Email Alert | RSS

Journal of Peking University (Health Sciences) ›› 2020, Vol. 52 ›› Issue (2): 227-233. doi: 10.19723/j.issn.1671-167X.2020.02.006

Previous Articles     Next Articles

Palmitoylome profiling indicates that androgens promote the palmitoylation of metabolism-related proteins in prostate cancer-derived LNCaP cells

Wen-qing LI1,Si-mei REN2,3,△(),Xing-bo LONG3,4,Yu-qing TIAN1   

  1. 1. The Key Laboratory of Geriatrics, Beijing Institute of Geriatrics, Beijing Hospital, National Center of Gerontology, National Health Commission; Institute of Geriatric Medicine, Chinese Academy of Medical Science, Beijing 100730, China
    2. National Center for Clinical Laboratory, Beijing Hospital, National Center of Gerontology; Institute of Geriatric Medicine, Chinese Academy of Medical Science, Beijing 100730, China
    3. Graduate School of Chinese Academy of Medical Science and Peking Union Medical College, Beijing 100730, China
    4. Department of Urology, Beijing Hospital, Beijing 100730, China
  • Received:2019-12-10 Online:2020-04-18 Published:2020-04-18
  • Contact: Si-mei REN E-mail:rensimei4162@bjhmoh.cn
  • Supported by:
    Supported by the National Natural Science Foundation of China(81670161);Beijing Hospital Initial Foundation for Doctors(BJ2015-105)

RICH HTML

 24   

PDF (PC)

495

Abstract:

Objective: To explore potential therapeutic targets other than androgen-deprivation treatment for prostate cancer by screening the proteins induced by androgen at palmitoylation modification level in LNCaP cells.Methods: The LNCaP cells were treated with androgen (Methyltrienolone, R1881, 5 nmol/L) or dimethyl sulfoxide (DMSO) for 24 h, and then labeled with alkynyl palmitic acid Alk-C16 (100 μmol/L). After that, the cells were collected, lysed, the total protein was extracted, agarose beads labeled with azide (1 mmol/L) were added, and the click-chemistry reaction was carried out at room temperature for 1 h. The covalent bond formed by click-chemistry reaction of azide and alkynyl group was used to enrich the palmitoylated proteins on agarose beads. Label-free quantitation (LFQ) was used to compare the protein palmitoylation level of R1881 treated and untreated cells to screen the proteins induced by androgen at palmitoylation modification level.Results: In this experiment, 907 potential palmitoylated proteins (mascot score>2, P<0.05) were identified, among which 430 proteins had LFQ values not zero at least twice. Among the 430 proteins, the palmitoylation levels of 92 candidates were increased by androgen treatment, and their LFQ values were significantly upregulated (>1.5-fold, P<0.05) in ≥2 samples of androgen-treated vs. untreated LNCaP cells. We also used the software of cytoscape to classify the 92 proteins, and found that the known functional proteins of them could be divided into three categories: metabolism related, protein folding related and translation initiation related. Among them, metabolism related proteins included lipid metabolism (6), glucose metabolism (7) and respiratory electron transport chain (8), and a small amount of amino acid metabolism (2) and other metabolism related proteins (2). Notably, the ratio of LFQ of cytochrome b-c1 complex subunit 2 (UQCRC2) was significantly (>3-fold, P<0.05) higher in androgen-treated cells compared with untreated cells, indicating that the palmitoylation level of UQCRC2 was enhanced by androgen most significantly than that of others. The second was long-chain acyl CoA dehydrogenase (ACADVL) related to lipid metabolism and glucose 6-phosphate dehydrogenase (PGD) related to glucose metabolism, but the LFQ ratio of them was less than 3-fold.Conclusion: The research on palmitoylation mechanism of metabolism, especially the proteins related to respiratory electron transport chain, will provide a new guidance for the diagnosis and treatment of prostate cancer and the development of targeted drugs.

Key words: Prostate cancer, Androgen, Palmitoylation, Metabolism

CLC Number: 

  • R737.25

Figure 1

A schematic overview of screening of androgen-induced palmitoylated proteins"

Figure 2

Functional network of androgen-induced palmitoylated proteins in LNCaP cells Protein interaction network presenting the functions of androgen-induced palmitoylated proteins in metabolism, protein folding and translational initiation."

Figure 3

Androgen promoted the palmitoylation levels of metabolism-related proteins in LNCaP cells The numerical data of each group were presented as the mean ± standard deviation. The significant differences were analyzed using a paired student's t-test. P<0.05 was considered to indicate a statistically significant difference. *P<0.05; # P<0.01."

Figure 4

Androgen increased the palmitoylation level of UQCRC2 in LNCaP cellsPalm, palmitoylation; HAM, hydroxylamine; IP, immunoprecipitation; IB, immunoblotting; DMSO, dimethyl sulfoxide."

