Department of Dermatology, Weihai Central Hospitai, Weihai 264400, China.
Department of Rheumatology, Weihai Central Hospitai, Weihai 264400, China.
Biomed Pharmacother. 2019 Dec;120:109438. doi: 10.1016/j.biopha.2019.109438. Epub 2019 Sep 18.
The aim was to study the mechanism of LncRNA FOXD3-AS1 in cutaneous melanoma.
FOXD3-AS1 levels in 47 pairs of melanoma samples were detected. We used qRT-PCR to detect FOXD3-AS1, miR-325 and MAP3K2 expression in different staging samples and cutaneous melanoma cell lines. We used Kaplan-Meier curve to analyze survival rate in patients with FOXD3-AS1 high and low expression. Sh-FOXD3-AS1, miR-325, miR-325 inhibitor and oeMAP3K2 were transfected. The proliferation of A375 and SK-MEL-1 was detected by CCK8 and EdU labeling assay and cell clone formation assay. Dual luciferase reporter assay and pull down assay was used to confirm the binding site of FOXD3-AS1, miR-325 and MAP3K2. Flow cytometry was applied to detect the effect of lncRNA on cell cycle. The migration and invasion ability were detected by transwell assay.
LncRNA FOXD3-AS1 highly expressed in cutaneous melanoma cells and tissues. Patients with highly expressed LncRNA FOXD3-AS1 were always with shorter overall survival time. When LncRNA FOXD3-AS1 was knockdown, proliferation, invasion and migration of cutaneous malignant melanoma, and tumor weight was inhibited, and cell cycle was arrested. LncRNA FOXD3-AS1 negatively regulated the expression of miR-325, and then improved the level of MAP3K2. MiR-325 was with similarly effects on above biological process, and MAP3K2 overexpression could rescue the influence of sh-FOXD3-AS1. Tumor volume and weight were measured to confirm the effect of sh-FOXD3-AS1 in vivo.
LncRNA FOXD3-AS1 could promote proliferation, invasion and migration of cutaneous malignant melanoma via regulating miR-325/MAP3K2 axis.
研究长链非编码 RNA FOXD3-AS1 在皮肤黑色素瘤中的作用机制。
检测 47 对黑色素瘤样本中 FOXD3-AS1 的水平。采用 qRT-PCR 检测不同分期样本和皮肤黑色素瘤细胞系中 FOXD3-AS1、miR-325 和 MAP3K2 的表达。采用 Kaplan-Meier 曲线分析 FOXD3-AS1 高表达和低表达患者的生存率。转染 Sh-FOXD3-AS1、miR-325、miR-325 抑制剂和 oeMAP3K2。通过 CCK8 和 EdU 标记检测和细胞克隆形成实验检测 A375 和 SK-MEL-1 的增殖。采用双荧光素酶报告基因实验和下拉实验验证 FOXD3-AS1、miR-325 和 MAP3K2 的结合位点。流式细胞术检测 lncRNA 对细胞周期的影响。Transwell 实验检测迁移和侵袭能力。
长链非编码 RNA FOXD3-AS1 在皮肤黑色素瘤细胞和组织中高表达。FOXD3-AS1 高表达的患者总生存时间总是较短。当 LncRNA FOXD3-AS1 被敲低时，皮肤恶性黑色素瘤的增殖、侵袭和迁移以及肿瘤重量受到抑制，细胞周期被阻滞。FOXD3-AS1 负调控 miR-325 的表达，进而提高 MAP3K2 的水平。miR-325 对上述生物学过程具有类似的作用，而 MAP3K2 的过表达可以挽救 sh-FOXD3-AS1 的影响。通过测量肿瘤体积和重量来确认 sh-FOXD3-AS1 在体内的作用。
LncRNA FOXD3-AS1 可通过调节 miR-325/MAP3K2 轴促进皮肤恶性黑色素瘤的增殖、侵袭和迁移。
