Sema6D对人骨肉瘤细胞增殖、迁移、侵袭及血管形成能力的影响及机制

Effect of Sema6D on Proliferation,Migration,Invasion and Angiogenesis-promoting Ability of Human Osteosarcoma Cells and Its Mechanism

目的探讨沉默信号素蛋白6D(Sema6D)对人骨肉瘤细胞增殖、迁移、侵袭及促进血管形成能力的影响。方法检测人骨肉瘤组织及细胞系中Sema6D的表达情况,脂质体法转染靶向siRNA后通过CCK-8、细胞划痕、Transwell、人脐静脉内皮细胞与肿瘤条件培养基共培养实验探究骨肉瘤细胞增殖、迁移、侵袭和体外促血管形成能力的改变。Western blot检测下游相关信号通路蛋白表达情况。结果Sema6D在人骨肉瘤组织及细胞系中相对高表达(P<0.05),沉默Sema6D后143B和MG63细胞的增殖、迁移及侵袭能力显著下降(P<0.05);人脐静脉内皮细胞体外血管形成能力减弱(P<0.01);PI3K/AKT/mTOR和ERK信号通路蛋白表达降低(P<0.05)。结论Sema6D在人骨肉瘤组织和细胞系中表达水平高,敲低骨肉瘤细胞Sema6D表达水平后可通过减弱PI3K/AKT/mTOR和ERK信号通路转导抑制细胞增殖、迁移、侵袭及促血管形成能力。

Objective To investigate the effect of Sema6D knockdown on the proliferation,migration,invasion and angiogenesis-promoting ability of human osteosarcoma cell lines.Methods The expression of Sema6D in clinical tissues and cell lines of human osteosarcoma was detected.After the targeted siRNA transfection,the changes of proliferation,migration and invasion were measured by CCK-8,wound healing and Transwell experiments.HUVECs were co-cultured with tumor conditioned medium to detect their tube formation ability.And the expression of signal pathway proteins was detected by Western blot.Results Sema6D was highly expressed in human osteosarcoma tissues and cell lines(P<0.05).After silencing Sema6D,the proliferation,migration and invasion of 143B and MG63 cells decreased significantly(P<0.05),the angiogenesis ability of HUVECs decreased in vitro(P<0.01),and the expression of PI3K/AKT/mTOR and ERK-related signal pathway proteins decreased(P<0.05).Conclusion Sema6D is overexpressed in human osteosarcoma tissues and cell lines.Knockdown of Sema6D expression level could inhibit the proliferation,migration,invasion and angiogenesis-promoting ability of human osteosarcoma cells via reducing PI3K/AKT/mTOR and ERK signal pathway.