MiR-650通过靶向ING4促进骨肉瘤细胞增殖的研究

Study on miR-650 for promoting proliferation of osteosarcoma cells via targeting ING4

目的探讨骨肉瘤组织和细胞系中miR-650的表达及其在肿瘤形成中的作用机制。方法用实时荧光定量PCR的方法,检测并比较肿瘤组织及癌旁正常组织、骨肉瘤细胞系(MG63)及健康人成骨细胞(hFOB1.19)之间的miR-650表达情况;用MTT实验检测不同组细胞的增殖情况;用Western blot的方法检测调节生长抑制因子4(ING4)蛋白的表达情况。结果miR-650在骨肉瘤组织及细胞系中的表达显著高于癌旁正常组织及健康人成骨细胞;抑制miR-650的表达后,MG63细胞的增殖能力显著减弱。此外,miR-650表达减低后,ING4的表达显著升高,其中阴性对照组、乱序组、miR-650抑制剂组的ING4mRNA分别为1.00±0.16、1.08±0.14、5.35±0.32;蛋白表达分别为:0.62±0.06、0.59±0.12、2.45±0.20;miR-650抑制组与阴性对照组比较差异有统计学意义(P<0.05);而抑制ING4升高后,MG63细胞的增殖能力有一定的恢复。结论 miR-650可以通过降低ING4的表达促进MG63细胞的增殖,提示miR-650可能成为骨肉瘤治疗的新靶点。

Objective To investigate the expression of miR-650 in osteosarcoma tissue and cell lines and its action mechanism in the osteosarcoma formation.Methods The realtime fluorescence quantitative PCR was used to detect and compare the expressions of miR-650 between osteosarcoma tissue and normal tissue or between osteosarcoma cell lines(MG63)and normal human osteoblast cell lines(hFOB1.19);the MTT experiment was used to investigate the proliferation situation in different groups;Western blot was used to detect the ING4 protein expression.Results The expression of miR-650 in osteosarcoma tissue and MG63 cells was higher than that in normal tissue and human osteoblast cell;after inhibiting miR-650 expression,the proliferation ability of MG63 was significantly decreased.Moreover the expression of inhibitor of growth 4(ING4)was significantly increased when miR-650 was reduced,ING4 mRNA of negative control group,scramble group and miRNA group were 1.00±0.16,1.08±0.14 and 5.35±0.32 respectively;the ING4 protein expressions were 0.62±0.06,0.59±0.12 and 2.45±0.20 respectively;compared with negative control group,the difference was statistically significant(P<0.05);but after suppressing this elevation,the proliferation ability of MG63 cells had a certain recovery.Conclusion MiR-650 promotes MG63 proliferation via reducing ING4 expression,which suggests that miR-650 could be a new target of treating osteosarcoma.