3β;Cyclin D1

[中图分类号]R739.5    [文献标志码]A    [文章编号]1008-6455（2021）03-0086-04

DANCR was Upregulated in Melanoma Tissues and Promoted Cell Viability Via AKT-GSK3β-Cyclin D1 Signaling

LI Cheng-bin，XU Wei-long，LI Ting，CHEN Jia，SHU Mao-guo，JIA Jing

（Department of Plastic，Cosmetic and Maxilofacial Surgery，the First Affiliated Hospital of Xian Jiaotong University，Xian 710061，Shaanxi，China）

Abstract： Objective  To investigate the expression of long non-coding RNA DANCR in melanoma tissues， and the role and potential mechanism of DANCR in melanoma development. Methods  Detecting the expression of DANCR in melanoma and the adjacent tissues to analyze the relation of DANCR expression and clinical prognosis. Knocking DANCR down in melanoma cell lines， A375 and B16. Cell viability was detected using MTT and colony formation assays. Western blot assay detected the expression of p-AKT， p-GSK3β and Cyclin D1. Flow cytometry assay analyzed the distribution of cell cycle. DANCR-overexpressed cells were used to confirm that DANCR regulated cell viability via AKT/ GSK3β signaling. Results  DANCR expression was upregulated in melanoma tissues compared with the adjacent tissues， which was associated with poor clinical prognosis. Knocking DANCR down repressed cell viability and diminished expression of p-AKT， p-GSK3β and Cyclin D1. Inhibiting both p-AKT and p-GSK3β reversed DANCR overexpression induced enhancement of cell viability. Conclusion  DANCR was upregulated in melanoma cells and enhanced melanoma cell viability via modulating AKT-GSK3β-Cyclin D1 signaling.

Key words： LncRNA DANCR; melanoma; cell viability; proliferation; AKT; GSK3β; Cyclin D1

惡性黑素瘤（Melanoma）是恶性度最高的皮肤肿瘤。近年来，黑素瘤在我国发病率成倍增长[1]。一直以来，其易转移，易复发的特点，加之目前并无针对性的治疗方案，导致黑素瘤的愈后极差。小分子靶向药物和免疫治疗等新兴疗法延长了黑素瘤患者生存时间，但仍存在耐药现象[2]。因此，深入研究黑素瘤发生发展分子机制以寻找新的治疗靶点十分迫切。前期研究表明DANCR功能复杂，如对抗上皮组织细胞分化[3];抑制牙源干细胞和成骨细胞分化[4-5];在一定程度上与结肠癌不良预后相关[6]。此外，DANCR还可增加肝癌组织细胞干性，促进细胞生长和转移[7-8]。敲低DANCR可明显抑制乳腺癌细胞转移[9]。本研究以DANCR为研究对象，初步探索DANCR在黑素瘤中的作用及其可能作用机制。……