ROCK2调控骨肉瘤细胞的侵袭及迁移

目的 探讨骨肉瘤组织中ROCK2的表达对骨肉瘤细胞侵袭及迁移的影响及其作用机制。方法 采用实时荧光定量PCR、Western blot及免疫组织化学染色法检测骨肉瘤组织中ROCK2的表达；沉默骨肉瘤细胞中ROCK2的表达后，采用划痕愈合及Transwell实验分析骨肉瘤细胞的侵袭及迁移能力，并通过尾静脉转移实验检测降低ROCK2表达对体内转移的影响。检测骨肉瘤细胞中上皮间质转化（EMT）相关蛋白表达，并在下调ROCK2的骨肉瘤细胞中加入EMT诱导剂TGF-β，检测其对细胞侵袭及迁移的影响。结果 ROCK2在骨肉瘤组织中的表达显著高于其相应的癌旁组织（P<0.05），下调骨肉瘤细胞中ROCK2的表达后，其体内外侵袭及迁移能力明显被抑制（P<0.05）；EMT相关蛋白E-cadherin表达下降，而N-cadherin及Vimentin蛋白明显增加；诱导骨肉瘤细胞中EMT发生后能够减弱下调ROCK2对骨肉瘤细胞侵袭及迁移的抑制作用。结论 ROCK2在骨肉瘤组织中表达升高，通过诱导EMT的发生进而促进骨肉瘤细胞的侵袭及迁移。ROCK2可能是骨肉瘤靶向治疗的潜在生物标志物。

ROCK2 Regulates Invasion and Migration of Osteosarcoma Cells

Objective To analyze the expression of ROCK2 in osteosarcoma tissues and explore its effect on the invasion and migration of osteosarcoma cells and related mechanism.Methods Real-time fluorescent quantitative PCR,Western blot and immunohistochemical staining were used to detect the expression of ROCK2 in osteosarcoma tissues.The migration and invasion of osteosarcoma cells were analyzed by wound-healing and Transwell assays after the knockdown of ROCK2 expression.The effect of reducing ROCK2 expression on metastasis in vivo was tested by tail vein metastasis experiment.We detected the expression of EMT-related proteins in osteosarcoma cells,and added the EMT inducer TGF-β to osteosarcoma cells with down-regulated ROCK2 expression to detect the changes of cell invasion and migration.Results The expression of ROCK2 in osteosarcoma tissues was significantly higher than that in corresponding adjacent tissues (P<0.05).After down-regulating the expression of ROCK2 in osteosarcoma cells,the invasion and migration abilities in vivo and in vitro were significantly inhibited (P<0.05).The expression of EMT-related protein E-cadherin decreased,while N-cadherin and Vimentin proteins increased significantly.Inducing EMT could attenuate the inhibitory effect of down-regulating ROCK2 on the invasion and migration of osteosarcoma cells.Conclusion The expression of ROCK2 is increased in osteosarcoma tissues,which promotes the invasion and migration of osteosarcoma cells by inducing EMT.ROCK2 may be a potential biomarker for targeted therapy of osteosarcoma.