CLEC4M对脑星形胶质母细胞瘤的增殖和迁移的影响

Study on CLEC4M in promoting the proliferation and migration of brain astroblastoma

目的:探讨C型凝集素蛋白家族4成员M(CLEC4M)对脑星形胶质母细胞瘤增殖和迁移的影响及其作用机制。方法:慢病毒转染法构建过表达和干扰CLEC4M的脑星形胶质母细胞瘤细胞系;并用实时定量PCR法检测CLEC4M的转录水平。CCK-8细胞毒性实验和细胞划痕实验分别检测CLEC4M对细胞增殖和迁移的影响,流式细胞术分析CLEC4M对细胞周期的影响。转录组测序技术鉴定癌症信号通路上被CLEC4M调控的基因。结果:过表达和干扰CLEC4M脑星形母细胞瘤细胞株被成功构建,且过表达和干扰的效果显著(P<0.01)。CLEC4M促进脑星形胶质母细胞瘤的增殖和迁移,且CLEC4M主要影响G0/G1和S期来调控细胞周期进程(P<0.05~P<0.01)。此外,转录组测序结果发现在脑星形胶质母细胞瘤中过表达CLEC4M导致5857个基因在转录水平上有明显变化,其中2897个基因显著上调,2960个基因显著下调。KEGG富集通路分析显示这些差异表达的基因显著富集于核糖体、细胞周期以及癌症信号通路中。CLEC4M与癌症信号通路中的CCNB1、CCND3、APAF1、STEAP3、RRM2B、FAS这些基因相互作用共同促进脑星形胶质母细胞瘤增殖和迁移(P<0.05~P<0.01)。结论:CLEC4M促进脑星形胶质母细胞瘤的增殖和迁移。

Objective:To investigate the effect of CLEC4M on brain astroblastoma proliferation and migration,as well as the mechanism behind it.Methods:Brain astroblastoma cell lines overexpressing and interfering with CLEC4M were constructed by lentivirus transfection.Real-time quantitative PCR was used to determine the level of CLEC4M expression.The effects of CLEC4M on the proliferation and migration of brain astroblastoma were studied using the CCK-8 test and the cell scratch assay,and the effect of CLEC4M on the cell cycle of brain astroblastoma was studied using flow cytometry.Transcriptome sequencing was used to identify genes in cancer signaling pathways regulated by CLEC4M.Results:Brain astroblastoma cell lines overexpressing and interfering with CLEC4M were successfully constructed,and the overexpression and interference effect were significant(P<0.01).CLEC4M increased astroblastoma proliferation and migration,and it mostly affected the G0/G1 and S phases of the cell cycle to regulate cell cycle progression(P<0.05 to P<0.01).In addition,overexpression of CLEC4M in brain astroblastoma also modified the expression levels of 5857 genes,with 2897 genes highly up-regulated and 2960 genes significantly down-regulated,according to transcriptome sequencing studies.The differentially expressed genes were largely enriched in ribosomes,cell cycle,and cancer signaling pathways,according to KEGG analysis.CLEC4M interacted with CCNB1,CCND3,APAF1,STEAP3,RRM2B and FAS genes in cancer signaling pathways(P<0.05 to P<0.01).Conclusions:CLEC4M can increase the proliferation and migration of brain astroblastoma.