GOT2调控急性髓系白血病细胞的天冬氨酸含量和存活  

GOT2 regulates aspartate levels and viability in acute myeloid leukemia cells

作　　者：刘译阳 王芳[1] 余佳[1] 李卫倩 LIU Yiyang;WANG Fang;YU Jia;LI Weiqian(State Key Laboratory of Common Mechanism Research for Major Diseases,Institute of Basic Medical Sciences CAMS,School of Basic Medicine PUMC,Bejing 100005,China)

机构地区：[1]中国医学科学院基础医学研究所,北京协和医学院基础学院,医学分子生物学国家重点实验室,北京100005

出　　处：《基础医学与临床》2025年第5期622-626,共5页Basic and Clinical Medicine

基　　金：国家自然科学青年科学基金(82300140)。

摘　　要：目的探讨谷氨酸-草酰乙酸转氨酶2基因(GOT2)在急性髓系白血病细胞中的功能。方法构建针对GOT2的shRNA,使用shRNA在人单核细胞白血病细胞系THP-1中抑制GOT2基因的表达。通过RT-qPCR检测GOT2 mRNA表达水平,Western blot检测GOT2蛋白表达水平的变化。采用比色法检测细胞内游离氨基酸含量变化。检测细胞GOT2表达缺失对THP-1细胞功能的影响,包括:CCK8法评估细胞增殖情况;流式细胞测量术分析细胞凋亡水平。结果使用GOT2 shRNA干扰后,THP-1细胞GOT2的mRNA及蛋白表达水平均显著降低(P<0.05)。与对照组相比,敲低GOT2不影响细胞内谷氨酸含量(P>0.05),但会显著下调天冬氨酸含量(P<0.05);敲低GOT2显著降低了THP-1细胞的活性,使细胞增殖受阻(P<0.05)、凋亡增加(P<0.05)。结论下调GOT2可显著降低急性髓系白血病细胞系内天冬氨酸水平,抑制细胞生长并诱导细胞凋亡,提示GOT2在急性髓系白血病的氨基酸代谢调控中可能发挥关键作用。Objective To investigate the function of the glutamic-oxaloacetic transaminase 2(GOT2)gene in acute myeloid leukemia cells.Methods ShRNAs targeting at GOT2 were constructed to suppress GOT2 expression in the THP-1 human monocytic leukemia cell line.GOT2 mRNA level was measured by RT-qPCR,and GOT2 protein expression was measured by Western blot.Intracellular free amino acid level was quantified using colorimetric assays.The impact of GOT2 knockdown in THP-1 cell function was evaluated through cell viability(CCK-8 assay)and apoptosis(flow cytometry).Results After GOT2 knockdown,both GOT2 mRNA and protein were significantly decreased in THP-1 cells(P<0.05).Compared to the control group,GOT2 knockdown did not affect intracellular glutamate levels(P>0.05),but led to a marked decrease in aspartate level(P<0.05).GOT2 knockdown significantly impaired THP-1 cell viability,inhibited cell proliferation(P<0.05)and promoted apoptosis(P<0.05).Conclusions Down regulation of GOT2 significantly decreases intracellular aspartate level in acute myeloid leukemia cells,impairs cellular viability and induces apoptosis,which suggests that GOT2 may play a key role in the regulation of amino acid metabolism in acute myeloid leukemia.

关 键 词：GOT2 急性髓系白血病 天冬氨酸 

分 类 号：R733.71[医药卫生—肿瘤]