| 王建华,宫晨,刘飞,等.下调OCT4基因对人骨肉瘤F5M2细胞系增殖能力的影响.骨科,2017,8(5): 394-398. |
| 下调OCT4基因对人骨肉瘤F5M2细胞系增殖能力的影响 |
| Effects of OCT4 down-regulation on proliferation of human osteosarcoma F5M2 cell line |
| 投稿时间：2017-06-30 |
| DOI：10.3969/j.issn.1674-8573.2017.05.012 |
| 中文关键词: 骨肉瘤 OCT4 细胞增殖 基因干扰 |
| 英文关键词: Osteosarcoma OCT4 Cell proliferation Gene interference |
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| 中文摘要: |
| 目的 探讨下调八聚体结合转录因子4（OCT4）基因对骨肉瘤细胞F5M2增殖能力的影响。方法 采用RNA干扰技术将OCT4基因特异性小干扰RNA（siOCT4）转染到处于对数生长期的F5M2骨肉瘤细胞中作为siOCT4组，并设立siNC组（转染siNC）及空白对照组。采用实时荧光定量PCR及蛋白免疫印迹法检测并比较siOCT4组和siNC组细胞中的OCT4 mRNA水平及蛋白表达水平；采用四甲基偶氮唑蓝（MTT）法及5-乙炔基-2′-脱氧尿苷（EdU）法检测并比较各组细胞的增殖能力；采用克隆形成试验检测并比较三组细胞的克隆形成能力。结果 siOCT4组中OCT4的mRNA表达水平及蛋白表达水平均较siNC组显著降低；MTT法的实验结果显示：与siNC组及空白对照组比较，siOCT4组细胞的生长曲线显著右移，增殖能力显著低于其他两组；EdU法检测结果显示：siNC组细胞的24 h平均增殖率为48.50%±4.54%，而siOCT4组细胞的24 h平均增殖率为32.30%±1.72%；与空白对照组及siNC组细胞比较，siOCT4组的克隆形成能力显著降低；以上差异均具有统计学意义（P均＜0.05）。结论 下调骨肉瘤细胞F5M2中OCT4的表达能抑制其细胞的增殖能力。 |
| 英文摘要: |
| Objective To investigate the effect of OCT4 down-regulation on the proliferation of F5M2 osteosarcoma cells. Methods The specific small interfering RNA of OCT4 (siOCT4) was transfected into osteosarcoma F5M2 cells in logarithmic growth phase by RNA interference as the RNA interference group, and the negative control group (transfected with siNC) and blank control group were established. The expression of OCT4 mRNA and protein in RNA interference group and negative control group was detected by q-PCR and Western blotting. MTT and EdU assays were used to detect the effect of OCT4 on the proliferation of F5M2 cells. The effect of the expression of OCT4 on cell clone formation was investigated by clonal formation assay. Results The mRNA and protein expression levels of OCT4 in RNA interference group were lower than those in negative transfection group. The results of MTT showed that as compared with the negative transfection group and the blank control group, the growth curve of the RNA interference group was shifted to the right, and the proliferation ability was significantly lower in the RNA interference group than the other two groups. The EdU test showed that the average proliferation rate in 24 h in the negative transfection group was 48.50%±4.54%, and that in the RNA interference group was 32.30%±1.72%. As compared with the blank control group and negative transfection group, the colony forming ability in the RNA interference group was significantly decreased. All the differences were statistically significant (P＜0.05 for all). Conclusion Down-regulation of the OCT4 expression in osteosarcoma cells can inhibit the proliferation of osteosarcoma cells. |
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