| miR-495-3p / WIF1 / Wnt 信号通路轴调节视网膜母细胞瘤细胞
增殖、 迁移和侵袭 |
| |
| 引用本文： | 洪博,崔蓓,田春雨,王丽君,王凤翔,曹利群.miR-495-3p / WIF1 / Wnt 信号通路轴调节视网膜母细胞瘤细胞
增殖、 迁移和侵袭[J].医学分子生物学杂志,2022,19(3):192-199. |
| |
| 作者姓名： | 洪博 崔蓓 田春雨 王丽君 王凤翔 曹利群 |
| |
| 作者单位： | 1解放军总医院第三医学中心眼科医学部 北京市, 100039
2陆军军医大学士官学校 石家庄市, 050081 |
| |
| 摘 要： | 目的 分析 miR-495-3p / Wnt 抑制因子 (Wnt inhibitor factor 1, WIF1) / Wnt 信号通路轴调节视网
膜母细胞瘤 (retinoblastoma, RB) 细胞增殖、 迁移和侵袭。 方法 生物信息学分析 WIF1 的差异表达及与
临床不良表型之间的关系, 并预测 miRNA 靶标; 双荧光酶素报告实验验证。 构建稳定过表达 WIF1 的 RB
细胞 (Vector 组、 WIF1 组)。 构建稳定过表达 miR-495-3p 的 RB 细胞 (miR-NC 组、 miR-495-3p 组和 miR495-3p + WIF1 组)。 Western 印迹检测 WIF1 蛋白及 Wnt 通路相关蛋白表达, 并进行体外功能试验。 裸鼠移
植瘤实验分析移植瘤体积重量。 免疫组化分析 WIF1 在 RB 组织中水平。 结果 WIF1 低表达, 与迁移、 侵袭
有关, 为 miR-495-3p 靶标, 双荧光酶素报告实验证实 WIF1 是 miR-495-3p 作用靶点。 WIF1 过表达抑制细胞增
殖、 迁移和侵袭, 影响细胞周期, 诱导凋亡。 体内实验显示 WIF1 在 RB 组织中过表达, 其过表达抑制瘤体生
长。 过表达 WIF1 下调 β-catenin、 c-Myc 蛋白水平 (P< 0. 05)。 过表达 miR-495-3p 下调 WIF1 蛋白水平, 上调
β-catenin 和 c-Myc 蛋白水平, 促进细胞增殖迁移、 迁移和侵袭, 抑制细胞凋亡 (P< 0. 05), 增加 WIF1 表达
可逆转 miR-495-3p 的作用 (P< 0. 05)。 结论 miR-495-3p 靶向 WIF1 通过 Wnt 信号通路抑制 RB 细胞增殖、
迁移和侵袭, 并诱导凋亡。
|
| 关 键 词： | 视网膜母细胞瘤 miR-495-3p 信号通路 |
miR-495-3p/WIF1/Wnt Signaling Pathway Regulates Proliferation,Migration and Invasion of Retinoblastoma Cells |
| |
| HONG Bo,CUI Bei,TIAN Chunyu,WANG Lijun,WANG Fengxiang,CAO Liqun.miR-495-3p/WIF1/Wnt Signaling Pathway Regulates Proliferation,Migration and Invasion of Retinoblastoma Cells[J].Journal of Medical Molecular Biology,2022,19(3):192-199. |
| |
| Authors: | HONG Bo CUI Bei TIAN Chunyu WANG Lijun WANG Fengxiang CAO Liqun |
| |
| Affiliation: | 1Department of Ophthalmology, The Third Medical Center, PLA General Hospital, Beijing,
100039, China
2Noncommissioned Officer School of Army Medical University, Shijiazhuang, 050081, China |
| |
| Abstract: | Objective To investigate the mechanism of miR-495-3p / Wnt inhibitor factor 1
(WIF1) / Wnt signaling pathway in regulating the proliferation, migration and invasion of retinoblastoma (RB) cells. Methods The relationship between differential expression of WIF1 in RB
patients and the clinically adverse phenotypes was analyzed by bioinformatics. The miRNAs which
bind with WIF1 were predicted and verified by dual-luciferase reporter assay. RB cells with stable
overexpression of WIF1 or miR-495-3p were constructed respectively. The expressions of WIF1 protein and Wnt pathway related proteins were detected by Western blotting. In vitro assays for cell proliferation, apoptosis, migration and invasion were conducted. The volume and weight of tumors were
measured in nude mice transplanted with xenograft tumors. The expression level of WIF1 in RB tissues was analyzed by immunohistochemistry. Results WIF1 was lowly expressed in RB tissues,
and the low expression of WIF1 was related to migration and invasion. WIF1 was the target of miR495-3p, which was verified by dual-luciferase reporter assay. Overexpression of WIF1 could inhibit
cell proliferation, migration and invasion, affect cell cycle, and induce apoptosis. In vivo assayshowed that the overexpression of WIF1 in RB tissues could inhibit tumor growth. The expression levels of β-catenin and c-Myc proteins were downregulated after WIF1 was overexpressed (P< 0. 05).
Overexpression of miR-495-3p could reduce the expression level of WIF1 protein, increase the expression levels of β-catenin and c-My, promote cell proliferation, migration and invasion, and inhibit apoptosis (P< 0. 05). The increased expression of WIF1 could reverse the effect of miR-495-
3p on proliferation, migration, invasion and apoptosis of RB cells (P< 0. 05). Conclusion miR495-3p could inhibit proliferation, migration and invasion of RB cells, and induce apoptosis by
WIF1 / Wnt signaling pathway. |
| |
| Keywords: | retinoblastoma miR-495-3p signaling pathway |
| 本文献已被 万方数据 等数据库收录！ |
| 点击此处可从《医学分子生物学杂志》浏览原始摘要信息 |
|
点击此处可从《医学分子生物学杂志》下载全文 |
|