[1] Bray F, Ferlay J, Soerjomataram I , et al. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin, 2018,68(6):394-424.
[2] Chen W, Zheng R, Zhang S , et al. Cancer incidence and mortality in China, 2013[J]. Cancer Lett, 2017,401:63-71.
[3] Watson PA, Arora VK, Sawyers CL . Emerging mechanisms of resistance to androgen receptor inhibitors in prostate cancer[J]. Nat Rev Cancer, 2015,15(12):701-711.
[4] Wyatt AW, Gleave ME . Targeting the adaptive molecular landscape of castration-resistant prostate cancer[J]. EMBO Mol Med, 2015,7(7):878-894.
[5] Bishop JL, Davies A, Ketola K , et al. Regulation of tumor cell plasticity by the androgen receptor in prostate cancer[J]. Endocr Relat Cancer, 2015,22(3):R165-182.
[6] Quigley DA, Dang HX, Zhao SG , et al. Genomic hallmarks and structural variation in metastatic prostate cancer[J]. Cell, 2018,175(3):889.
[7] Viswanathan SR, Ha G, Hoff AM , et al. Structural alterations driving castration-resistant prostate cancer revealed by linked-read genome sequencing[J]. Cell, 2018,174(2):433-447.e419.
[8] Aicart-Ramos C, Valero RA, Rodriguez-Crespo I . Protein palmitoylation and subcellular trafficking[J]. Biochim Biophys Acta, 2011,1808(12):2981-2994.
[9] Charollais J , Van Der Goot FG. Palmitoylation of membrane proteins[J]. Mol Membr Biol, 2009,26(1):55-66.
[10] Ko PJ, Dixon SJ . Protein palmitoylation and cancer[J]. EMBO Rep, 2018,19(10):e46666.
[11] Thinon E, Hang HC . Chemical reporters for exploring protein acylation[J]. Biochem Soc Trans, 2015,43(2):253-261.
[12] Gottlieb CD, Linder ME . Structure and function of DHHC protein S-acyltransferases[J]. Biochem Soc Trans, 2017,45(4):923-928.
[13] Pepinsky RB, Zeng C, Wen D , et al. Identification of a palmitic acid-modified form of human sonic hedgehog[J]. J Biol Chem, 1998,273(22):14037-14045.
[14] Chen S, Zhu B, Yin C , et al. Palmitoylation-dependent activation of MC1R prevents melanomagenesis[J]. Nature, 2017,549(7672):399-403.
[15] Yao H, Lan J, Li C , et al. Inhibiting PD-L1 palmitoylation enhances T-cell immune responses against tumours[J]. Nat Biomed Eng, 2019,3(4):306-317.
[16] Hannoush RN, Sun J . The chemical toolbox for monitoring protein fatty acylation and prenylation[J]. Nat Chem Biol, 2010,6(7):498-506.
[17] Hernandez JL, Davda D, Majmudar JD , et al. Correlated S-palmitoylation profiling of snail-induced epithelial to mesenchymal transition[J]. Mol Biosyst, 2016,12(6):1799-1808.
[18] Hannoush RN, Arenas-Ramirez N . Imaging the lipidome: omega-alkynyl fatty acids for detection and cellular visualization of lipid-modified proteins[J]. ACS Chem Biol, 2009,4(7):581-587.
[19] Liu NQ, Braakman RB, Stingl C , et al. Proteomics pipeline for biomarker discovery of laser capture microdissected breast cancer tissue[J]. J Mammary Gland Biol Neoplasia, 2012,17(2):155-164.
[20] Li W, Li W, Zou L , et al. Membrane targeting of inhibitory Smads through palmitoylation controls TGF-beta/BMP signaling[J]. Proc Natl Acad Sci USA, 2017,114(50):13206-13211.
[21] Liberti MV, Locasale JW . The Warburg effect: how does it benefit cancer cells[J]. Trends Biochem Sci, 2016,41(3):287.
[22] Li C, Zhang G, Zhao L , et al. Metabolic reprogramming in cancer cells: glycolysis, glutaminolysis, and Bcl-2 proteins as novel therapeutic targets for cancer[J]. World J Surg Oncol, 2016,14(1):15.
[23] Warburg O . On the origin of cancer cells[J]. Science, 1956,123(3191):309-314.
[24] Koppenol W, Bounds P . The Warburg effect and metabolic efficiency: recrunching the numbers[J]. Science, 2009,324:1029-1033.
[25] Shen LF, Chen YJ, Liu KM , et al. Role of S-palmitoylation by ZDHHC13 in mitochondrial function and metabolism in liver[J]. Sci Rep, 2017,7(1):2182.
