浏览全部资源
扫码关注微信
环指蛋白14通过抑制HNF1α促进肝癌细胞增殖与侵袭转移
Ring finger protein 14 promotes proliferation, migration and invasion of hepatoma cells by inhibiting hepatocyte nuclear factor-1α
- 第三军医大学学报 2020年42卷第4期 页码：375-383
- 作者机构：
400042 重庆，陆军军医大学(第三军医大学)大坪医院消化内科
- 作者简介：
[陈东风，E-mail: chendf1981@126.com CHEN Dongfeng, E-mail: chendf1981@126.com
- 基金信息：国家自然科学基金青年科学基金(81802459)
DOI：10.16016/j.1000-5404.201910077
中图分类号： R341; R730.23; R735.7收稿：2019-10-14，
修回：2019-11-02，
纸质出版：2020-02-28
- 稿件说明：
移动端阅览
朱光曦, 王文生, 张亮, 等. 环指蛋白14通过抑制HNF1α促进肝癌细胞增殖与侵袭转移[J]. 第三军医大学学报, 2020,42(4):375-383.
Guangxi ZHU, Wensheng WANG, Liang ZHANG, et al. Ring finger protein 14 promotes proliferation, migration and invasion of hepatoma cells by inhibiting hepatocyte nuclear factor-1α[J]. Journal of Third Military Medical University, 2020, 42(4): 375-383.
朱光曦, 王文生, 张亮, 等. 环指蛋白14通过抑制HNF1α促进肝癌细胞增殖与侵袭转移[J]. 第三军医大学学报, 2020,42(4):375-383. DOI： 10.16016/j.1000-5404.201910077.
Guangxi ZHU, Wensheng WANG, Liang ZHANG, et al. Ring finger protein 14 promotes proliferation, migration and invasion of hepatoma cells by inhibiting hepatocyte nuclear factor-1α[J]. Journal of Third Military Medical University, 2020, 42(4): 375-383. DOI： 10.16016/j.1000-5404.201910077.
摘要
目的
2
研究环指蛋白14(ring finger protein 14，RNF14)对肝癌细胞增殖与侵袭转移能力的影响，并探讨RNF14在肝癌发生、发展中的作用机制。
方法
2
[利用基因表达谱交互式分析网站(Gene Expression Profiling Interactive Analysis
GEPIA；
http://gepia.cancer-pku.cn
http://gepia.cancer-pku.cn
)与Western blot实验分析正常人肝细胞与肝癌细胞间RNF14的表达差异；应用Transwell小室迁移与侵袭实验研究过表达和敲低RNF14对肝癌细胞(Hu7、7404)运动迁移能力的影响；通过CCK-8增殖实验、克隆形成实验及裸鼠皮下成瘤实验体外、体内共同验证RNF14对肝癌细胞7404增殖能力的影响；通过免疫共沉淀实验检测RNF14与肝细胞核因子1α(hepatocyte nuclear factor-1alpha，HNF1α)之间的相互作用。
结果
2
GEPIA与Western blot实验结果显示:RNF14在肝癌组织和肝癌细胞中的表达分别显著高于癌旁组织和正常肝细胞(
P
<
0.05)。Transwell侵袭迁移实验结果显示:过表达RNF14后，肝癌细胞Hu7的迁移与侵袭能力显著高于阴性对照组(
P
<
0.05)，同样地，敲低RNF14后，肝癌细胞7404的迁移与侵袭能力显著低于阴性对照组(
P
<
0.05)。CCK-8增殖实验、克隆形成与皮下移植瘤实验结果证明:敲低RNF14能够显著抑制肝癌细胞的增殖(
P
<
0.05)。免疫共沉淀实验结果显示:RNF14与HNF1α之间存在相互结合，且RNF14能够下调HNF1α的表达。
结论
2
RNF14能够通过抑制抑癌因子HNF1α的表达，增强肝癌细胞的增殖与侵袭转移能力，从而在肝癌的发生、发展中起到重要作用。
Abstract
Objective
2
To investigate the effects of ring finger protein 14 (RNF14) overexpression and knockdown on the proliferation and invasion of hepatoma cells and explore the mechanism by which RNF14 promote the occurrence and progression of hepatocellular carcinoma (HCC).
Methods
2
We investigated the expression of RNF14 in HCC and adjacent tissues using Gene Expression Profiling Interactive Analysis (GEPIA)
and examined its expression in normal human hepatocytes and hepatoma cells using Western blotting. The effects of overexpression and knockdown of RNF14 on the migration and invasion abilities of 2 hepatoma cell lines (Hu7 and 7404) were observed using Transwell chamber migration and invasion assay. The effect of RNF14 knockdown on the proliferation of hepatoma cells 7404 was verified by CCK8 proliferation assay
colony formation assay and subcutaneous xenograft tumor formation in nude mice. The interaction between RNF14 and hepatocyte nuclear factor-1α (HNF1α) was tested with co-immunoprecipitation assay.
