Quantitative proteomic and phosphoproteomic studies reveal novel 5-fluorouracil resistant targets in hepatocellular carcinoma

doi:10.1016/j.jprot.2019.103501

	Quantitative proteomic and phosphoproteomic studies reveal novel 5-fluorouracil resistant targets in hepatocellular carcinoma
	Liu, Zhen 1,2; Wang, Yan 1,2; Yao, Yating 1,2; Fang, Zheng 1,2; Miao, Qing R.3,4,5,6,7; Ye, Mingliang 1,2
刊名	JOURNAL OF PROTEOMICS
	2019-09-30
卷号	208 页码:10
关键词	Hepatocellular carcinoma 5-Fu resistance LC-MS/MS GnRH signaling pathway
ISSN号	1874-3919
DOI	10.1016/j.jprot.2019.103501
通讯作者	Miao, Qing R.(qing.miao@nyulangone.org) ; Ye, Mingliang(mingliang@dicp.ac.cn)
英文摘要	The development of chemoresistance remains the major obstacles to successful chemotherapy of hepatocellular carcinoma. The molecular mechanisms of drug resistance are complex. Identifying the key markers is crucial for development of therapeutic strategies to overcome resistance. In this study, we employed a cell-line model consisting of the 5-fluorouracil resistant Bel/5-Fu cell line and its parental Bel cell line. Using stable isotope dimethyl labeling combined with high-resolution mass spectrometry, in total, 8272 unique proteins and 22,095 phosphorylation sites with high localization confidence were identified. Our data indicated that the GnRH signaling pathway was involved in acquiring drug resistance, which has not been well elucidated. The western blotting results confirmed that the expression levels of PLC beta 3 and PLC beta 3 pS1105 in Bel/5-Fu cells were increased as compared to Bel cells. Furthermore, the protein levels of SRC and PKG delta, which could phosphorylate PLC beta 3 at ser1105, were higher in Bel/5-Fu cells than in Bel cells. The knockdown of SRC, PKC delta and PLC beta 3 increased the susceptibility of Bel/5-Fu cells to 5-Fu. Besides, the increased transcription levels of PLC beta 3, PRC delta and SRC were significantly associated with decreased overall survival. Together, our deep proteomic and phosphoproteomic data reveal novel therapeutic targets for attenuating 5-Fu resistance in anti-cancer therapy. Significance: It was reported that many hepatocellular carcinoma patients are resistance to 5-Fu. Although some studies related to drug resistance have been reported, the underlying mechanisms were not well elucidated. Unlike many single molecular studies, we focused on the global proteome and phosphoproteome analysis of Bel and Be15-/Fu cell line using stable isotope dimethyl labeling to identify the previously unrecognized signaling pathway for causing 5-Fu resistance. Our results showed that the phosphorylation levels of PLC beta 3 pS1105 and the protein levels of PLC beta 3, PKC delta and SRC, which are major components of GnRH signaling pathway were higher in Bel/5-Fu cells than in Bel cells. Furthermore, knockdown of PLC beta 3, PKC delta and SRC increased the susceptibility of Bel/5-Fu cells to 5-Fu. Overall, this is the first comprehensive proteomic and phosphoproteomic studies on 5-Fu resistant cell line Bel/5-Fu to identify the potential targets of attenuating chemoresistance in hepatocellular carcinoma.
资助项目	China State Key Basic Research Program[2016YFA0501402] ; China State Key Basic Research Program[2017YFA0505004] ; National Natural Science Foundation of China[21535008] ; National Natural Science Foundation of China[81600046] ; National Natural Science Foundation of China[81730016] ; National Natural Science Foundation of China[91753105] ; innovation program of science and research from the DICP, CAS[DICP TMSR201601] ; innovation program of science and research from the DICP, CAS[DICPQIBEBT UN201802] ; LiaoNing Revitalization Talents Program ; National Science Fund of China for Distinguished Young Scholars[21525524]
WOS关键词	FOCAL ADHESION KINASE ; PHOSPHORYLATION ; EXPRESSION ; NETWORKS ; PROTEINS ; THERAPY ; CELLS
WOS研究方向	Biochemistry & Molecular Biology
语种	英语
出版者	ELSEVIER
WOS记录号	WOS:000487172100017
资助机构	China State Key Basic Research Program ; China State Key Basic Research Program ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; innovation program of science and research from the DICP, CAS ; innovation program of science and research from the DICP, CAS ; LiaoNing Revitalization Talents Program ; LiaoNing Revitalization Talents Program ; National Science Fund of China for Distinguished Young Scholars ; National Science Fund of China for Distinguished Young Scholars ; China State Key Basic Research Program ; China State Key Basic Research Program ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; innovation program of science and research from the DICP, CAS ; innovation program of science and research from the DICP, CAS ; LiaoNing Revitalization Talents Program ; LiaoNing Revitalization Talents Program ; National Science Fund of China for Distinguished Young Scholars ; National Science Fund of China for Distinguished Young Scholars ; China State Key Basic Research Program ; China State Key Basic Research Program ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; innovation program of science and research from the DICP, CAS ; innovation program of science and research from the DICP, CAS ; LiaoNing Revitalization Talents Program ; LiaoNing Revitalization Talents Program ; National Science Fund of China for Distinguished Young Scholars ; National Science Fund of China for Distinguished Young Scholars ; China State Key Basic Research Program ; China State Key Basic Research Program ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; innovation program of science and research from the DICP, CAS ; innovation program of science and research from the DICP, CAS ; LiaoNing Revitalization Talents Program ; LiaoNing Revitalization Talents Program ; National Science Fund of China for Distinguished Young Scholars ; National Science Fund of China for Distinguished Young Scholars
内容类型	期刊论文
源URL	[http://cas-ir.dicp.ac.cn/handle/321008/172656]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
通讯作者	Miao, Qing R.; Ye, Mingliang
作者单位	1.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 2.Chinese Acad Sci, Natl Chromatog R&A Ctr, Dalian Inst Chem Phys, CAS Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China 3.Med Coll Wisconsin, Childrens Res Inst, Dept Surg, Div Pediat Surg, Milwaukee, WI 53226 USA 4.Med Coll Wisconsin, Childrens Res Inst, Dept Surg, Div Pediat Pathol, Milwaukee, WI 53226 USA 5.Med Coll Wisconsin, Childrens Res Inst, Dept Pathol, Div Pediat Surg, Milwaukee, WI 53226 USA 6.Med Coll Wisconsin, Childrens Res Inst, Dept Pathol, Div Pediat Pathol, Milwaukee, WI 53226 USA 7.NYU, Winthrop Hosp, Mineola, NY 11501 USA
推荐引用方式 GB/T 7714	Liu, Zhen,Wang, Yan,Yao, Yating,et al. Quantitative proteomic and phosphoproteomic studies reveal novel 5-fluorouracil resistant targets in hepatocellular carcinoma[J]. JOURNAL OF PROTEOMICS,2019,208:10.
APA	Liu, Zhen,Wang, Yan,Yao, Yating,Fang, Zheng,Miao, Qing R.,&Ye, Mingliang.(2019).Quantitative proteomic and phosphoproteomic studies reveal novel 5-fluorouracil resistant targets in hepatocellular carcinoma.JOURNAL OF PROTEOMICS,208,10.
MLA	Liu, Zhen,et al."Quantitative proteomic and phosphoproteomic studies reveal novel 5-fluorouracil resistant targets in hepatocellular carcinoma".JOURNAL OF PROTEOMICS 208(2019):10.