癌-睾丸抗原CT63在慢性髓系白血病中的作用及其机制  

Function and mechanism of cancer-testis antigen CT63 in chronic myeloid leukemia

作　　者：孔汝心 周亚群 魏婷宜 雷鸣 KONG Ruxin;ZHOU Yaqun;WEI Tingyi;LEI Ming(Shanghai Institute of Precision Medicine,Shanghai Ninth People's Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200125,China)

机构地区：[1]上海交通大学医学院附属第九人民医院精准医学研究院,上海200125

出　　处：《上海交通大学学报（医学版）》2024年第11期1347-1358,共12页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家重点研发计划(2018YFA0107004);国家自然科学基金(32300609)。

摘　　要：目的·研究癌-睾丸抗原(cancer-testis antigen,CTA)家族成员CT63对慢性髓系白血病细胞增殖、分化和成瘤方面的影响,阐述其分子机制。方法·采用生物信息学方法分析癌症基因组图谱(TCGA)数据库中髓系白血病患者的CT63表达差异及其对疾病预后的影响;构建CT63敲降的人慢性髓系白血病K562细胞系,通过实时荧光定量PCR和蛋白质印迹法确定CT63的敲降效果;通过细胞实时成像和CCK-8实验评估CT63对慢性髓系白血病细胞增殖能力的影响;进行小鼠皮下成瘤实验,探究CT63在体内环境下对慢性髓系白血病肿瘤发生、生长和细胞分化的影响;通过佛波酯诱导的K562细胞向单核/巨噬细胞的分化实验,探究CT63对慢性髓系白血病细胞分化能力的影响;检测敲降CT63后细胞内线粒体功能的变化。结果·Kaplan-Meier生存曲线分析显示,CT63差异表达与髓系白血病患者的预后显著相关;敲降CT63抑制K562细胞的增殖和成瘤;敲降CT63促进K562细胞在体内和体外的分化;敲降CT63抑制K562细胞中线粒体呼吸链复合物Ⅳ的活性,并导致包括细胞色素C氧化酶Ⅳ(COXⅣ)、丙酮酸脱氢酶、琥珀酸脱氢酶A(SDHA)和电压依赖性阴离子通道(VDAC)的线粒体相关标志物表达下降,影响K562细胞的线粒体代谢活性。结论·CT63的表达水平与髓系白血病患者的预后相关;CT63通过维持线粒体的代谢活性,促进K562细胞的增殖和体内成瘤;CT63是维持慢性髓系白血病细胞自我更新和分化的重要分子开关。Objective·To explore the effects of the cancer-testis antigen(CTA)family member CT63 on proliferation,differentiation,and tumorigenicity of chronic myeloid leukemia(CML)cells,and uncover the underlying molecular mechanisms.Methods·The link between CT63 expression and the prognosis of myeloid leukemia patients was analyzed using bioinformatics methods(TCGA database).A K562 cell line with CT63 knockdown was established.The knockdown efficiency of CT63 was confirmed by qRT-PCR and Western blotting.Live-cell imaging and CCK-8 methods were adopted to evaluate the inhibitory effect of CT63 knockdown in CML cells.A subcutaneous tumorigenesis assay in nude mice was conducted to examine the effects of CT63 on tumorigenesis,tumor growth,and differentiation of K562 cells in vivo.Phorbol 12-myristate 13-acetate(PMA)-induced monocyte/macrophage differentiation experiment was carried out to investigate the role of CT63 in the differentiation of K562 cells in vitro.Mitochondrial function was assessed to determine the impact of CT63 on CML cells both in vivo and in vitro.Results·The Kaplan-Meier survival curve indicated that low expression levels of CT63 were correlated with longer survival in patients with myeloid leukemia.Down-regulation of CT63 in K562 cells inhibited proliferation and promoted differentiation.Live-cell imaging and CCK-8 assays displayed that knockdown of CT63 inhibited cell proliferation and extended cell doubling time in K562 cells.In the subcutaneous xenotransplantation model,down-regulation of CT63 inhibited tumor growth in nude mice.K562 cells expressing lower levels of CT63 were more prone to differentiate into monocyte/macrophage both in vivo and in vitro under PMA exposure condition.Knockdown of CT63 suppressed the activity of mitochondrial respiratory chain complexⅣ.This led to decreased expression of mitochondrial markers,including cytochrome C oxidaseⅣ(COXⅣ),pyruvate dehydrogenase,succinate dehydrogenase A(SDHA),and voltage-dependent anion channel(VDAC),thus affecting the mitochondrial met

关 键 词：癌-睾丸抗原63 慢性髓系白血病 细胞增殖 细胞分化 线粒体 

分 类 号：R733.72[医药卫生—肿瘤]