[1]张 驰,卢 山.USP13上调STAT1抑制前列腺癌细胞增殖、克隆和迁移[J].南京师范大学学报(工程技术版),2021,21(04):072-79.[doi:10.3969/j.issn.1672-1292.2021.04.011]
 Zhang Chi,Lu Shan.USP13 Inhibits Proliferation,Cloning,and Migration of ProstateCancer Cells by Upregulating STAT1[J].Journal of Nanjing Normal University(Engineering and Technology),2021,21(04):072-79.[doi:10.3969/j.issn.1672-1292.2021.04.011]
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USP13上调STAT1抑制前列腺癌细胞增殖、克隆和迁移
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南京师范大学学报(工程技术版)[ISSN:1006-6977/CN:61-1281/TN]

卷:
21卷
期数:
2021年04期
页码:
072-79
栏目:
生物工程
出版日期:
2021-12-15

文章信息/Info

Title:
USP13 Inhibits Proliferation,Cloning,and Migration of ProstateCancer Cells by Upregulating STAT1
文章编号:
1672-1292(2021)04-0072-08
作者:
张 驰12卢 山12
(1.南京师范大学生命科学学院,江苏 南京 210023)(2.南京师范大学江苏省分子医学生物技术重点实验室,江苏 南京 210023)
Author(s):
Zhang Chi12Lu Shan12
(1.School of Life Sciences,Nanjing Normal University,Nanjing 210023,China)(2.Jiangsu Key Laboratory for Molecular and Medical Biotechnology,Nanjing Normal University,Nanjing 210023,China)
关键词:
前列腺癌USP13去泛素化迁移STAT1
Keywords:
prostate cancerUSP13deubiquitinationmigrationSTAT1
分类号:
R737.25
DOI:
10.3969/j.issn.1672-1292.2021.04.011
文献标志码:
A
摘要:
为了探究去泛素化酶USP13在前列腺癌中发挥的功能及其相关分子机制,采用PCR、酶切、酶连和转化大肠杆菌等方法,成功构建USP13基因真核表达重组质粒. 同时,在前列腺癌细胞PC-3和DU145中转染USP13质粒,进行CCK-8实验、克隆形成、划痕以及Transwell迁移实验; 利用逆转录PCR、蛋白质免疫印迹法和泛素化Co-IP等实验,分析USP13过表达对信号转导与转录激活因子 1(signal transducer and activator of transcription 1,STAT1)的影响. 实验结果发现,USP13可以抑制前列腺癌细胞的增殖、克隆和迁移能力; 过表达USP13不影响STAT1 mRNA水平,但上调STAT1蛋白水平,而敲低USP13则下调STAT1蛋白水平; 同时,过表达USP13降低了STAT1泛素化水平,敲低USP13则提高STAT1泛素化水平.
Abstract:
In order to explore the role of deubiquitination enzyme USP13 in prostate cancer cells and its regulatory pathway,the eukaryotic expression plasmid of USP13 is successfully constructed via PCR,enzyme digestion,enzyme ligation and transformation into Escherichia coli. Meanwhile,CCK-8 assay,colony formation assay,wound healing assay and Transwell migration assay are performed by transfecting USP13 plasmid into PC-3 and DU145 cells. Then the effects of USP13 overexpression on mRNA experssion,protein expression and ubiquition level of signal transducer and activator of transcription 1(STAT1)are detected by reverse transcription PCR,western blotting and ubiquitination Co-IP assay,respectively. USP13 is found to inhibit the growth,cloning and migration of prostate cancer cells,which has an effect on STAT1 protein level but not on transcription level of STAT1. Overexpression of USP13 upregulates STAT1 protein level,while knockdown of USP13 decreases STAT1 protein level. Furthermore,overexpression of USP13 decreases the level of STAT1 ubiquitination,while knockdown of USP13 increases the level of STAT1 ubiquitination.

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备注/Memo

备注/Memo:
收稿日期:2021-06-09.
基金项目:国家自然科学基金项目(81272850、81472415).
通讯作者:卢山,博士,教授,研究方向:前列腺肿瘤侵袭转移的分子机制及调控. E-mail:lu_shan@hotmail.com
更新日期/Last Update: 2021-12-15