Knockdown of Deubiquitinase USP13 Inhibits the Proliferation of K562 Cells

doi:10.13523/j.cb.2101041

China Biotechnology

2021, Vol. 41

Issue (5): 1-7 DOI: 10.13523/j.cb.2101041

Knockdown of Deubiquitinase USP13 Inhibits the Proliferation of K562 Cells

TAO Shou-song¹,REN Guang-ming²,YIN Rong-hua²,YANG Xiao-ming^1,²,MA Wen-bing^3,^**(

),GE Zhi-qiang^1,^**(

)

1 Department of Pharmaceutical Engineering, Tianjin University, Tianjin 300072, China
2 Beijing Institute of Lifeomics, Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 100850, China
3 Institute of Health Service and Blood Academy of Military Medical Sciences, Beijing 100850, China

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Abstract

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.

Key words： Chronic myelogenous leukemia K562 cells USP13 Proliferation and apoptosis c-Myc

Received: 27 January 2021 Published: 01 June 2021

ZTFLH:

Q814

Corresponding Authors: Wen-bing MA,Zhi-qiang GE E-mail: mawenbing003@163.com;gezhiq@tju.edu.cn

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	Shou-song TAO
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	Rong-hua YIN
	Xiao-ming YANG
	Wen-bing MA
	Zhi-qiang GE

Cite this article:

TAO Shou-song,REN Guang-ming,YIN Rong-hua,YANG Xiao-ming,MA Wen-bing,GE Zhi-qiang. Knockdown of Deubiquitinase USP13 Inhibits the Proliferation of K562 Cells. China Biotechnology, 2021, 41(5): 1-7.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.2101041 OR https://manu60.magtech.com.cn/biotech/Y2021/V41/I5/1

Table 1 Primer name and sequence

Fig.1 High expression of USP13 in K562 cells Q-PCR (a) and Western blot (b) analysis the expression levels of USP13 in K562, MCF-7 and SW872 cells

Fig.2 Construction of pLKO.1-shUSP13-GFP vector and lentivirus packaged (a) Sequencing results of pLKO.1-shUSP13-GFP vector (b) Western blot analysis knockdown efficiency of USP13 in HEK293T cells (c) Fluorescence microscopy detects the lentivirus packaged in HEK293T cells

Fig.3 Construction of stable USP13 knockdown K562 cell lines (a) Flow cytometry analysis of GFP rate in K562 cells (b) Q-PCR and (c) Western blot analysis the knockdown efficiency of USP13 in K562 cells

Fig.4 Knockdown of USP13 promotes K562 cell apoptosis and inhibits cell proliferation Flow cytometry analysis of cell proliferation (a) or apoptosis (b)

Fig.5 Knockdown of USP13 decreases c-Myc level by enhancing its ubiquitinations (a) Western blot analysis of c-Myc levels in USP13 knockdown K562 cells (b) Co-immunoprecipitation analysis the interaction between USP13 and c-Myc in K562 cells (c) Detection the ubiquitination of c-Myc in USP13 knockdown K562 cells (d) c-Myc is degraded through the ubiquitin-proteasome pathway


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