circPCMTD1对AML细胞恶性行为影响及机制  

Effect of circPCMTD1 on malignant behavior of acute myeloid leukemia cells and its mechanism

作　　者：陆静忠 方永修 黄美琴 王小红 LU Jingzhong;FANG Yongxiu;HUANG Meiqin;WANG Xiaohong(Department of Hematology,Chongming Hospital Affiliated to Shanghai University of Health Sciences,Shanghai 202150,China)

机构地区：[1]上海健康医学院附属崇明医院血液内科,上海202150 [2]上海市崇明区精神卫生中心精神科 [3]上海健康医学院附属崇明医院中医内科 [4]上海健康医学院附属崇明医院门急诊办公室

出　　处：《青岛大学学报（医学版）》2024年第5期663-668,共6页Journal of Qingdao University(Medical Sciences)

基　　金：上海市崇明区“可持续发展科技创新行动计划”项目(CKY2022-04)。

摘　　要：目的探讨circPCMTD1对急性髓系白血病(AML)细胞增殖、迁移、侵袭和凋亡的影响及其分子机制。方法体外培养人骨髓基质细胞系HS-5与AML细胞系HL-60、THP-1、U-937和Kasumi-1。将THP-1细胞随机分为NC组、si-NC组、si-circPCMTD1组、miR-328 NC组、miR-328-3p组和si-circPCMTD1+anti-miR-NC/anti-miR-328-3p组。采用实时荧光定量PCR(RT-qPCR)和Western blot法分别检测circPCMTD1、miR-328-3p表达量和相关蛋白的表达量;采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯四唑溴化铵(MTT)法、Transwell实验和流式细胞术分别检测细胞增殖、迁移、侵袭和凋亡;并通过双荧光素酶报告实验评估circPCMTD1和miR-328-3p的关系。结果与HS-5细胞比较,circPCMTD1在AML细胞中高表达(F=66.258,P<0.05),miR-328-3p则相反(F=101.145,P<0.05)。下调circPCMTD1抑制了细胞的增殖、迁移、侵袭及其相关蛋白表达,并诱导了细胞的凋亡(F=38.952~290.338,P<0.05)。上调miR-328-3p可削弱WT-circPCMTD1的荧光素酶活性,且下调miR-328-3p可减弱circPCMTD1下调对细胞增殖的抑制作用(F=42.655,P<0.05)。结论沉默circPCMTD1可通过靶向miR-328-3p抑制AML细胞的生长。Objective To explore the effects of circPCMTD1 on the proliferation,migration,invasion,and apoptosis of acute myeloid leukemia(AML)cells and the underlying molecular mechanism.Methods Human bone marrow stromal cell line HS-5 and AML cell lines HL-60,THP-1,U-937,and Kasumi-1 were cultured in vitro.THP-1 cells were randomly divided into NC group,si-NC group,si-circPCMTD1 group,miR-328 NC group,miR-328-3p group,si-circPCMTD1+anti-miR-328 NC group,and si-circPCMTD1+anti-miR-328-3p group.Real-time fluorescence quantitative PCR and Western blot were used to mea-sure the expression of circPCMTD1,miR-328-3p,and related proteins.Cell proliferation,migration,invasion,and apoptosis were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT),Transwell and flow cytometry methods.The relationship between circPCMTD1 and miR-328-3p was analyzed by dual luciferase reporting assay.Results Compared with HS-5 cells,circPCMTD1 was highly expressed in AML cells(F=66.258,P<0.05),whereas the opposite was true for miR-328-3p(F=101.145,P<0.05).Silencing circPCMTD1 inhibited the proliferation,migration,and invasion of cells and the expression of related proteins,and induced cell apoptosis(F=38.952-290.338,P<0.05).Upregulation of miR-328-3p impaired the luciferase activity of WT-circPCMTD1,and downregulation of miR-328-3p attenuated the inhibitory effect of circPCMTD1 downregulation on cell proliferation(F=42.655,P<0.05).Conclusion Silencing circPCMTD1 can repress THP-1 cell growth through targeting miR-328-3p.

关 键 词：白血病 髓样 急性 RNA 环状 微RNAS 细胞增殖 细胞运动 肿瘤浸润 细胞凋亡 

分 类 号：R733.72[医药卫生—肿瘤]