RNF213基因对急性髓系白血病THP-1细胞增殖及凋亡的影响  

The Effects of RNF2I3 on the Proliferation and Apoptosis of Acute Myeloid Leukemia THP-1 Cells

作　　者：施晓琦 张平平 官亚宁 杜作晨 陈艳 黄佩 何志旭 SHI Xiao-Qi;ZHANG Ping-Ping;GUAN Ya-Ning;DU Zuo-Chen;CHEN Yan;HUANG Pei;HE Zhi-Xu(Department of Pediatrics,The Affiliated Hospital of Zunyi Medical University,Department of Pediatrics,Guizhou Children's Hospital,Collaborative Innovation Center for Tissue Injury Repair and Regenerative Medicine of Zumyi Medical University,Zunyi 563000,Guizhou Province,China)

机构地区：[1]遵义医科大学附属医院小儿内科,贵州省儿童医院小儿内科,遵义医科大学组织损伤修复与再生医学省部共建协同创新中心,贵州遵义563000

出　　处：《中国实验血液学杂志》2024年第5期1365-1371,共7页Journal of Experimental Hematology

基　　金：贵州省科技计划项目(黔科合平台人才-CXTD[2021]010、ZK-2022-YB641);省部共建协同创新中心项目(教科技厅函[2020]39号);遵义市科技计划项目(遵市科人平台[2023]6号、遵市科合HZ字(2021)186号)。

摘　　要：目的:发掘RNF213基因与急性髓系白血病(AML)的关联,并探究RNF213对AML细胞系THP-1细胞增殖及凋亡的影响。方法:通过GEPIA数据库分析RNF213基因在AML中的表达与预后的关系。收集2017年1月至2022年1月于遵义医科大学附属医院就诊的AML患者30例以及血液系统非肿瘤患者21例,采用RT-qPCR和Western blot法分别检测RNF213 mRNA及蛋白表达水平,Kaplan-Meier法对AML患者进行生存分析,同时检测AML细胞株(THP-1、OCI-AML_(2))中RNF213 mRNA及蛋白表达水平。采用CRISPR-Cas9技术敲减THP-1细胞中RNF213基因,流式细胞术检测细胞凋亡率,CCK-8和集落形成实验检测细胞增殖情况,Western blot检测凋亡相关蛋白Cleaved-Caspase 3的表达水平。结果:AML患者的RNF213基因表达水平明显高于对照组,且RNF213高表达患者的预后更差,THP-1细胞中RNF213蛋白表达水平也高于对照组。敲减THP-1细胞RNF213基因后,细胞增殖明显降低,细胞凋亡率及凋亡相关蛋白Cleaved-Caspase3的表达较对照组明显增加。结论:在AML患者中RNF213高表达,且高表达患者预后更差。RNF213基因影响AML细胞增殖及凋亡,可能是AML预后相关的标志物和潜在的治疗靶点。Objective:To discover the relationship between the RNF213 gene and acute myeloid leukemia(AML),and explore the effct of RNF213 on the proliferation and apoptosis of THP-1 cells.Methods:Analyze the expression of RNF213 gene in AML and its relationship with prognosis through the GEPIA database.Collecting 30 AML patients and non-tumor hematological patients who went to the A filiated Hospital of Zunyi Medical University from January 2017 to January 2022.RT-qPCR and Western blot were used to detect the expression levels of RNF213 mRNA and protein.Perform survival of patients was analysed by Kaplan-Meier.Meanwhile,the expression levels of RNF213 mRNA and protein were detected in AML cell lines(THP-1,OCI-AML_(2)).CRISPR-Cas9 was used to knockdown the RNF213 gene in THP-1 cells;flow cytometry was used to detect apoptosis rate of cell.CCK-8 and colony formation assay were used to detect cell proliferation.Western blot was used to detect the expression level of Cleaved-Caspase 3 protein.Results:Compared with the control group,the expression level of RNF213 in AML patients was significantly increased,and patients with high expression of RNF213 have a worse prgnosis.Higher expression level of RNF213 protein in THP-1 cells.After knocking down the RNF213 gene of THP-1 cells,cell proliferation was significantly reduced,and the apoptosis rate and expression of apoptosis related protein Cleared-Caspase3 were significantly increased.Conclusion:AML patients have high expression of RNF213,and the prognosis of high expression patients is poor.The RNF213 gene affects AML cell proliferation and apoptosis,and may be a prognostic marker and potential therapeutic target for AML.

关 键 词：RNF213基因 急性髓系白血病 预后 增殖 凋亡 

分 类 号：R733.71[医药卫生—肿瘤]