摘要
目的:探讨下调肝受体类似物1(LRH1)对骨肉瘤U2OS细胞增殖、侵袭和迁移的影响及其机制。方法:采用RT-qPCR和Western blot分别检测人骨肉瘤细胞U2OS和成骨细胞hFOB1.19中LRH1 mRNA和蛋白表达差异;将体外培养的U2OS细胞分为对照组(Ctrl)、阴性对照(NC)-siRNA组和LRH1-siRNA组,RT-qPCR检测各组U2OS细胞中LRH1 mRNA表达,MTT、流式细胞术、Transwell小室和划痕实验分别检测各组U2OS细胞增殖活力、细胞周期分布情况、穿膜细胞数和划痕愈合率,Western blot检测各组U2OS细胞中LRH1和Wnt/β连环蛋白(β-catenin)信号通路相关蛋白β-catenin、细胞周期素D1(CyclinD1)、c-Myc、基质金属蛋白酶9(MMP-9)表达。结果:与hFOB1.19细胞相比,U2OS细胞中LRH1 mRNA和蛋白表达明显升高(P<0.05);与NC-siRNA组相比,LRH1-siRNA组U2OS细胞中LRH1 mRNA表达、细胞增殖活力、S期细胞占比、穿膜细胞数、划痕愈合率和LRH1、β-catenin、CyclinD1、c-Myc、MMP-9蛋白表达明显降低,而G0/G1期细胞占比明显升高(P<0.05);NC-siRNA组和Ctrl组上述指标差异无统计学意义(P>0.05)。结论:骨肉瘤U2OS细胞中LRH1表达上调,下调其表达可通过诱导细胞周期阻滞抑制骨肉瘤U2OS细胞增殖、减弱U2OS细胞侵袭和迁移,其作用机制可能与抑制Wnt/β-catenin信号通路活化有关。
Objective:To investigate effects of down-regulation of liver receptor homolog 1(LRH1)on proliferation,invasion and migration of osteosarcoma U2OS cells and its mechanism.Methods:mRNA and protein expression differences of LRH1 in human osteosarcoma cell line U2OS and osteoblast hFOB1.19 were detected by RT-qPCR and Western blot;U2OS cells were divided into control group(Ctrl),negative control(NC)-siRNA group and LRH1-siRNA group,expression of LRH1 mRNA in U2OS cells was detected by RT-qPCR,MTT assay,flow cytometry,Transwell chamber and scratch test were used to detect U2OS cell proliferation,cell cycle distribution,cell number and wound healing rate,expressions of LRH1 and Wnt/β-catenin signaling pathway related proteinsβ-catenin CyclinD1,c-Myc and matrix metalloproteinase-9(MMP-9) were detected by Western blot.Results:Compared with hFOB1.19 cells,expressions of LRH1 mRNA and protein in U2OS cells were significantly increased(P0.05).Conclusion:LRH1 is up-regulated in osteosarcoma U2OS cells.Down-regulation of LRH1 expression can inhibit proliferation of osteosarcoma U2OS cells and weaken invasion and migration of U2OS cells by inducing cell cycle arrest,whose mechanism may be related to inhibition of Wnt/β-catenin signaling pathway activation.
作者
曲野
刘立柱
徐瑞敏
吴开弟
代伟宏
QU Ye;LIU Lizhu;XU Ruimin;WU Kaidi;DAI Weihong(Department of Trauma and Orthopedics,the Second Affiliated Hospital of Hainan Medical University,Haikou 570311,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2022年第7期822-826,共5页
Chinese Journal of Immunology
基金
海南省卫生健康行业科研项目(19A200138)。
