Twist调控骨肉瘤细胞的增殖、迁移和侵袭

摘要

目的 探讨增强或降低Twist表达对不同恶性程度的骨肉瘤细胞体外增殖、迁移、侵袭及体内成瘤的作用.方法 Real-time PCR及Western blot检测Twist在143B、MG63及TE85三种不同骨肉瘤中的基础表达.采用Ad-Twist及Ad-siTwist腺病毒感染细胞,细胞生长曲线检测细胞增殖能力,划痕实验检测细胞的平行迁移能力,Transwell实验检测细胞的垂直迁移及侵袭能力.Ad-Twist及Ad-siTwist感染Luc标记的143B细胞,制备胫骨近端骨肉瘤模型,活体成像动态观察细胞在体内的生长.结果在骨肉瘤细胞中Twist的基础表达显著高于C3H10细胞(P<0.05),其中在143B细胞表达最高,MG63细胞次之,TE85细胞中最低,Twist的表达水平与骨肉瘤细胞的恶性程度呈正相关.Ad-Twist及Ad-siTwist感染细胞分别能显著提高或降低骨肉瘤细胞中Twist的mRNA及蛋白水平的表达(P<0.05).生长曲线结果显示Twist过表达能显著促进骨肉瘤细胞的增殖,而抑制Twist的表达后,骨肉瘤细胞的增殖能力降低(P<0.05).Transwell实验发现Twist过表达后,3种骨肉瘤细胞迁移或侵袭至小室的细胞数明显增多,而抑制Twist的表达可降低骨肉瘤细胞的迁移侵袭能力(P<0.05).143B细胞具有良好的体内成瘤能力,Twist过表达促进143B细胞在裸鼠体内生长,活体成像动态观察荧光信号显著高于对照组,抑制Twist表达可显著抑制细胞在裸鼠体内生长,信号明显弱于对照组.结论 Twist可增强骨肉瘤细胞的增殖、迁移、侵袭及体内成瘤能力.%Objective To investigate the role of Twist in regulating the proliferation, migration, and invasion of osteosarcoma cells with different levels of malignancy. Methods The baseline expressions of Twist in 3 different osteosarcoma cell lines (143B,MG63 and TE85)were detected using real-time PCR and Western blotting.The cells were infected with the recombinant adenoviruses Ad-Twist or Ad-siTwist for Twist overexpression or knockdown,respectively,and the cell growth curves were drawn to assess the cell proliferation.The migration abilities and invasiveness of the cells were evaluated using wound healing assay and Transwell assay.Luc-labeled 143B cells infected with Ad-Twist or Ad-siTwist were intrathecally injected to establish nude mouse models bearing osteosarcoma xenografts, in which the tumor formation was monitored using living body imaging technique.Results The baseline expressions of Twist in the 3 osteosarcoma cells were significantly higher than that in C3H10 cells(P<0.05).Twist expression was the highest in 143B cells followed by MG63 cells,and was the lowest in TE85 cells, indicating its positive correlation with the level of malignancy of the osteosarcoma cells. Ad-Twist or Ad-siTwist infection efficiently enhanced or lowered Twist expressions at both mRNA and protein levels in osteosarcoma cells (P<0.05). Twist overexpression resulted in enhanced proliferation, migration and invasion abilities of osteosarcoma cells, and Twist knockdown obviously inhibited the cell proliferation, migration and invasion. In nude mice, 143B cells with Twist overexpression showed accelerated tumor formation compared with the control cells, while Twist knockdown significantly inhibited the tumor formation ability of the cells.Conclusion Twist overexpression can promote the proliferation,migration, invasion and tumorigenicity of osteosarcoma cells.