摘要
目的观察微小RNA（miRNA，miR）-422a表达水平对骨肉瘤细胞增殖抑制的影响。方法选择健康雄性SD大鼠40只，分别制备骨肉瘤MG63与U2OS细胞株荷瘤鼠模型，采用膜联蛋白V（Annexin V）/碘化丙锭（PI）双染色法转染miR-422a模拟物和抑制物调节miR-422a的表达，分析miR-422a高表达和低表达环境下骨肉瘤细胞的增殖、凋亡，并分析miR-422a表达对不同细胞周期骨肉瘤细胞的抑制作用。结果转染miR-422a模拟物后48 h，MG63和U2OS细胞的增殖出现明显抑制，在转染miR-422a抑制物48 h后，MG63和U2OS的细胞增殖被促进，差异均有统计学意义（P=0.068、0.035、0.016、0.004；0.072、0.028、0.019、0.003）；转染miR-422a模拟物后MG63细胞株对照组0、96 h的细胞凋亡率分别为（5.3±0.1）%、（5.2±0.1）%，模拟物组0、96 h的细胞凋亡率分别为（5.3±0.5）%、（5.4±0.4）%，U2OS细胞株对照组0、96 h的细胞凋亡率分别为（5.2±0.2）%、（5.3±0.2）%，模拟物组0、96 h的细胞凋亡率分别为（5.5±0.1）%、（5.2±0.1）%，转染miR-422a模拟物进行饥饿处理48 h后MG63细胞株对照组0、96 h的细胞凋亡率分别为（5.2±0.4）%、（28.6±0.7）%，模拟物组0、96 h的细胞凋亡率分别为（5.5±0.2）%、（48.5±2.4）%，U2OS细胞株对照组0、96 h的细胞凋亡率分别为（6.2±0.3）%、（18.7±3.1）%，模拟物组0、96 h的细胞凋亡率分别为（6.2±0.2）%、（39.8±3.2）%，与阴性对照组比较，转染miR-422a模拟物96 h后，骨肉瘤细胞的凋亡比例未出现显著改变，但在转染miR-422a模拟物进行诱导凋亡处理48 h后，与阴性对照组比较，骨肉瘤细胞凋亡数量明显增加，不同细胞生长周期细胞数量比较，转染miR-422a模拟物后G0/G1期细胞数明显增加，而S期和G2/M期细胞数明显降低。结论miR-422a高表达能够有效抑制骨肉瘤细胞的增殖，提高骨肉瘤细胞在恶劣环境中的凋亡比例。
Objective To observe the effect of microRNA （miRNA, miR）-422a expression on the proliferation of osteosarcoma cells.Methods 40 healthy male Sprague-Dawley （SD） rats were randomly divided into two groups： osteosarcoma MG63 and U2OS cell line bearing mice. Annexin V/propidium iodide （PI） double staining was used to transfect miR-422a primers and inhibitors to regulate miR-422a, the proliferation and apoptosis of osteosarcoma cells were analyzed under overexpression and miR-422a expression, and the inhibitory effect of miR-422a expression on different cell cycle osteosarcoma cells was examined.Results The proliferation of MG63 and U2OS cells was significantly inhibited at 48 h after transfection of miR-422a, and that of MG63 and U2OS was promoted at 48 h after transfection of miR-422a inhibitor with the difference being statistically significant （P=0.068, 0.035, 0.016, 0.004; 0.072, 0.028, 0.019, 0.003）. The apoptosis rate of MG63 cells in the control group at 0 and 96 h was （5.3±0.1）% and （5.2±0.1）% respectively, and that in miR-422a group was （5.3±0.5）% and （5.4±0.4）% respectively. The apoptosis rate of U20S in the control group at 0 and 96 h was （5.2±0.2）% and （5.3±0.2）% respectively, and that in miR-422a group was （5.5±0.1）% and （5.2±0.1）% respectively. After starvation for 48 h, the apoptosis rate of MG63 cells in control group at 0 and 96 h was （5.2±0.4）% and （28.6±2.7）% respectively, and that in miR-422a group was （5.5±0.2）% and （48.5±2.4）% respectively; the apoptosis rate of U20S in control group at 0 and 96 h was （6.2±0.3）% and （18.7±3.1）% respectively, and that in miR-422a group was （6.2±0.2）% and （39.8±3.2）% respectively. As compared with negative control group, the apoptosis rate of osteosarcoma cells had no significant changes in miR-422a group at 96 h. Howdever, the combination of miR-422a transfection and apoptosis induction for 48 h, the number of apoptotic osteosarcoma cells was significantly increased
出处
《中华实验外科杂志》
CSCD
北大核心
2017年第7期1185-1187,共3页
Chinese Journal of Experimental Surgery
