全反式维甲酸体外诱导分化对人骨肉瘤MG-63细胞侵袭转移的影响

Effect of ATRA on Invasion and Metastasis of Human Osteosarcoma MG-63 Cell during Differentiation Induction in vitro

目的观察全反式维甲酸(all trans retinoic acid,ATRA)诱导分化对人骨肉瘤MG-63细胞侵袭转移的抑制作用。方法体外用ATRA诱导MG-63细胞分化,通过电镜观察瘤细胞的超微结构;3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑嗅盐(MTT)检测细胞的增殖能力;软琼脂集落形成实验、侵袭和运动实验观察MG-63细胞生长和侵袭能力的变化;Western blot检测细胞增殖核抗原(proliferation cell nuclear antigen,PCNA)、细胞黏附分子44(cell differentiation44,CD44v6)蛋白的表达。结果经诱导剂处理后,细胞的生长明显受到抑制,细胞形态呈良性分化;肿瘤细胞的软琼脂集落形成率、穿膜运动能力均被显著抑制(P<0.01);与对照组相比随着作用时间的增加CD44v6和PCNA的表达均降低(P<0.01)。结论ATRA对人骨肉瘤MG-63细胞有诱导分化作用,并且从细胞的黏附、运动、降解等多环节抑制肿瘤细胞的侵袭转移能力。

Objective To observe the inhibitory effects of ATRA on invasion and metastasis of MG-63 cells during differentiation induction in vitro. Methods After in vitro differentiation induction by all trans retinoic acid （ATRA）, morphological and ultrastructural changes of MG-63 cells were observed under a transmission electron microscope. MTT assay was used to test the proliferation and protract cell growth curve. The cell growth and invasion ability was measured with the colony-formation rate in soft agar test, cell migration and invasion test. The expression of PCNA and CD44v6 protein was detected by Western blot. Results ATRA could significantly inhibil proliferation and induce morphodifferentiation of osteosarcoma cells, and the colony-formation rate in semi-solid agar and the invasion ability of MG-63 cells were decreased significantly （P〈0. 01 ）. The expression levels of CD44v6 and PCNA protein were decreased remarkably in osteosarcoma cells with the increase of treatment time. Conclusion ATRA can induce differentiation of human osteosareoma cells, and it also significantly inhibits the invasion and metastasis ability of MG-63 through cell adhesion, migration and invasion.