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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 elucidate the expression level of miR-139 in osteosarcoma cells and its effect on proliferation and apoptosis of osteosarcoma cells. Methods The expression of miR-139 was detected by qRT-PCR in immortalized osteoblasts hF OB 1.19 and osteosarcoma cell lines SOSP-9607 and MG-63. 5-Aza-2’-deoxycytidine (5Aza- CdR) and trichostatin A (TSA) were used to treat osteosarcoma cells. The expression of miR-139 was detected by qRT-PCR. The miR-139 mimic was transfected into osteosarcoma cells. The cell proliferation ability was detected by MTT assay. Changes were analyzed by flow cytometry, and changes in Cyclin D1 expression and Caspase-3 activation were detected by Western blot. Results Compared with the immortalized osteoblast cell line hF OB 1.19, the expression level of miR-139 in SOSP-9607 and MG-63 osteosarcoma cells was significantly decreased (P<0.01); 5-Aza-CdR to SOSP-9607 and MG The expression of miR-139 in osteosarcoma cells had no effect on T-63, but TSA could promote the expression of miR-139 (P<0.01). MiR-139 mimics could inhibit the proliferation of osteosarcoma cells and induce the apoptosis of osteosarcoma cells. And inhibit Cyclin D1 expression and promote the activation of Caspase3 (P <0.05 or P <0.01). Conclusion miR-139 is lowly expressed in osteosarcoma cells, and over-expression of miR-139 can inhibit osteosarcoma cell proliferation and induce apoptosis.