神经节苷脂GD2可作为肺癌干细胞一个潜在的候选标志物  

Ganglioside GD2 is a potential candidate marker for lung cancer stem cells

作　　者：李珍 段昭君 罗云萍[1] LI Zhen;DUAN Zhaojun;LUO Yunping(Department of Immunology,Institute of Basic Medical Sciences CAMS,School of Basic Medicine PUMC,Beijing 100005,China)

机构地区：[1]中国医学科学院基础医学研究所、北京协和医学院基础学院免疫学系,北京100005

出　　处：《基础医学与临床》2023年第5期777-784,共8页Basic and Clinical Medicine

基　　金：中央高校基本科研业务费专项资金(3332022181)。

摘　　要：目的探索神经节苷脂GD2作为肺癌干细胞(LCSCs)标志物的可行性。方法用成球培养的方式在体外富集肺癌细胞系的肿瘤干细胞(CSCs);用实时定量PCR和蛋白质免疫印迹法检测LCSCs中干性相关基因以及GD3合成酶(GD3S)的表达;用流式细胞测量术检测肺腺癌细胞系A549中GD2及其他干性表型的表达;用CCK8法检测细胞增殖;用Transwell小室法和细胞划痕愈合实验检测细胞的体外侵袭和迁移;用肺癌移植瘤模型比较细胞系A549中不同亚群的致瘤性。结果体外成球培养能够成功富集到A549细胞的肿瘤干细胞,贴壁培养时,GD2+细胞的比例为0.57%,成球培养后GD2+细胞比例能够上升至20.4%。与GD2-A549细胞相比,GD2+A549细胞的体外增殖(P<0.05)、侵袭(P<0.05)和迁移能力(P<0.01)均显著提高。GD2与另外两种肺癌干细胞表型之间有密切联系,其表达水平与CD44(P<0.01)及EpCAM(P<0.001)的表达呈正相关。体内的结果表明,相比于GD2-A549细胞,接种GD2+A549细胞后肿瘤生长更快(P<0.001),恶性程度更高(P<0.01)。结论神经节苷脂GD2可作为LCSCs的潜在标志物以识别肿瘤干细胞,将GD2作为肺癌免疫治疗的靶点提供了新的思路。Objective To explore the feasibility of ganglioside GD2 as a marker for lung cancer stem cells(LCSCs).Methods Cancer stem cells(CSCs)of lung cancer cell lines were enriched by spheroid culturing in vitro;The expression of stemness-related genes and GD3 synthase(GD3S)in LCSCs were detected by RT-qPCR and Western blot;The expression of GD2 and other stemness phenotypes in lung cancer cell line A549 were detected by flow cytometry;Cell proliferation was detected by CCK8 assay;Cell invasion and migration in vitro were examined by Transwell and wound healing assays;The oncogenicity of different subpopulations in cell line A549 was compared with tumor xenograft model.Results CSCs of A549 cell were successfully enriched by and the percentage of GD2+cells was 0.57%in adherent cell,but the percentage of GD2+cells was able to increase to 20.4%after spheroid culturing.In comparison with GD2-A549 cells,GD2+A549 cells showed significantly enhanced cell proliferation(P<0.05),invasion(P<0.05)and migration ability(P<0.01)in vitro.There was a close association between GD2 and two other LCSCs phenotypes,and the expression level of GD2 was positively correlated with the expression of CD44(P<0.01)and EpCAM(P<0.001).The results in vivo indicated that tumors formed by GD2+A549 cells grew faster(P<0.001)and were more malignant(P<0.01)as compared to GD2-A549 cells.Conclusions Ganglioside GD2 is a potential marker for LCSCs to identify cancer stem cells,providing a novel strategy of GD2-targeted lung cancer immunothearapy.

关 键 词：肺癌干细胞 神经节苷脂GD2 肿瘤相关抗原 免疫治疗 

分 类 号：R734.2[医药卫生—肿瘤]