miR-139对骨肉瘤细胞增殖和凋亡的影响

Effects of miR-139 on osteosarcoma cell proliferation and apoptosis

目的阐明miR-139在骨肉瘤细胞中的表达水平及其对骨肉瘤细胞增殖和凋亡的影响。方法采用qRT-PCR检测miR-139在永生化成骨细胞系hF OB 1.19及骨肉瘤细胞系SOSP-9607、MG-63中的表达情况;采用5-氮杂-2'-脱氧胞苷(5Aza-CdR)和曲古抑菌素A(TSA)处理骨肉瘤细胞,通过qRT-PCR检测miR-139的表达变化;将miR-139模拟物转染入骨肉瘤细胞中,通过MTT实验检测细胞增殖能力的变化,通过流式细胞术分析细胞凋亡的变化,通过Western blot检测Cyclin D1表达和Caspase-3活化的变化。结果与永生化成骨细胞系hF OB 1.19相比,miR-139在SOSP-9607和MG-63骨肉瘤细胞中的表达水平显著降低(P<0.01);5-Aza-CdR对SOSP-9607和MG-63骨肉瘤细胞中miR-139的表达没有影响,而TSA能够促进miR-139的表达(P<0.01);miR-139模拟物能够抑制骨肉瘤细胞的增殖能力,诱导骨肉瘤细胞凋亡,并抑制Cyclin D1的表达、促进Caspase3的激活(P<0.05或P<0.01)。结论 miR-139在骨肉瘤细胞中低表达,而过表达miR-139能够抑制骨肉瘤细胞的增殖并诱导其凋亡。

Objective To clarify the expression level of miR-139 in human osteosarcoma cells and its effects on osteosarcoma cell proliferation and apoptosis. Methods The expression of miR-139 was detected by qRT-PCR in immortalized osteoblast cell line hF OB1. 19 and osteosarcoma cell lines SOSP-9607 and MG-63. Osteosarcoma cells were treated with 5-Aza- 2＇-deoxycytidine（ 5-Aza-CdR）and trichostatin A（ TSA）,and then miR-139 expression was detected by qRT-PCR. After miR-139 mimics was transfected into osteosarcoma cells,the cell proliferation ability,cell apoptosis rate,Cyclin D1 and active Caspase-3 expression were detected by MTT assay,flow cytometry and Western blot,respectively. Results Compared with hF OB 1.19 cells,miR-139 expression was significantly reduced in SOSP-9607 and MG-63 osteosarcoma cells（ P〈0. 01）. The expression of miR-139 was not affected by 5-Aza-CdR in SOSP-9607 and MG-63 osteosarcoma cells,while TSA promoted the expression of miR-139 in these cells（ P〈0. 01）. The miR-139 mimics was capable of inhibiting the proliferation,inducing cell apoptosis,suppressing Cyclin D1 expression,and promoting the activation of Caspase-3 in SOSP-9607 and MG-63 osteosarcoma cells（ P〈0. 05 or P〈0. 01）. Conclusion The expression of miR-139 is downregulated in human osteosarcoma cells and its overexpression can inhibit osteosarcoma cell proliferation and induce apoptosis.