整合多数据库分析PM20D1基因与弥漫大B细胞淋巴瘤缺氧相关性及预后的关系  

The relationship between PM20D1 gene and hypoxia and prognosis in diffuse large B-cell lymphoma was analyzed by integrating multiple databases

作　　者：张园园 潘静 李光明 焦霞霞 赵文理 ZHANG Yuanyuan;PAN Jing;LI Guangming;JIAO Xiaxia;ZHAO Wenli(Medical College,Anhui University of Science and Technology,Huainan 232000,Anhui,China;Department of Hematology,Shanghai Fengxian District Central Hospital,Shanghai 201499;Hubei University of Medicine,Shiyan 442000,Hubei,China)

机构地区：[1]安徽理工大学医学院,安徽淮南232000 [2]上海市奉贤区中心医院血液内科,上海201499 [3]湖北医药学院,湖北十堰442000

出　　处：《中国肿瘤生物治疗杂志》2023年第5期401-411,共11页Chinese Journal of Cancer Biotherapy

基　　金：上海市卫生健康委员会重点科研项目(No.201640028)。

摘　　要：目的:探讨肽酶M20结构域1(peptidase M20 domain containing 1,PM20D1)在人弥漫大B细胞淋巴瘤(diffuse large B-cell lymphomas,DLBCL)细胞中的表达及其与缺氧相关性和预后的关系。方法:通过GDC、TCGA、GTEx公共数据库分析PM20D1表达对DLBCL细胞增殖、迁移和凋亡的影响及其与患者预后的关系。通过对不同分组患者的富集分析及PM20D1与CD274相关性分析验证PM20D1是否为DLBCL的缺氧相关基因;采用ChEA、ENCODE和hTFtarget数据库分析上游调控PM20D1表达的转录因子(TF)和miRNA,以及差异表达PM20D1与化疗药物敏感性的关系。采用WB法检测PM20D1在正常淋巴细胞和DLBCL细胞中的表达水平,通过设计PM20D1的siRNA序列敲减目的基因,并采用qPCR和WB法检测验证SUDHL2和SUDHL10细胞中PM20D1的敲减效率,采用CCK-8法和Transwell实验分别检测敲减PM20D1对细胞增殖和迁移能力的影响,流式细胞术检测细胞凋亡水平。结果:PM20D1在DLBCL组织中高表达且患者预后差(P<0.05或P<0.01);富集分析显示,PM20D1高表达组与ssGSEA高分组主要涉及细胞电耦合通讯、甘油三酯代谢过程调节和细胞质翻译起始复合物过程,且PM20D1表达与免疫检查点CD274表达呈正相关(P<0.01,r=0.757)。在SUDHL2和SUDHL10细胞中敲减PM20D1后,细胞的增殖和迁移均显著降低(均P<0.05),细胞凋亡明显增加(P<0.05)。结论:PM20D1基因在DLBCL组织和细胞中高表达且与患者预后密切相关;PM20D1可能通过促进DLBCL细胞的增殖、迁移并抑制凋亡,从而促进DLBCL的发生发展。Objective:To investigate the expression of peptidase M20 domain 1(PM20D1)in human diffuse large B-cell lymphoma(DLBCL)cells and its relationship with hypoxia and prognosis.Methods:The effects of PM20D1 expression on proliferation,migration and apoptosis of DLBCL cells and its relationship with patients'prognosis were analyzed through GDC,TCGA and GTEx public databases.The enrichment analysis of patients in different groups and the correlation analysis between PM20D1 and CD274 were used to validate whether PM20D1 was the hypoxia-related gene of DLBCL.ChEA,ENCODE and hTFtarget databases were used to analyze the transcription factors(TFs)and miRNAs that upstream regulate PM20D1 expression and the relationship between differential expression of PM20D1 and chemotherapeutic drug sensitivity.The expression level of PM20D1 in normal lymphocytes and DLBCL cells was detected by WB method,the target gene was reduced by the siRNA sequence of PM20D1,and the knockdown efficiency of PM20D1 in SUDHL2 and SUDHL10 cells was verified by qPCR and WB methods.The effect of PM20D1 on cell proliferation and migration ability after knockdown was detected by CCK-8 method and Transwell assays,respectively,and the apoptosis level was detected by flow cytometry.Results:PM20D1 was highly expressed in DLBCL tissues and was correlated with a poor prognosis(P<0.05 or P<0.01).Enrichment analysis showed that the PM20D1 high expression group and high ssGSEA score group were mainly involved in cellular electrocoupling communication,triglyceride metabolism process regulation and cytoplasmic translation initiation complex process,and PM20D1 expression was positively correlated with CD274 expression at immune checkpoint(P<0.01,r=0.757).After knocking down PM20D1 in SUDHL2 and SUDHL10 cells,the proliferation and migration of cells decreased significantly(all P<0.05),and apoptosis increased significantly(P<0.05).Conclusion:PM20D1 gene was highly expressed in DLBCL tissues and cells,and in closely related to patient prognosis.PM20D1 may promote the occurr

关 键 词：肽酶M20结构域1(PM20D1) 弥漫大B细胞淋巴瘤(DLBCL) 肿瘤缺氧 增殖 迁移 凋亡 

分 类 号：R733.4[医药卫生—肿瘤] R730.7[医药卫生—临床医学]