LINC00518 通过 miR-526b-3p/EIF5A2 轴影响皮肤恶性黑色素瘤细胞的增殖、侵袭和迁移。
LINC00518 affects the proliferation, invasion and migration of cutaneous malignant melanoma cells via miR-526b-3p/EIF5A2 axis.
机构信息
Plastic and Cosmetic Center, Affiliated Jinhua Hospital, Zhejiang University School of Medicine, Jinhua, Zhejiang 321000, P.R. China.
Department of Dermatology, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, P.R. China.
出版信息
Acta Biochim Pol. 2022 Feb 21;69(1):101-111. doi: 10.18388/abp.2020_5746.
OBJECTIVE
To investigate the mechanism of LINC00518 affecting the proliferation, invasion, and migration of cutaneous malignant melanoma (CMM) cells via miR-526b-3p/EIF5A2 axis.
METHODS
qRT-PCR was performed to measure the expression of LINC00518, miR-526b-3p, and EIF5A2 in CMM tissues from 40 patients. Si-LINC00518, pcDNA-LINC00518, miR-526b-3p mimic, miR-526b-3p inhibitor, si-EIF5A2, and their corresponding negative controls were transfected alone or co-transfected into CMM cells A375 and A2058. The expression of LINC00518, miR-526b-3p and EIF5A2 in A375 and A2058 cells was measured. Cell proliferation was tested by CCK-8 assay and EdU assay. Cell invasion and migration were detected by Transwell and scratch tests, respectively. The binding between LINC00518 and miR-526b-3p, and the binding between miR-526b-3p and EIF5A2 were verified by dual-luciferase reporter and RNA pull-down assays.
RESULTS
LINC00518 and EIF5A2 were up-regulated and miR-526b-3p was down-regulated in CMM tissues and cells. CMM patients with highly expressed LINC00518 showed decreased survival time than those with lowly expressed LINC00518. Transfection of si-LINC00518, miR-526b-5p mimic or si-EIF5A2 weakened the proliferative, migratory, and invasive abilities of melanoma cells, while transfection of miR-526b-5p inhibitor or pcDNA-LINC00518 enhanced the progression of melanoma cells. Moreover, the proliferative, migratory, and invasive potentials of melanoma cells were decreased after co-transfection of si-EIF5A2 and pcDNA-LINC00518 compared with cells transfected with pcDNA-LINC00518 alone. LINC00518 bound to miR-526b-3p and miR-526b-3p targeted EIF5A2. LINC00518 negatively regulated miR-526b-3p expression but positively regulated EIF5A2. Furthermore, EIF5A2 expression was negatively associated with miR-526b-3p expression.
CONCLUSION
LINC00518 encourages CMM through the miR-526b-3p/EIF5A2 axis in terms of cell proliferation, invasion, and migration.
目的
通过 miR-526b-3p/EIF5A2 轴研究 LINC00518 影响皮肤恶性黑色素瘤（CMM）细胞增殖、侵袭和迁移的机制。
方法
采用 qRT-PCR 法检测 40 例 CMM 组织中 LINC00518、miR-526b-3p 和 EIF5A2 的表达。单独转染或共转染 A375 和 A2058 细胞 Si-LINC00518、pcDNA-LINC00518、miR-526b-3p 模拟物、miR-526b-3p 抑制剂、si-EIF5A2 及其相应的阴性对照。检测 A375 和 A2058 细胞中 LINC00518、miR-526b-3p 和 EIF5A2 的表达。通过 CCK-8 检测和 EdU 检测测定细胞增殖。通过 Transwell 和划痕试验分别检测细胞侵袭和迁移。通过双荧光素酶报告和 RNA 下拉实验验证 LINC00518 与 miR-526b-3p 之间以及 miR-526b-3p 与 EIF5A2 之间的结合。
结果
LINC00518 和 EIF5A2 在 CMM 组织和细胞中上调，而 miR-526b-3p 下调。LINC00518 高表达的 CMM 患者的生存时间短于 LINC00518 低表达的患者。转染 si-LINC00518、miR-526b-5p 模拟物或 si-EIF5A2 减弱了黑素瘤细胞的增殖、迁移和侵袭能力，而转染 miR-526b-5p 抑制剂或 pcDNA-LINC00518 增强了黑素瘤细胞的进展。此外，与单独转染 pcDNA-LINC00518 的细胞相比，共转染 si-EIF5A2 和 pcDNA-LINC00518 后黑素瘤细胞的增殖、迁移和侵袭潜能降低。LINC00518 与 miR-526b-3p 结合，miR-526b-3p 靶向 EIF5A2。LINC00518 负调控 miR-526b-3p 的表达，但正调控 EIF5A2 的表达。此外，EIF5A2 的表达与 miR-526b-3p 的表达呈负相关。
结论
LINC00518 通过 miR-526b-3p/EIF5A2 轴促进 CMM 细胞的增殖、侵袭和迁移。
Zotero 插件