摘要
目的探究G蛋白偶联受体C型家族5A(GPRC5A)基因对骨肉瘤细胞系143B迁移和侵袭的影响及其机制。方法从数据集GSE32981筛选出转移瘤中差异表达的基因,结合TCGA数据库对差异表达基因进行生存分析;CCK8实验、划痕实验和Transwell小室法侵袭实验检测GPRC5A敲低或过表达对骨肉瘤细胞增殖、迁移和侵袭的影响;RT-qPCR和Western blot检测GPRC5A敲低或过表达对EMT相关基因mRNA和蛋白水平的影响。结果转移瘤中有299个基因表达水平发生改变,联合TCGA数据库进一步筛选出37个与骨肉瘤患者生存密切相关的基因;细胞实验结果表明,GPRC5A不影响骨肉瘤细胞增殖,GPRC5A过表达能显著促进骨肉瘤细胞迁移和侵袭(P<0.05);过表达GPRC5A能促进EMT相关蛋白N-Cad和Vim的表达,抑制E-Cad的表达(P<0.05)。结论GPRC5A能通过诱导EMT促进骨肉瘤细胞系143B迁移和侵袭,与骨肉瘤患者生存密切相关,可作为抗骨肉瘤转移药物潜在作用靶点。
Objective To investigate the effect and mechanism of GPRC5A(G protein-coupled receptor family C,member 5,group A)gene on the migration and invasion of osteosarcoma cell line 143B.Methods Differentially expressed genes in metastatic tumors were screened out from the GSE32981 data set,and the survival analysis of the differentially expressed genes was performed in combination with the TCGA database.CCK8 experiment,scratch experiment and Transwell invasion experiment were carried out to detect the effect of GPRC5A knockdown or overexpression on osteosarcoma cell proliferation,migration and invasion.Western blot and RT-qPCR detected the effect of GPRC5A knockdown or over-expression on protein and mRNA levels of EMT related genes.Results The expression of 299 genes was changed in metastatic osteosarcoma,and 37 genes closely related to the survival of osteosarcoma patients were further screened out combined with TCGA database.The results of cell experiments showed that GPRC5A had no effect on the proliferation of osteosarcoma cells,but GPRC5A over-expression could significantly promote osteosarcoma cells migration and invasion(P<0.05).GPRC5A overexpression promoted the expression of EMT-related proteins N-Cad and Vim,and inhibited the expression of E-Cad(P<0.05).Conclusions GPRC5A is closely related to the survival of osteosarcoma patients and may promote the migration and invasion of osteosarcoma cells by inducing EMT,so it is believed to be a potential target for anti-osteosarcoma metastasis drugs.
作者
宋奇
程安源
余铃
徐立
SONG Qi;CHENG An-yuan;YU Ling;XU Li(Department of Orthopedics,the First Hospital of Wuhan,Wuhan 430022;Department of Orthopedics,Renmin Hospital of Wuhan University,Wuhan 430060,China)
出处
《基础医学与临床》
CSCD
2020年第10期1335-1341,共7页
Basic and Clinical Medicine
基金
国家自然科学基金(81502575)。