[26] Crofts AR, Hong S, Wilson C , et al. The mechanism of ubihydroquinone oxidation at the Qo-site of the cytochrome bc1 complex[J]. Biochim Biophys Acta, 2013,1827(11/12):1362-1377.
[27] 王福 . 基于细胞色素bc1复合物三维结构的新型Q0位点抑制剂的设计与合成[D]. 华中师范大学, 2012.
[1] BAI Gao-chen,SONG Yi,JIN Jie,YU Wei,HE Zhi-song. Clinical efficacy of docetaxel combined with carboplatin in patients with metastatic castration-resistant prostate cancer [J]. Journal of Peking University (Health Sciences), 2021, 53(4): 686-691.
[2] Kui-xia SUN,Cun-ling YAN,Zhi-yan LI,Ping LIU,Wei ZHANG,Qun HE. Clinical value of serum isoform [-2] proprostate-specific antigen and its derivatives in predicting aggressive prostate cancer [J]. Journal of Peking University (Health Sciences), 2020, 52(2): 234-239.
[3] Yong ZHANG,Chang LIU,Bin CHEN,Fan CHEN,Jin-yu DUAN,Meng-jun ZHANG,Jian JIAO. Associations of impaired glucose metabolism with chronic peridontitis in pre-diabetes patients [J]. Journal of Peking University(Health Sciences), 2020, 52(1): 71-76.
[4] Kuan-gen ZHANG,Yu-he ZHOU,Ya-kun SHAO,Fang MEI,Jiang-feng YOU,Bei-ying LIU,Fei PEI. Novel tumor metastasis suppressorgene LASS2/TMSG1 S248A mutant promotes invasion of prostate cancer cells through increasing ATP6V0C expression [J]. Journal of Peking University(Health Sciences), 2019, 51(2): 210-220.
[5] Yu-bing XIAO,Mu-yao GUO,Xiao-xia ZUO. Immunometabolism and systemic lupus erythematosus [J]. Journal of Peking University(Health Sciences), 2018, 50(6): 1120-1124.
[6] TANG Xu, ZHAO Wei-hong, SONG Qin-qin, YIN Hua-qi, DU Yi-qing, SHENG Zheng-zuo, WANG Qiang, ZHANG Xiao-wei, LI Qing, LIU Shi-jun, XU Tao. Influence of SOX10 on the proliferation and invasion of prostate cancer cells [J]. Journal of Peking University(Health Sciences), 2018, 50(4): 602-606.
[7] ZOU Peng-cheng, YANG Yi-feng, XU Xiao-yan LIU Bei-ying, MEI Fang, YOU Jiang-feng， LIU Qi-chen, PEI Fei . Silencing of vacuolar ATPase c subunit ATP6V0C inhibits invasion of prostate cancer cells [J]. Journal of Peking University(Health Sciences), 2017, 49(6): 937-947.
[8] HUA Ke-han, YANG Lei, ZHANG Xiao-wei, BAI Wen-jun, LI Qing, XU Tao. Complete androgen insensitivity syndrome associated with vesical fistula: a case report and literature review [J]. Journal of Peking University(Health Sciences), 2017, 49(4): 724-729.
[9] JI Guang-jie, HUANG Cong, SONG Gang, LI Xue-song, SONG Yi, ZHOU Li-qun. Predictive factor analysis of time to progression of castration-resistant prostate cancer after androgen deprivation therapy [J]. Journal of Peking University(Health Sciences), 2017, 49(4): 657-662.
[10] GONG Yan-qing, ZHANG Cui-jian, HE Shi-ming, LI Xue-song, ZhOU Li-qun, GUO Ying-lu. Nuclear export signal of androgen receptor regulated of androgen receptor stability in prostate cancer [J]. Journal of Peking University(Health Sciences), 2017, 49(4): 569-574.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
[1] . [J]. Journal of Peking University(Health Sciences), 2009, 41(4): 456 -458 .
[2] . [J]. Journal of Peking University(Health Sciences), 2009, 41(2): 125 -128 .
[3] . [J]. Journal of Peking University(Health Sciences), 2009, 41(2): 135 -140 .
[4] . [J]. Journal of Peking University(Health Sciences), 2009, 41(2): 158 -161 .
[5] . [J]. Journal of Peking University(Health Sciences), 2009, 41(2): 217 -220 .
[6] . [J]. Journal of Peking University(Health Sciences), 2009, 41(1): 52 -55 .
[7] . [J]. Journal of Peking University(Health Sciences), 2009, 41(1): 109 -111 .
[8] . [J]. Journal of Peking University(Health Sciences), 2009, 41(3): 297 -301 .
[9] . [J]. Journal of Peking University(Health Sciences), 2009, 41(5): 505 -515 .
[10] . [J]. Journal of Peking University(Health Sciences), 2009, 41(5): 599 -601 .
Copyright © Journal of Peking University (Health Sciences), All Rights Reserved.
Powered by Beijing Magtech Co. Ltd