基础医学与临床 ›› 2022, Vol. 42 ›› Issue (8): 1230-1236.doi: 10.16352/j.issn.1001-6325.2022.08.1230

• 研究论文 • 上一篇    下一篇

miR-499通过靶向TGF-α抑制人骨肉瘤细胞系Saos2迁移和促凋亡

王磊1, 邱明宪1, 张慧荣1, 张金萍1, 赵静1, 康肖1*, 张清泉2   

  1. 1.河北省衡水市第四人民医院, 河北 衡水 053000;
    2.河北省衡水市人民医院, 河北 衡水 053000
  • 收稿日期:2021-06-01 修回日期:2021-11-22 出版日期:2022-08-05 发布日期:2022-08-01
  • 通讯作者: *Kangxiao1980@163.com
  • 基金资助:
    衡水市市级科技计划 (2020014068Z)

miR-499 inhibits migration and promotes apoptosis of human osteosarcoma cell line Saos2 by targeting at TGF-α

WANG Lei1, QIU Ming-xian1, ZHANG Hui-rong1, ZHANG Jin-ping1, ZHAO Jing1, KANG Xiao1*, ZHANG Qing-quan2   

  1. 1. the Fourth People's Hospital of Hengshui, Hengshui 053000;
    2. Hengshui People's Hospital of Hebei Province,Hengshui 053000, China
  • Received:2021-06-01 Revised:2021-11-22 Online:2022-08-05 Published:2022-08-01
  • Contact: *Kangxiao1980@163.com

摘要: 目的 探索miR-499靶向TGF-α mRNA蛋白水平调控人骨肉瘤细胞系Saos2细胞迁移和凋亡参与骨肉瘤的发病机制。方法 RT-qPCR检测miR-499和TGF-α在骨肉瘤组织中的表达差异;采用双荧光素酶报告基因实验检测miR-499和TGF-α靶向关系;Transwell小室法和TUNEL凋亡实验检测Saos2细胞迁移和凋亡;构建骨肉瘤裸鼠移植瘤模型,检测miR-499/TGF-α轴在体内实验中的调控机制。结果 骨肉瘤组织,miR-499表达降低(P<0.01),而TGF-α mRNA高表达(P<0.01)。miR-499靶向负调控TGF-α mRNA蛋白表达(P<0.01)。转染miR-499模拟物能抑制Saos2细胞迁移和促进凋亡(P<0.01),而TGF-α则促进细胞迁移和抑制凋亡(P<0.01),同时miR-499能抑制TGF-α的促癌效果。结论 miR-499抑制TGF-α mRNA蛋白水平从而抑制骨肉瘤细胞的生物学功能,提示miR-499可能具有抑制骨肉瘤发展的作用。

关键词: 骨肉瘤, miR-499, TGF-α, Saos2

Abstract: Objective To explore the role of miR-499 targeting TGF-α mRNA protein in regulating the migration and apoptosis of human osteosarcoma cell line Saos2 and its pathogenesis. Methods The expression differences of miR-499 and TGF-α in osteosarcoma tissues were detected by RT-qPCR. Double luciferase reporter gene assay was used to detect the targeting relationship between miR-499 and TGF-α. Transwell cell assay and TUNEL apoptosis assay were used to detect the migration and apoptosis of Saos2 cells. A nude mouse xenograft model of osteosarcoma was constructed to detect the regulatory mechanism of miR-499/TGF-α axis in vivo. Results In osteosarcoma, miR-499 expression was decreased (P<0.01) but TGF-α mRNA expression was increased(P<0.01). miR-499 negatively regulated TGF-α mRNA protein expression (P<0.01). Transfection of miR-499 simulacrum inhibited the migration and promoted apoptosis of Saos2 cells (P<0.01), while TGF-α promoted the migration and inhibited apoptosis of Saos2 cells (P<0.01). miR-499 inhibited the carcinogenic effect of TGF-α. Conclusions miR-499 inhibits the biological function of osteosarcoma cell line Saos2 by inhibiting at TGF-α mRNA protein level which suggests that miR-499 may inhibit the development of osteosarcoma.

Key words: osteosarcoma, miR-499, TGF-α, Saos2

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