Department of Pediatrics, Yichang Yiling Hospital, China.
Department of Ultrasound Imaging, Yichang Yiling Hospital, China.
Mol Cell Probes. 2020 Jun;51:101540. doi: 10.1016/j.mcp.2020.101540. Epub 2020 Feb 18.
LncRNA MALAT1 is involved in regulation of angiogenesis, however, its expression and mechanism in infantile hemangioma (IH) are less reported. The study aimed to investigate MALAT1 in IH and to reveal the potential mechanism of MALAT1 acting on IH.
Isolated form IH tissue, human CD31 hemangioma endothelial cells (HemECs) were cultured and sorted by magnetic-activated cell sorting (MACS). Quantitative real-time (qRT)-PCR was performed to detect the expressions of MALAT1, miR-206 and VEGFA. The correlations among MALAT1, miR-206 and VEGFA were confirmed by bioinformatics analysis and dual-luciferase reporter assay. The effects of MALAT1, miR-206 and VEGFA on cell proliferation were detected by cell counting kit-8 (CCK-8) and cell colony formation assay. Flow cytometry, wound scratch, Transwell and Tube formation assay were performed to determine cell apoptosis, migration, invasion and vasoformation, respectively. Apoptosis-related proteins were determined by Western blot.
The results showed that MALAT1 and VEGFA were high-expressed and miR-206 was low-expressed in IH tissues. SiMALAT1 negatively affected the cell proliferation, migration, invasion and vasoformation of HemECs and promoted apoptosis of HemECs. Moreover, Bcl-2 expression was significantly inhibited and the expressions of Bax and c cleaved-3 were greatly promoted. MALAT1 directly targeted and inhibited the expression of miR-206, and VEGFA was predicted to be the target gene for miR-206. SiMALAT1 suppressed the cell proliferation, migration, invasion and vasoformation of HemECs through modulating miR-206/VEGFA axis.
Knock-down of MALAT1 inhibits the growth of HemECs through regulating miR-206/VEGFA axis, indicating that MALAT1 is a potential therapeutic mechanism for the treatment of IH.
长链非编码 RNA MALAT1 参与血管生成的调节，但其在婴儿血管瘤（IH）中的表达和机制报道较少。本研究旨在探讨 MALAT1 在 IH 中的作用，并揭示 MALAT1 作用于 IH 的潜在机制。
分离自 IH 组织的人 CD31 血管内皮细胞（HemECs）通过磁激活细胞分选（MACS）进行培养和分选。采用定量实时（qRT）-PCR 检测 MALAT1、miR-206 和 VEGFA 的表达。通过生物信息学分析和双荧光素酶报告基因检测证实 MALAT1、miR-206 和 VEGFA 之间的相关性。通过细胞计数试剂盒-8（CCK-8）和细胞集落形成实验检测 MALAT1、miR-206 和 VEGFA 对细胞增殖的影响。流式细胞术、划痕实验、Transwell 和管形成实验分别用于测定细胞凋亡、迁移、侵袭和血管生成。通过 Western blot 测定凋亡相关蛋白。
结果显示，IH 组织中 MALAT1 和 VEGFA 高表达，miR-206 低表达。SiMALAT1 可负向影响 HemECs 的细胞增殖、迁移、侵袭和血管生成，并促进 HemECs 凋亡。此外，Bcl-2 表达明显受到抑制，Bax 和 c cleaved-3 的表达显著上调。MALAT1 直接靶向并抑制 miR-206 的表达，而 VEGFA 被预测为 miR-206 的靶基因。SiMALAT1 通过调节 miR-206/VEGFA 轴抑制 HemECs 的细胞增殖、迁移、侵袭和血管生成。
敲低 MALAT1 通过调节 miR-206/VEGFA 轴抑制 HemECs 的生长，表明 MALAT1 是治疗 IH 的潜在治疗机制。