Results
2
The results of GEPIA and Western blotting showed that the expression of RNF14 was significantly higher in hepatoma tissues and cells than in normal hepatic tissues and hepatocytes (
P
<
0.05). Transwell chamber invasion and migration assays showed that the migration and invasion abilities of Hu7 cells were increased significantly after overexpression of RNF14 (
P
<
0.05)
and RNF14 knockdown significantly attenuated the migration and invasion ability of hepatoma cells 7404 (
P
<
0.05). The results of CCK8 proliferation assay
colony formation and subcutaneous xenograft tumor formation in nude mice all demonstrated that RNF14 knockdown significantly inhibited the proliferation of hepatoma cells (
P
<
0.05). Co-immunoprecipitation confirmed that RNF14 could bind to HNF1α and down-regulate the expression of the latter.
Conclusion
2
RNF14 promotes the proliferation
migration and invasion of hepatoma cells through degrading HNF1α
and may thus play an important role in the occurrence and progression of HCC.
关键词
Keywords
references
LAFARO K J, DEMIRJIAN A N, PAWLIK T M. Epidemi-ology of hepatocellular carcinoma[J]. Surg Oncol Clin N Am, 2015, 24(1): 1-17. DOI: 10.1016/j.soc.2014.09.001.
中华人民共和国卫生和计划生育委员会医政医管局.原发性肝癌诊疗规范(2017年版)[J].中华消化外科杂志, 2017, 16(7): 635-647. DOI: 10.3760/cma.j.issn.1673-9752.2017.07.001.
Bureau of Medical Administration.National Health and Family Planning Commission of the PRC. Standardization of diagnosis and treatment for hepatocellular carcinoma (2017 edition)[J]. Chin J Diges Surg, 2017. 16(7): 635-647. DOI: 10.3760/cma.j.issn.1673-9752.2017.07.001.
MCGLYNN K A, PETRICK J L, LONDON W T. Global epidemiology of hepatocellular carcinoma: an emphasis on demographic and regionalvariability[J]. Clin Liver Dis, 2015, 19(2): 223-238. DOI: 10.1016/j.cld.2015.01.001.
MALUCCIO M, COVEY A. Recent progress in understanding, diagnosing, and treating hepatocellularcarcinoma[J]. CA Cancer J Clin, 2012, 62(6): 394-399. DOI: 10.3322/caac.21161.
FOMER A, LLOVET J M, BRUIX J. Hepatocellular carcinoma[J]. Lancet, 2012, 379(9822): 1245-1255. DOI: 10.1016/S0140-6736(11)61347-0.
KANG H Y, YEH S, FUJIMOTO N, et al . Cloning and characterization of human prostate coactivator ARA54, a novel protein that associates with the androgen receptor[J ] . J Biol Chem, 1999, 274(13): 8570-8576. DOI: 10.1074/jbc.274.13.8570.
INGHAM A B, OSBORNE S A, MENZIES M, et al . RNF14 is a regulator of mitochondrial and immune function in muscle[J ] . BMC Syst Biol, 2014, 8: 10. DOI: 10.1186/1752-0509-8-10.
LIU X, YU X P, ZACK D J, et al . TiGER: a database for tissue-specific gene expression and regulation[J ] . BMC Bioinformatics, 2008, 9: 271. DOI: 10.1186/1471-2105-9-271.
WANG X W, WEI W, WANG W Q, et al . RING finger proteins are involved in the progression of barrett esophagus to esophageal adenocarcinoma: a preliminary study[J ] . Gut Liver, 2014, 8(5): 487-494. DOI: 10.5009/gnl13133.
WU B B, PILOTO S, ZENG W H, et al . Ring Finger Protein 14 is a new regulator of TCF/β-catenin-mediated transcription and colon cancer cell survival[J ] . EMBO Rep, 2013, 14(4): 347-355. DOI: 10.1038/embor.2013.19.
ODOM D T, ZIZLSPERGER N, GORDON D B, et al . Control of pancreas and liver gene expression by HNF transcription factors[J ] . Science, 2004, 303(5662): 1378-1381. DOI: 10.1126/science.1089769.
COSTA R H, KALINICHENKO VV, HOLTERMAN A X, et al . Transcription factors in liver development, differentiation, and regeneration[J ] . Hepatology, 2003, 38(6): 1331-1347. DOI: 10.1016/j.hep.2003.09.034.
