摘要
目的:探讨溶血磷脂酸受体-1(LPAR1)对骨肉瘤细胞增殖、迁移和侵袭的影响及其作用机制。方法:应用基因集富集分析(GSEA)方法分析LPAR1的表达与细胞恶性表型的关系。LPAR1质粒及其空载对照转染人骨肉瘤细胞MG63、U2OS。CCK-8法、流式细胞术、Transwell小室法检测细胞的增殖、凋亡、转移及侵袭能力。建立体内裸鼠移植瘤,观察LPAR1对瘤体生长的影响。Western blotting法检测mTOR/AKT信号通路激活的情况。结果:LPAR1在骨肉瘤中低表达;GSEA分析结果显示,负调控细胞增殖、周期、迁移和侵袭和肿瘤转移基因集富集以及细胞凋亡基因集富集在LPAR1高表达组。与空载体组相比,LPAR1组人骨肉瘤细胞MG63、U2OS的细胞活力、克隆形成率、迁移和侵袭能力及Ki67、CDK2、CCNB1、BCL2、pmTOR/mTOR、pAKT/AKT表达均明显下降,细胞凋亡率及BAX蛋白表达明显上升(P<0.05)。与空载体组相比,LPAR1组裸鼠的瘤体体积、质量明显下降,瘤体组织中的LPAR1蛋白表达上调(P<0.05)。结论:LPAR1在骨肉瘤中低表达,过表达LPAR1可抑制骨肉瘤细胞增殖、迁移和侵袭及裸鼠成瘤,其机制可能是通过抑制AKT信号通路来实现。
Objective:To explore the effects and mechanisms of lysophosphatidic acid receptor-1(LPAR1)on the proliferation,migration and invasion of osteosarcoma cells.Methods:Relationship between LPAR1 expression and malignant phenotypes was analyzed by gene set enrichment analysis(GSEA).Osteosarcoma cells MG63 and U2OS were transfected with LPAR1 plasmid and empty control.Proliferation,apoptosis,metastasis and invasion of cells were detected by CCK-8,flow cytometry and Transwell chamber assay,respectively.Xenograft tumors of nude mice were constructed to observe the effects of LPAR1 on tumors growth.Activation of mTOR/Akt signaling pathway was detected by western blotting.Results:LPAR1 was low-expressed in osteosarcoma.GSEA analysis showed that genes negatively regulated cell proliferation,cell cycle,migration and invasion,tumor metastasis and apoptosis were enriched in high-expressed LPAR1 group.Compared with empty vector group,the cell viability,clonal formation rate,migration and invasion of MG63 and U2OS cells,tumor volume and weight of nude mice,and the expressions of Ki67,CDK2,CCNB1,BCL2,pmTOR/mTOR and pAKT/AKT were significantly decreased in LPAR1 group,while the cell apoptosis rate and the protein expressions of LPAR1 and BAX were significantly increased(P<0.05).Conclusion:LPAR1 is low-expressed in osteosarcoma.LPAR1 overexpression can inhibit proliferation,migration and invasion of osteosarcoma cells as well as tumor formation in nude mice.The mechanism may be related to inhibiting AKT signaling pathway.
作者
吕俊杰
钟彩琴
张振宇
刘朋
姚鹏
Lyu Junjie;Zhong Caiqin;Zhang Zhenyu;Liu Peng;Yao Peng(Department of Orthopedics,Jinzhou Central Hospital,Jinzhou 121000,China;Department of Neurology,Jinzhou Central Hospital,Jinzhou 121000,China;Department of Orthopedics,The First Affiliated Hospital of Jinzhou Medical University,Jinzhou 121000,China)
出处
《广西医科大学学报》
CAS
2021年第12期2279-2285,共7页
Journal of Guangxi Medical University
基金
2018年度辽宁省自然科学基金立项项目(No.20180551208)。
关键词
骨肉瘤
溶血磷脂酸受体-1
AKT信号通路
增殖
迁移
侵袭
osteosarcoma
lysophosphatidic acid receptor-1
AKT signaling pathway
proliferation
migration
invasion
