PDF(20555 KB)
PDF(20555 KB)
PDF(20555 KB)
敲低去泛素化酶USP13抑制K562细胞的增殖*
Knockdown of Deubiquitinase USP13 Inhibits the Proliferation of K562 Cells
({{custom_author.role_en}}), {{javascript:window.custom_author_en_index++;}}目的:研究去泛素化酶USP13对人慢性髓系白血病细胞系K562增殖和凋亡的影响,并进行初步的机制探究。方法:构建pLKO.1-shUSP13-GFP慢病毒干涉载体,慢病毒包装后感染并建立稳定敲低USP13的K562细胞株。免疫印迹检测K562细胞中USP13蛋白的敲低效率。流式细胞术分析敲低USP13对K562细胞增殖和凋亡的影响。免疫共沉淀和蛋白质泛素化实验探究USP13调控K562细胞的分子机制。结果:成功构建pLKO.1-shUSP13-GFP慢病毒干涉载体,同时利用慢病毒体系获得稳定敲低USP13的K562细胞株。流式细胞术结果显示,敲低USP13促进K562细胞凋亡、抑制细胞增殖。分子机制研究发现,敲低USP13通过增强c-Myc泛素化进而导致其蛋白质水平降低。结论:初步揭示了USP13调控K562细胞增殖和凋亡的分子机制,为治疗慢性髓系白血病提供了潜在的靶点。
Objective: Study the effect of deubiquitinating enzyme USP13 on the proliferation and apoptosis of human chronic myeloid leukemia K562 cells, and explore the underlying mechanism. Methods: Construction of the pLKO.1-shUSP13-GFP lentiviral interference vector and establishment of the USP13 knockdown K562 cell line using lentivirus. Western blot detected the USP13 knockdown efficiency in K562 cells. Flow cytometry analyzed the effect of USP13 knockdown on the proliferation and apoptosis of K562 cells. Co-immunoprecipitation and protein ubiquitination experiments explored the regulation mechanism of USP13 on K562 cells. Results: The pLKO.1-shUSP13-GFP vector was successfully constructed, and K562 cell line with stable knockdown of USP13 was obtained using the lentiviral system. Flow cytometry results showed that knocking down USP13 promoted K562 cell apoptosis and inhibited cell proliferation. Molecular mechanism studies found that knockdown of USP13 decreases c-Myc level by enhancing its ubiquitination. Conclusions: Data preliminarily revealed the molecular mechanism of USP13 regulating the proliferation and apoptosis of K562 cells, providing a potential target for the treatment of chronic myeloid leukemia.
人慢性髓系白血病 / K562细胞 / USP13 / 增殖和凋亡 / c-Myc {{custom_keyword}} /
Chronic myelogenous leukemia / K562 cells / USP13 / Proliferation and apoptosis / c-Myc {{custom_keyword}} /
| [1] |
: The outlook for patients with chronic myeloid leukemia (CML) has changed dramatically with the development of tyrosine kinase inhibitors (TKIs) with the current treatment goal for many patients being to obtain a durable deep molecular remission, discontinue TKI therapy, and remain treatment free.: In this article, the authors review the data from the major TKI discontinuation studies, explore potential predictors of discontinuation outcome and look at possible mechanisms to explain the variable outcomes following TKI discontinuation including immune surveillance and leukemic stem cell (LSC) depletion following TKI treatment. Data from relevant articles published on the Pubmed database between January 2007 and January 2020 have been included.: The results from the majority of TKI discontinuation studies show a consistent picture with approximately half of eligible patients achieving treatment free remission (TFR). However, reliable clinical predictors or biomarkers for the outcome of TKI discontinuation remain elusive and the mechanisms to explain the diversity of discontinuation success are not completely understood. Future studies will need to focus on attempts to increase the number of patients eligible for treatment discontinuation and will likely involve drug combinations including novel agents aimed at targeting the residual LSC population and enhancement of immune surveillance mechanisms.
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [2] |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [3] |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [4] |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [5] |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [6] |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [7] |
The RING finger E3 ubiquitin ligase Siah2 is implicated in control of diverse cellular biological events, including MAPK signaling and hypoxia. Here we demonstrate that Siah2 is subject to regulation by the deubiquitinating enzyme USP13. Overexpression of USP13 increases Siah2 stability by attenuating its autodegradation. Consequently, the ability of Siah2 to target its substrates prolyl hydroxylase 3 and Spry2 (Sprouty2) for ubiquitin-mediated proteasomal degradation is attenuated. Conversely, inhibition of USP13 expression with corresponding shRNA decreases the stability of both Siah2 and its substrate Spry2. Thus, USP13 limits Siah2 autodegradation and its ubiquitin ligase activity against its target substrates. Strikingly, the effect of USP13 on Siah2 is not mediated by its isopeptidase activity: mutations in its ubiquitin-binding sequences positioned within the ubiquitin-specific processing protease and ubiquitin-binding domains, but not within putative catalytic sites, abolish USP13 binding to and effect on Siah2 autodegradation and targeted ubiquitination. Notably, USP13 expression is attenuated in melanoma cells maintained under hypoxia, thereby relieving Siah2 inhibition and increasing its activity under low oxygen levels. Significantly, on melanoma tissue microarray, high nuclear expression of USP13 coincided with high nuclear expression of Siah2. Overall, this study identifies a new layer of Siah2 regulation mediated by USP13 binding to ubiquitinated Siah2 protein with a concomitant inhibitory effect on its activity under normoxia.
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [8] |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [9] |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [10] |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [11] |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [12] |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [13] |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [14] |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [15] |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [16] |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| [17] |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| {{custom_ref.label}} |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
/
| 〈 |
|
〉 |
