MicroRNA-134在骨肉瘤中的表达及生物学作用研究

摘要

Objective To detect the expression of microRNA-134 in osteosarcoma and explore its biological function in osteosarcoma cell lines. Methods 40 cases of paired osteosarcoma tissues and adjacent matched normal osteosarcoma tissues were collected in our hospital and microRNA-134 expression was detected by using Quantitative RT-PCR analysis. The expres-sion levels of microRNA-134 was also analyzed in osteosarcoma cell lines( HOS and Saos-2)and a normal osteoblast cell (NHOst). The cells biological function was assessed by using MTT,wound-healing and Western blot assays,after transfected with microRNA-134 mimics into HOS and Saos-2 cells. Results The microRNA-134 expression in osteosarcoma tissues was significantly lower than paired normal tissues(P ﹤ 0. 01),and its expression was also down-regulated in HOS and Saos-2 cell lines than NHOst cell(P ﹤ 0. 05). Over-expression of microRNA-134 could markedly inhibit cell proliferation and migration, and promote BAX and CASPASE-3 expression. Conclusion MicroRNA-134 was significantly down-regulated in osteosarcoma tissues and cell lines. MicroRNA-134 can inhibit osteosarcoma cells proliferation and migration,and induce osteosarcoma cells apoptosis,which suggest that microRNA-134 function as a tumor suppressor gene involved in osteosarcoma development.%目的：检测微小 RNA -134（microRNA-134）在骨肉瘤的表达，并探究其在骨肉瘤细胞的生物学作用。方法收集40例骨肉瘤及癌旁正常骨组织标本，利用荧光定量 PCR（Quantitative RT-PCR）检测 microRNA-134的表达情况。进一步分析 microRNA-134在骨肉瘤细胞系（HOS 和 Saos-2）及正常成骨细胞系（NHOst）的表达，进而通过转染microRNA-134 mimics 至 HOS 和 Saos-2细胞，并应用四甲基偶氮唑盐微量酶反应比色法、细胞划痕以及凋亡试验探究其对骨肉瘤细胞的生物学作用。结果 MicroRNA-134在骨肉瘤组织标本中的表达量明显低于配对的癌旁组织（P ﹤0.01），同样 microRNA-134在 HOS 和 Saos-2细胞系中的表达亦低于 NHOst 细胞系（P ﹤0.05）。细胞生物学功能实验发现，过表达 microRNA-134可显著抑制 HOS 和 Saos-2细胞的增殖及迁移能力，并增加凋亡蛋白 Bcl-2相关 X 蛋白（BAX）及半胱氨酸天冬氨酸蛋白酶-3（CASPASE-3）表达。结论 MicroRNA-134在骨肉瘤组织及细胞系中明显低表达，并具有抑制骨肉瘤细胞增殖、迁移及增加凋亡的功能，高度提示其作为抑癌因子参与骨肉瘤的发生和发展过程。