Dermatological Department, Nanxishan Hospital of Guangxi Zhuang Autonomous Region, Guilin City, China.
Department of Dermatology & STD, QingDao No.8 People's Hospital, Qingdao, China.
J Clin Lab Anal. 2022 Sep;36(9):e24630. doi: 10.1002/jcla.24630. Epub 2022 Aug 5.
LncRNAs are closely related to cutaneous melanoma (CM) tumorigenesis and metastasis, and it can affect the progression of CM by regulating cell proliferation, migration, invasion, apoptosis, and other cellular mechanisms. This study investigated the role of LINC00665 in CM.
Expressions of LINC00665, miR-339-3p, and tubulin beta chain (TUBB) in CM cells were analyzed by qRT-PCR and/or Western blot. The LINC00665/miR-339-3p/TUBB targeting network was predicted by bioinformatics tools, screened out by Venn diagrams and analyzed by Pearson's correlation coefficients, followed by validation via dual-luciferase reporter assay and/or pull-down assay. Transfection of siLINC00665 or miR-339-3p inhibitor/mimic was conducted with CM cells whose viability, proliferation, migration, invasion, cell cycle progression, and apoptosis were measured by CCK-8 assay, colony formation assay, wound healing assay, Transwell assay, and flow cytometry. The associations of TUBB with tumor biological characteristics and other proteins were analyzed by CanserSEA and String, respectively.
High-expressed LINC00665 was detected in CM cells. Silencing LINC00665 decreased CM cell viability; inhibited colony formation, cell cycle progression, migration and invasion; enhanced apoptosis; and upregulated miR-339-3p. LINC00665 targeted miR-339-3p which targeted TUBB. MiR-339-3p upregulation induced effects similar to the LINC00665-silencing-induced effects and could downregulate TUBB, which was associated with malignant behaviors and related to other five proteins. MiR-339-3p downregulation induced the opposite effects of what miR-339-3p upregulation induced, and the miR-339-3p downregulation-induced effects could be reversed by LINC00665 silencing.
Silencing LINC00665 inhibits in vitro CM progression and induces apoptosis via the miR-339-3p/TUBB axis.
长链非编码 RNA 与皮肤黑色素瘤(CM)的肿瘤发生和转移密切相关,通过调节细胞增殖、迁移、侵袭、凋亡等细胞机制影响 CM 的进展。本研究探讨了 LINC00665 在 CM 中的作用。
通过 qRT-PCR 和/或 Western blot 分析 CM 细胞中 LINC00665、miR-339-3p 和微管蛋白β链(TUBB)的表达。通过生物信息学工具预测 LINC00665/miR-339-3p/TUBB 靶向网络,通过 Venn 图筛选,并用 Pearson 相关系数进行分析,然后通过双荧光素酶报告基因检测和/或下拉实验进行验证。用 siLINC00665 或 miR-339-3p 抑制剂/模拟物转染 CM 细胞,通过 CCK-8 测定、集落形成测定、划痕愈合测定、Transwell 测定和流式细胞术测定细胞活力、增殖、迁移、侵袭、细胞周期进程和凋亡。分别通过 CanserSEA 和 String 分析 TUBB 与肿瘤生物学特征和其他蛋白质的相关性。
在 CM 细胞中检测到高表达的 LINC00665。沉默 LINC00665 降低了 CM 细胞的活力;抑制集落形成、细胞周期进程、迁移和侵袭;增强凋亡;上调 miR-339-3p。LINC00665 靶向 miR-339-3p,miR-339-3p 靶向 TUBB。miR-339-3p 的上调诱导的作用与 LINC00665 沉默诱导的作用相似,并下调 TUBB,与其他五种蛋白质相关。miR-339-3p 的下调诱导与 miR-339-3p 上调诱导的相反作用,并且 miR-339-3p 下调诱导的作用可以通过 LINC00665 沉默来逆转。
沉默 LINC00665 通过 miR-339-3p/TUBB 轴抑制体外 CM 进展并诱导细胞凋亡。