[Ku80抑制对t -急性淋巴母细胞白血病Jurkat细胞化疗敏感性的影响]。

Q4 Medicine
Zhuo-Yi Fan, Ai-Bin Liang
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引用次数: 0

摘要

目的:探讨Ku80抑制对t -急性淋巴细胞白血病(T-ALL) Jurkat细胞化疗敏感性的影响,并探讨其可能的机制。方法:分别采用RT-qPCR和Western blot检测6种血液恶性细胞株中Ku80的转录和表达水平。重组shKu80慢病毒载体转染Jurkat细胞后,用Western blot检测Ku80的表达。通过CCK-8检测Ku80抑制后Jurkat细胞的增殖能力。通过软琼脂法、流式细胞术和Western blot分别研究Ku80沉默和依托opo苷(VP16)共处理4小时后Jurkat细胞的集落形成能力、凋亡和γ - h2ax (DNA损伤蛋白标志物)的表达。结果:6种血液恶性细胞系中,Ku80 mRNA水平和蛋白表达均在Jurkat中最高。转染相关质粒后,Ku80在Jurkat细胞中的表达成功下调。Ku80抑制后细胞增殖能力显著降低(P < 0.05)。Ku80抑制后,Jurkat细胞的集落形成能力明显降低,细胞凋亡和γ - h2ax表达增加。结论:靶向沉默Ku80可增强Jurkat细胞中VP16的敏感性,这可能与DNA损伤积累水平升高有关。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Ku80 Inhibition Affects the Chemotherapeutic Sensitivity of T-Acute Lymphoblastic Leukemia Cell Line Jurkat].

Objective: To investigate the influence of Ku80 inhibition on the chemotherapeutic sensitivity of the T-acute lymphoblastic leukemia(T-ALL) cell line Jurkat, and to explore the potential mechanism.

Methods: The transcription and expression level of Ku80 in 6 hematological malignant cell lines were detected by RT-qPCR and Western blot, respectively. The expression of Ku80 in Jurkat cells was detected by Western blot after transfection with the recombinant shKu80 lentiviral vector. The proliferation capacity of Jurkat cells was explored by CCK-8 after Ku80 inhibition. The colony formation ability, apoptosis, and γH2AX(a protein marker of DNA damage) expression in Jurkat cells were investigated after Ku80 silencing and co-treated with etoposide(VP16) for 4 hours through soft agar assay, flow cytometry and Western blot, respectively.

Results: The mRNA level and protein expression of Ku80 were both highest in Jurkat among 6 hematological malignant cell lines. Ku80 expression was successfully down regulated in Jurkat cells after relative plasmid transfected. The proliferative ability of cells was significantly decreased after Ku80 inhibition(P < 0.05). The colony formation capacity of Jurkat cells was obviously reduced and the cells apoptosis and γH2AX expression were increased after Ku80 inhibition, with or without VP16 incubation.

Conclusion: Targeted silencing of Ku80 could enhance the sensitivity of VP16 in Jurkat cells, which might be associated with the elevated level of DNA damage accumulation.

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来源期刊
中国实验血液学杂志
中国实验血液学杂志 Medicine-Medicine (all)
CiteScore
0.40
自引率
0.00%
发文量
7331
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