摘要
目的 探索miR-499靶向TGF-αmRNA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-αmRNA高表达(P<0.01)。miR-499靶向负调控TGF-αmRNA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-αmRNA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。
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 whichsuggests that miR-499 may inhibit the development of osteosarcoma.
作者
王磊
邱明宪
张慧荣
张金萍
赵静
康肖
张清泉
WANG Lei;QIU Ming-xian;ZHANG Hui-rong;ZHANG Jin-ping;ZHAO Jing;KANG Xiao;ZHANG Qing-quan(the Fourth People s Hospital of Hengshui,Hengshui 053000;Hengshui People s Hospital of Hebei Province,Hengshui 053000,China)
出处
《基础医学与临床》
2022年第8期1230-1236,共7页
Basic and Clinical Medicine
基金
衡水市市级科技计划(2020014068Z)。
