过表达CASP1诱导人急性髓系白血病细胞THP-1的G0/G1细胞周期阻滞和NLRP3炎性小体介导的焦亡

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中文摘要：目的:探讨半胱天冬酶1(Caspase1，CASP1)对人急性髓系白血病(acute myeloid leukemi-a，AML)细胞THP-1 的细胞周期和细胞焦亡的影响。方法:培养 THP-1 细胞，将细胞分为对照组(正常培养的THP-1 细胞)，pcDNA3。1-null 组(过表达 CASP1 的阴性对照，用 5。0μg/mL 的 pcDNA3。1-null质粒转染THP-1 细胞 24h)、pcDNA3。1-CASP1 组(过表达 CASP1，用 5。0μg/mL 的 pcDNA3。1-CASP1 质粒转染THP-1 细胞24h)。用CCK-8 法检测各组细胞的增殖活力。用流式细胞术检测各组细胞凋亡和周期的变化。用qRT-PCR 法检测细胞中 CASP1、白细胞介素(interleukin，IL)-1β，IL-18的mRNA表达水平。用Western blot法检测细胞增殖相关蛋白 Ki67、增殖细胞核抗原(proliferating cell nuclear antigen，PCNA)，细胞周期蛋白D1(cyclin D1)、NOD 样受体热蛋白结构域相关蛋白 3(Nod-like receptor heat protein domain associated protein 3，NLRP3)、凋亡相关斑点样蛋白((apoptosis-associated speck-like protein，ASC)、CASP1、切割半胱天冬酶1(cleaved-caspase 1，cleaved-CASP1)、IL-1β，IL-18、cleaved-Gasdermin D的相对表达水平。结果:与对照组比，pcDNA3。1-null 组的细胞增殖活力、细胞凋亡、细胞周期变化均无统计学意义(P>0。05)。与对照组比，pcDNA3。1-CASP1 组的细胞凋亡变化无统计学意义(P>0。05)，细胞的增殖活力减少，G0/G1 细胞周期被阻滞，Ki67、PCNA、cyclin D1 的相对表达水平均减少(P<0。05)，NLRP3、ASC、CASP1、cleaved-CASP1、IL-1β，IL-18、cleaved-Gasdermin D 的相对表达水平均增加(P<0。05)。与pcDNA3。1-null组比，pcDNA3。1-CASP1 组的细胞凋亡变化无统计学意义(P>0。05)，细胞的增殖活力减少，G0/G1 细胞周期被阻滞，Ki67、PCNA、cyclin D1 的相对表达水平均减少(P<0。05)，NLRP3、ASC、CASP1、cleaved-CASP1、IL-1β，IL-18、cleaved-Gasdermin D(30 kDA)的相对表达水平均增加(P<0。05)。结论:过表达 CASP1 诱导 AML 细胞 THP-1 的 G0/G1 细胞周期阻滞和NLRP3 炎性小体介导的焦亡。

外文标题：Overexpression of Caspase1 Induces G0/G1 Cell Cycle Arrest and NLRP3 Inflammasome-Mediated Pyroptosis in Human Acute Myeloid Leukemia Cells THP-1

外文摘要：Objective:To investigate the effects of caspase1(CASP1)on the cell cycle and pyroptosis of human acute myeloid leukemia(AML)cells THP-1.Methods:THP-1 cells were cultured and divided in-to the control group(normal THP-1 cells),pcDNA3.1-null group(negative control for overexpression of CASP1,transfected with 5.0 μg/mL pcDNA3.1-null plasmid into THP-1 cells for 24 h),and pcDNA3.1-CASP1 group(overexpression of CASP1,transfected with 5.0 μg/ml pcDNA3.1-CASP1 plasmid into THP-1 cells for 24 h).Cell proliferation activity was detected by CCK-8 assay.Apoptosis and cell cycle were de-tected by flow cytometry.CASP1,interleukin(IL)-1β,and IL-18 mRNA expression levels were detected by qRT-PCR.Cell proliferation-related proteins Ki67,proliferating cell nuclear antigen(PCNA),cyclin D1,NOD-like receptor(NLR)-like receptor pyrin domain-containing protein 3(NLRP3),apoptosis-asso-ciated speck-like protein(ASC),CASP1,cleaved-caspase 1(cleaved-CASP1),IL-1β,IL-18,and cleaved-Gasdermin D were detected by Western blot.Results:Compared with the control group,there were no significant differences in cell proliferation activity,apoptosis,and cell cycle changes in the pcDNA3.1-null group(P>0.05).Compared with the control group,cell proliferation activity was reduced,G0/G1 cell cycle was blocked,and the relative expression levels of Ki67,PCNA,and cyclin D1 were decreased(P<0.05)in the pcDNA3.1-CASP1 group.The relative expression levels of NLRP3,ASC,CASP1,cleaved-CASP1,IL-1β,IL-18,and cleaved-Gasdermin D were increased(P<0.05).Compared with the pcDNA3.1-null group,there were no significant differences in apoptosis in the pcDNA3.1-CASP1 group(P>0.05).However,cell proliferation activity was reduced,G0/G1 cell cycle was blocked,and the relative expression levels of Ki67,PCNA,and cyclin D1 were decreased(P<0.05).The relative expression levels of NLRP3,ASC,CASP1,cleaved-CASP1,IL-1β,IL-18,and cleaved-Gasdermin D were increased(P<0.05).Con-clusion:Overexpression of CASP1 induces G0/G1 cell cycle arrest and NLRP3 inflammasome-mediated py-roptosis in AML cells THP-1.

外文关键词：

Human acute myeloid leukemia cellsCaspase 1Cell cyclePyroptosis

作者：

艾克拜尔·阿布都热衣木、徐丽、阿孜古丽·麦麦提、阿依姆妮萨·阿卜杜热合曼、帕提古力·苏力坦

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作者单位：

新疆维吾尔自治区喀什地区第一人民医院血液内科, 新疆喀什 844000

关键词：

人急性髓系白血病细胞半胱天冬酶1 细胞周期细胞焦亡

基金：

新疆少数民族科技人才特殊培养计划

项目编号：

2018D03020

出版年：

2024

DOI：

10.3969/j.issn.1006-6233.2024.02.06

河北医学

河北省医学会

河北医学

CSTPCD

影响因子：1.915

ISSN：1006-6233

年,卷(期)：2024.30(2)

参考文献量11