LPA受体和胞内信号传导相关分子拮抗骨肉瘤细胞黏附和迁移能力

摘要

目的:探讨溶血磷脂酸(LPA)受体和胞内信号传导相关分子拮抗骨肉瘤细胞的黏附和迁移能力.方法:培养骨肉瘤细胞Mg-63,传至第3代采用表面抗原鉴定,采用Transwell体外黏附和迁移实验检测骨肉瘤细胞Mg-63体外黏附和迁移能力的影响,并运用LPA受体及胞内信号传导趋化因子SDF-1α、MCP-1及其特异性阻断剂AMD3100、SB203580干预Mg-63,以明确骨肉瘤细胞Mg-63体外黏附和迁移的相关机制.结果:Transwell结果显示,第7天、14天,治疗组(LPA受体干预的Mg-63细胞)体外黏附和迁移能力与模型组(未经干预的细胞Mg-63)、对照组(正常成骨细胞)相比不断加强,差异有统计学意义(F=13.658,P<0.05;F=11.765,P<0.05).治疗后第21天,治疗组体外黏附和迁移能力与模型组相比不断减弱,差异无统计学意义(χ2=0.085,P>0.05).胞内信号传导趋化因子SDF-1α、MCP-1在不同时相均能促进骨肉瘤细胞Mg-63的黏附和迁移,差异有统计学意义(F=10.163,P<0.05),经LPA受体干预及特异性阻断剂AMD3100、SB203580处理不同时相后,黏附和迁移能力下降,差异有统计学意义(F=12.016,P<0.05).结论:LPA具有促细胞骨架变化进而调节细胞黏附和迁移能力的作用,拮抗骨肉瘤黏附和迁移过程中存在不同时相的Mg-63细胞体外黏附和迁移,机制可能与MCP-1/CCR2、SDF-1α/CXCR4信号轴关系密切.%Objective:To investigate the lysophosphatidic acid receptor( LPA) and intracellular signaling-related molecules in antagonizing the adhesion and migration ability of osteosarcoma cells.Methods:The osteosarcoma cell Mg-63 was cultured and passed to the third generation.The adhesion and migra-tion ability of Mg-63 was assayed by Transwell using in vitro technique.The expression of LPA receptor and intracellular signal transduction of chemokines ( SDF-1α,MCP-1) and its specific blockers ( AMD3100,SB203580) were used to investigate the mechanisms of adhesion and migration of osteosarcoma cells Mg-63 in vitro.Results:Transwell test showed that the adhesion and migration ability of Mg-63 cells in vitro was significantly higher than that in the model and control groups at day 7 and 14,and the difference was statistically significant(F=13.658,P<0.05;F=11.765,P<0.05).On the 21st day after treatment,the adhesion and migration ability of Mg-63 cells in vitro was weakened compared with the model group,yet the difference was not significant (χ2=0.085,P>0.05).Intracellular signal transduction chemotactic factor SDF-1α and MCP-1 promoted the adhesion and migration of osteosarcoma cell Mg-63 in different time phase,with statistical difference(F=10.163,P<0.05).After treatment with LPA receptor and specific blocker AMD3100 and SB203580 in diverse time phase,the adhesion and migration ability was decreased,and the difference was statistically significant (F=12.016,P<0.05). Conclusion:LPA has the effect of promoting cytoskeletal changes and regulating cell adhesion and migration ability,and antagonizing the adhesion and mi-gration of Mg-63 in vitro during the adhesion and migration of osteosarcoma.The mechanism may be related to SDF-1α / CXCR4,MCP-1 / CCR2 signal axis.