Objective: To discover the relationship between the RNF213 gene and acute myeloid leukemia (AML), and explore the effect 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 Affiliated 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-AML2 ). 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.
题目: RNF213 基因对急性髓系白血病THP-1细胞增殖及凋亡的影响.
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-AML2 )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。.
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