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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2022, Vol. 42 Issue (5): 27-36    DOI: 10.13523/j.cb.2112042
研究报告     
XIAP介导PTEN neddylation修饰促进结肠癌细胞增殖和迁移*
袁淑辉1,3,李少华2,方威2,彭志强3,张令强1,3,**()
1 青岛大学基础医学院 青岛 266071
2 南方医科大学第三临床医学院上海市奉贤区中心医院 上海 201499
3 军事科学院军事医学研究院生命组学研究所 蛋白质组学国家重点实验室 北京 100850
XIAP Mediated-PTEN Neddylation Promotes Proliferation and Migration of Colon Cancer Cells
YUAN Shu-hui1,3,LI Shao-hua2,FANG Wei2,PENG Zhi-qiang3,ZHANG Ling-qiang1,3,**()
1 School of Basic Medicine, Qingdao 266071, China
2 Shanghai Fengxian Central Hospital, The Third School of Clinical Medicine, Southern Medical University, Shanghai 201499, China
3 State Key Laboratory of Proteomics, Beijing National Center for Protein Sciences (Beijing), Beijing Institute of Lifeomics, Beijing 100850, China
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摘要：

目的:探讨XIAP(X-linked Apoptosis Inhibitor of Protein)介导的PTEN neddylation修饰对结肠癌细胞系SW480增殖和迁移能力的影响以及临床意义。方法:利用免疫组织化学技术对人结直肠癌及癌旁组织芯片中neddylation E3连接酶XIAP的表达情况进行检测,通过Western blot验证临床标本中人结直肠癌与癌旁组织中XIAP的表达情况;利用Co-IP检测SW480细胞系内源PTEN与XIAP的相互作用;利用CRISPR Cas9技术构建XIAP敲除的SW480细胞系,免疫沉淀技术检测XIAP 敲除的SW480细胞系中PTEN的neddylation修饰水平;变性条件下裂解临床结直肠癌与癌旁组织,利用免疫沉淀技术检测人结直肠癌与癌旁组织中PTEN 的 neddylation修饰水平;分别在XIAP敲除的SW480 细胞系与对照组 SW480 细胞系中回转或不回转FLAG-PTEN-Nedd8融合质粒来模拟恢复或者不恢复 SW480 细胞系中 PTEN的neddylation修饰水平,分别采用CCK8与Transwell实验探索XIAP-PTEN neddylation调控轴对SW480 细胞增殖与迁移的影响。结果:结直肠癌组织中XIAP表达高于癌旁组织;结肠癌细胞SW480中PTEN与XIAP具有相互作用;敲除XIAP抑制了PTEN neddylation修饰水平;结直肠癌组织PTEN neddylation修饰水平高于癌旁组织;敲除XIAP显著抑制SW480细胞的增殖和迁移能力,而过表达FLAG-PTEN-Nedd8融合质粒可挽救这种抑制作用。结论:在结直肠癌中,XIAP的蛋白质表达水平明显高于癌旁组织;XIAP介导的PTEN neddylation修饰水平明显高于癌旁组织。同时XIAP主要通过介导PTEN neddylation修饰促进了结肠癌细胞SW480的增殖和迁移。
关键词： 结直肠癌;  XIAP;  PTEN neddylation;  增殖;  迁移    
Abstract:

Objective: To explore the functional role of XIAP-PTEN neddylation axis in colorectal cancer at cell line and clinical level. Methods: The expression level of XIAP was analyzed in tissue microarray of colorectal cancer by immunohistochemical staining, and the protein level of XIAP was detected in human colorectal cancer and adjacent normal tissues by Western blot. Co-IP was performed to analyze the interaction between endogenous PTEN and XIAP in SW480 cell line. CRISPR Cas9 technology was used to construct XIAP-knockout SW480 cell line, and the neddylation level of PTEN was analyzed in sg-XIAP cells by immunoprecipitation and Western blot. XIAP knockout or wild type SW480 cell lines were co-transfected with FLAG-Vector or FLAG-PTEN-Nedd8 plasmid, and then CCK8 and Transwell experiments were used to assay the proliferation and migration of XIAP-PTEN neddylation axis in SW480 cells, respectively. Results: The expression levels of XIAP were up-regulated in colon and rectum cancer tissues compared with adjacent normal tissues. XIAP interacted with PTEN in SW480 colon cancer cells. Deletion of XIAP inhibited PTEN neddylation in SW480 cells. The level of PTEN neddylation was elevated in colorectal cancer tissues compared with adjacent normal tissues. Deletion of XIAP inhibited the proliferation and migration of SW480 cells significantly, while PTEN-Nedd8 fusion protein rescued the phenotypes of XIAP deletion in SW480 cells. Conclusion: XIAP-PTEN neddylation axis promotes SW480 colon cancer cell proliferation and migration.
Key words: Colorectal cancer    XIAP    PTEN neddylation    Proliferation    Migration
收稿日期: 2021-12-19 出版日期: 2022-06-17
ZTFLH:  Q28  
基金资助: *国家自然科学基金面上项目(81974428)
通讯作者: 张令强     E-mail: zhanglq@nic.bmi.ac.cn
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引用本文:

袁淑辉,李少华,方威,彭志强,张令强. XIAP介导PTEN neddylation修饰促进结肠癌细胞增殖和迁移*[J]. 中国生物工程杂志, 2022, 42(5): 27-36.

YUAN Shu-hui,LI Shao-hua,FANG Wei,PENG Zhi-qiang,ZHANG Ling-qiang. XIAP Mediated-PTEN Neddylation Promotes Proliferation and Migration of Colon Cancer Cells. China Biotechnology, 2022, 42(5): 27-36.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.2112042    或    https://manu60.magtech.com.cn/biotech/CN/Y2022/V42/I5/27

图1  XIAP在结直肠癌中高表达
图2  结直肠癌中XIAP表达与PTEN neddylation水平显著正相关
图3  XIAP通过介导PTEN neddylation修饰促进结直肠癌细胞的增殖和迁移
图4  MLN4924和C75联合用药协同抑制SW480细胞的增殖及迁移
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