ZENG X, LIN Y, YIN C, et al . Recombinant adenovirus carrying the hepatocyte nuclear factor-1alpha gene inhibits hepatocellular carcinoma xenograft growth in mice[J ] . Hepatology, 2011, 54(6): 2036-2047. DOI: 10.1002/hep.24647.
ZHU S J, HOSHIDA Y. Molecular heterogeneity in hepatocellular carcinoma[J]. Hepat Oncol, 2018, 5(1): HEP10. DOI: 10.2217/hep-2018-0005.
UEKI N, SEKI N, YANO K, et al . Isolation and characterization of a novel human gene (HFB30) which encodes a protein with a RING finger motif[J ] . Biochim Biophys Acta, 1999, 1445(2): 232-236. DOI: 10.1016/s0167-4781(99)00045-7.
LUCAS J I, ARNAU V, MARÍN I. Comparative genomics and protein domain graph analyses link ubiquitination and RNA metabolism[J]. J Mol Biol, 2006, 357(1): 9-17. DOI: 10.1016/j.jmb.2005.12.068.
YANG Z M, CHANG Y J, MIYAMOTO H, et al . Transgelin functions as a suppressor via inhibition of ARA54-enhanced androgen receptor transactivation and prostate cancer cell growth[J ] . Mol Endocrinol, 2007, 21(2): 343-358. DOI: 10.1210/me.2006-0104..
MIYAMOTO H, RAHMAN M, TAKATERA H, et al . A dominant-negative mutant of androgen receptor coregulator ARA54 inhibits androgen receptor-mediated prostate cancer growth[J ] . J Biol Chem, 2002, 277(7): 4609-4617. DOI: 10.1074/jbc.M108312200.
WINTER S L, BOSNOYAN-COLLINS L, PINNADUWAGE D, et al . The interaction of PP1 with BRCA1 and analysis of their expression in breast tumors[J ] . BMC Cancer, 2007, 7: 85. DOI: 10.1186/1471-2407-7-85.
KIKUCHI H, UCHIDA C, HATTORI T, et al . ARA54 is involved in transcriptional regulation of the cyclin D1 gene in human cancer cells[J ] . Carcinogenesis, 2007, 28(8): 1752-1758. DOI: 10.1093/carcin/bgm120.
KALASEKAR S M, KOTIYAL S, CONLEY C, et al . Heterogeneous beta-catenin activation is sufficient to cause hepatocellular carcinoma in zebrafish[J ] . Biol Open, 2019, 8(10): bio047829. DOI: 10.1242/bio.047829.
ITO K, ADACHI S, IWAKAMI R, et al . N-Terminally extended human ubiquitin-conjugating enzymes (E2s) mediate the ubiquitination of RING-finger proteins, ARA54 and RNF8[J ] . Eur J Biochem, 2001, 268(9): 2725-2732. DOI: 10.1046/j.1432-1327.2001.02169.x.
DING F B, XIAO H B, WANG M S, et al . The role of the ubiquitin-proteasome pathway in cancer development and treatment[J ] . Front Biosci (Landmark Ed), 2014, 19: 886-895. DOI: 10.2741/4254.
SUN Y. E3 ubiquitin ligases as cancer targets and biomarkers[J]. Neoplasia, 2006, 8(8): 645-654. DOI: 10.1593/neo.06376.
LONG D X, WANG P, SUN Y J, et al . Neuropathy target esterase is degraded by the ubiquitin-proteasome pathway with ARA54 as the ubiquitin ligase[J ] . Biochemistry, 2015, 54(50): 7385-7392. DOI: 10.1021/acs.biochem.5b00879.
NI Q, DING K, WANG K Q, et al . Deletion of HNF1α in hepatocytes results in fatty liver-related hepatocellular carcinoma in mice[J ] . FEBS Lett, 2017, 591(13): 1947-1957. DOI: 10.1002/1873-3468.12689.
The UniProt Consortium. UniProt: the universal protein knowledgebase[J]. Nucleic Acids Res, 2017, 45(D1): D158-D169. DOI: 10.1093/nar/gkw1099.
0
浏览量
0
下载量
0
CSCD
- 网络PDF下载
- Meta-XML下载
- BibTeX下载
- Endnote下载
- RefMan 下载
- NoteExpress下载
- NoteFirst下载
关联资源
相关文章
相关作者
相关机构
- 技术支持由北京北大方正电子有限公司提供 京ICP备09064830号-19
京公网安备11010802024621 - 本系统建议在Chrome、 IE9+ 以上版本浏览器阅读本站内容，360浏览器请切换至极速模式
- Cookies帮助我们提供服务并提供个性化体验。使用本网站，即表示您同意我们使用Cookies
