SFXN2介导的线粒体自噬参与弥漫性大B细胞淋巴瘤的利妥昔单抗耐药机制研究  

Study on the role of SFXN2-mediated mitophagy in the mechanism of rituximab resistance in diffuse large B-cell lymphoma

作　　者：胡陶 杜春燕 李正斯 HU Tao;DU Chunyan;LI Zhengsi(Department of Pediatrics,Mianyang Central Hospital,Mianyang Hospital Affiliated to School of Medicine,University of Electronic Science and Technology of China,Mianyang 621000,Sichuan,China)

机构地区：[1]绵阳市中心医院电子科技大学医学院附属绵阳医院儿科,四川绵阳621000

出　　处：《中南医学科学杂志》2025年第2期211-216,共6页Medical Science Journal of Central South China

基　　金：四川省医学科研课题(s19068);绵阳市卫健委课题(201913)。

摘　　要：目的 探讨铁氧蛋白2(SFXN2)介导的线粒体自噬参与弥漫性大B细胞淋巴瘤(DLBCL)的利妥昔单抗耐药机制。方法 将DLBCL细胞系(OCI-LY1和RL)和利妥昔单抗耐药细胞系(RL-4RH)实验分为空载体(Vector)/OCI-LY1组、SFXN2/OCI-LY1组、短发夹SFXN2(shSFXN2)/RL组及其阴性对照(shNC)/RL组、shNC/RL-4RH组、shSFXN2/RL-4RH组、线粒体自噬阻断剂氯喹(CQ)/RL-4RH组、CQ+shSFXN2/RL-4RH组。比较各组SFXN2蛋白表达水平。采用CCK-8、FACS分析各组细胞活力及凋亡情况。激光共聚焦显微镜下观察RL-4RH细胞在敲低SFXN2后的自噬通量。裸鼠实验分为RL组(接种RL肿瘤细胞)、RL-4RH组(接种RL-4RH肿瘤细胞)和RL-4RH+shSFXN2组(接种转染shSFXN2的RL-4RH肿瘤细胞),各组动物给予利妥昔单抗干预后,采用免疫组织化学染色分析各组肿瘤组织Ki67和SFXN2表达情况。结果 与shNC/RL组相比,shSFXN2/RL组SFXN2蛋白表达水平、细胞活力降低(P<0.05),细胞凋亡率增加(P<0.05);与Vector/OCI-LY1组相比,SFXN2/OCI-LY1组SFXN2蛋白表达水平、细胞活力升高(P<0.05)。与shNC/RL-4RH组相比,shSFXN2/RL-4RH组自噬体数量减少(P<0.05),自噬空泡、自溶体数量增加(P<0.05)。与shSFXN2/RL-4RH组相比,CQ+shSFXN2/RL-4RH组自噬体数量增加(P<0.05),自溶体数量减少(P<0.05)。与RL组相比,RL-4RH组荷瘤小鼠肿瘤体积增加(P<0.05),肿瘤组织中SFXN2、Ki67表达水平上调(P<0.05);RL-4RH+shSFXN2组逆转了RL-4RH组荷瘤小鼠中的利妥昔单抗耐药性效应(P<0.05)。结论SFXN2敲低可能通过诱导线粒体自噬的过度激活,逆转利妥昔单抗耐药性。Aim To explore the mechanism of rituximab resistance in diffuse large B-cell lymphoma(DLBCL) through mitochondrial autophagy mediated by sideroflexin 2(SFXN2).Methods The DLBCL cell lines(OCI-LY1 and RL) and rituximab resistant cell lines(RL-4RH) were divided into Vector/OCI-LY1 group,SFXN2/OCI-LY1 group,short hairpin SFXN2(shSFXN2)/RL group and its negative control(shNC)/RL group,shNC/RL-4RH group,shSFXN2/RL-4RH group,mitochondrial autophagy inhibitor Chloroquine(CQ)/RL-4RH group,and CQ+shSFXN2/RL-4RH group in this study.The expression level of SFXN2 protein in each group was compared.CCK-8 and FACS were used to analyze cell viability and apoptosis in each group.Autophagy flux of RL-4RH cells after knocking down SFXN2 was observed under laser confocal microscopy.The nude mice were divided into RL group(inoculated with RL tumor cells),RL-4RH group(inoculated with RL-4RH tumor cells),and RL-4RH+shSFXN2 group(inoculated with shSFXN2 transfected RL-4RH tumor cells).After intervention with rituximab,the expression of Ki67 and SFXN2 in tumor tissues of each group was analyzed by immunohistochemical staining.Results Compared with the shNC/RL group,the shSFXN2/RL group showed a decrease in SFXN2 protein expression level and cell viability(P<0.05),but an increase in cell apoptosis rate(P<0.05).Compared with the Vector/OCI-LY1 group,the SFXN2 protein expression level and cell viability were increased in the SFXN2/OCI-LY1 group(P<0.05).Compared with the shNC/RL-4RH group,the shSFXN2/RL-4RH group showed a decrease in autophagosomes(P<0.05),but an increase in the number of autophagic vacuoles and autolysosomes(P<0.05).Compared with the shSFXN2/RL-4RH group,the CQ+shSFXN2/RL-4RH group showed an increase in autophagosomes(P<0.05) but a decrease in the number of autolysosomes(P<0.05).Compared with the RL group,the RL-4RH group showed an increase in tumor volume in tumor bearing mice(P<0.05),and upregulation of SFXN2 and Ki67 expression in tumor tissue(P<0.05);Knockdown of SFXN2 reversed the rituximab resistance effect in RL-4RH

关 键 词：铁氧蛋白2 线粒体 自噬 弥漫性大B细胞淋巴瘤 利妥昔单抗 耐药 

分 类 号：R73.3[医药卫生—肿瘤]