转染和干扰Runx2基因对K7 M2细胞的影响

Effect of Transfection Runx2 and siRNA Runx2 on K7 M2

目的探讨干扰和转染Runx2基因后对K7M2骨肉瘤细胞体外生物学活性的影响。方法分别将阴性对照质粒、携带Runx2和siRNA Runx2的质粒转染到K7M2细胞中。应用Western blot检测转染和干扰Runx2基因对其蛋白表达的影响,并检测细胞体外的增殖能力(MTT)、侵袭性、迁移性和黏附性的变化,并应用流式细胞仪(FCM)检测相应细胞的凋亡率和细胞周期分布的变化。结果与空白对照组比,转染Runx2和siRunx2组的K7M2细胞中Runx2蛋白表达分别明显增强和减弱,而阴性对照质粒组没有明显改变。转染siRunx2组的细胞,MTT结果显示增殖能力明显下降,迁移性、侵袭性和增殖指数(PI)也明显下降,与空白对照组比差异有统计学意义(P<0.05),黏附性和凋亡率却没有明显变化。转染Runx2组K7M2的细胞增殖能力、迁移性、侵袭性和PI明显增高,与空白对照组比差异有统计学意义(P<0.05),黏附性和凋亡率没有明显变化。阴性对照组和空白对照组比上述指标没有明显改变。结论 Runx2可以促进K7M2细胞增殖,增强细胞的侵袭、迁移能力,其机制可能是和改变细胞周期的分布,使细胞更快的进入G2/M期有关。

Objective To investigate the effect of transfection Runx2 or siRNA Runx2 gene on the in vitro bio-characteristics of K7M2 osteosarcoma cell.Methods Scrambled plasmide,plasmids carrying Runx2 and siRNA Runx2 were transfected into K7M2 cells respectively.Runx2 protein expression after transfecting was detected with Western blot.The proliferation capacity,invasion,migration and adhesiveness were observed.Meanwhile,the apoptosis rate and cell cycle distribution of K7M2 cells in each group were detected using flow cytometry.Results Compared with control,Runx2 protein expression of K7M2 cells in the transfected or intervened Runx2 gene group were increased and reduced respectively.The proliferation capacity,migration,invasion and proliferative index in group transfected by siRNA Runx2 was decreased in comparison with control group,and the difference showed statistical deviation.However,the group transfected Runx2 showed opposite characteristics compared with control group.Neither adhesiveness nor apoptosis rate had obvious change in all three groups.There was no obvious change between the scrambled group and control group.Conclusion Runx2 can increase proliferation capacity,invasion,migration of K7M2.The potential mechanism is that it can change the cell cycle distribution,thus make cell transits to G2/M stage more quickly.