Chen Chen, Liu Jing, Ma Yanli, Wang Yu, Cai Limin
Department of Dermatology, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang Province, 150001, China.
Heliyon. 2024 Aug 13;10(16):e36199. doi: 10.1016/j.heliyon.2024.e36199. eCollection 2024 Aug 30.
This study aimed to evaluate the effects of microRNA-650 (miR-650) on melanoma metastasis and reveal the regulatory relationship between miR-650 and the inhibitor of growth family member 4 (ING4).
miR-650 expression was determined in human melanoma WM115 and A-375 cells. WM115 cells were transfected with miR-650 mimic or mimic control. The invasion and migration abilities of transfected WM115 cells were analyzed using Transwell and wound healing assays, respectively. Then, miR-650-overexpression lentivirus vector was constructed and transfected into WM115 cells. After injection into the mice, the number of micro-metastatic foci in the lung tissues was counted. A regulatory relationship between miR-650 and ING4 was identified in WM115 and A-375 cells.
The miR-650 expression was upregulated in WM115 and A-375 cells. WM115 cells transfected with the miR-650 mimic exhibited higher invasive and migratory abilities than mock cells or cells transfected with negative control (NC). The number of micro-metastatic foci was significantly higher in mice injected with Lenti-miR-650 than that in those injected with mock or NC controls. Transfection with miR-650 mimic observably inhibited the expression of ING4 in WM115 and A-375 cells, whereas transfection with miR-650 inhibitor had the opposite effect. Dual-luciferase reporter gene assay showed that the miR-650 mimic inhibited the luciferase activity of ING4.
miR-650 promotes melanoma metastasis by downregulating ING4 expression.
本研究旨在评估微小RNA - 650（miR - 650）对黑色素瘤转移的影响，并揭示miR - 650与生长抑制家族成员4（ING4）之间的调控关系。
测定人黑色素瘤WM115和A - 375细胞中miR - 650的表达。用miR - 650模拟物或模拟对照转染WM115细胞。分别采用Transwell和伤口愈合试验分析转染后WM115细胞的侵袭和迁移能力。然后，构建miR - 650过表达慢病毒载体并转染至WM115细胞。注射到小鼠体内后，计数肺组织中的微转移灶数量。在WM115和A - 375细胞中鉴定miR - 650与ING4之间的调控关系。
WM115和A - 375细胞中miR - 650表达上调。用miR - 650模拟物转染的WM115细胞比未处理细胞或用阴性对照（NC）转染的细胞表现出更高的侵袭和迁移能力。注射Lenti - miR - 650的小鼠肺组织中的微转移灶数量明显高于注射未处理或NC对照的小鼠。用miR - 650模拟物转染可显著抑制WM115和A - 375细胞中ING4的表达，而用miR - 650抑制剂转染则产生相反的效果。双荧光素酶报告基因检测表明，miR - 650模拟物抑制了ING4的荧光素酶活性。
miR - 650通过下调ING4表达促进黑色素瘤转移。
