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Current Molecular Medicine

Editor-in-Chief

ISSN (Print): 1566-5240
ISSN (Online): 1875-5666

Research Article

Ubiquitin Mediated Degradation of EGFR by 17 β-estradiol in Triple Negative MDA-MB-231 (TNBC) Breast Cancer Cells Line

Author(s): Vitthal Khode*, Sumangala Patil, Vishwas Kaveeshwar, Komal Ruikar, Anil Bargale, Sarathkumar E. and Satish Patil

Volume 22, Issue 5, 2022

Published on: 29 July, 2021

Page: [449 - 457] Pages: 9

DOI: 10.2174/1566524021666210729144713

Abstract

Background: Triple Negative Breast Cancer (TNBC) commonly displays Epidermal growth factor receptor (EGFR). Effective EGFR degradation results in the suppression of tumor in various models. Studies have addressed the relevance of this strategy in the treatment of TNBC. In the present study, we examined the effect of 17 β- estradiol on EGFR expression in MDA-MB-231 (TNBC) cell line and assessed whether 17 β-estradiol degrades EGFR by ubiquitination pathway.

Objectives: The objective of this study is to treat MDA-MB-231 cell lines with Cycloheximide with or without 17β-estrdiol to observe whether 17β-estradiol leads to EGFR degradation and to treat with MG-132 to assess whether degradation occurs through ubiquitination pathway.

Methods: MDA-MB-231 cells were treated with 17β-estradiol (E2) and EGFR expression was studied by western blotting at different intervals by using Cycloheximide chase. To assess ubiquitination pathway of degradation of EGFR in MDA-MB-231 cell line, MG-132 was used.

Results: EGFR expression was reduced with β-estradiol treatment in MDA-MB-231 cell line with Cycloheximide chase. Upon Treatment with MG-132 and E2, EGFR expression did not reduce, suggesting that Estrogen degrades EGFR by ubiquitination pathway.

Conclusion: Estrogen degrades EGFR in MDA-MB-231 cells and this degradation occurs by ubiquitination.

Keywords: Triple negative breast cancer, MDA-MB-231, estrogen, epidermal growth factor receptor, MG-132, ubiquitination.

[1]
Foulkes WD, Smith IE, Reis-Filho JS. Triple-negative breast cancer. N Engl J Med 2010; 363(20): 1938-48.
[http://dx.doi.org/10.1056/NEJMra1001389] [PMID: 21067385]
[2]
Morris GJ, Naidu S, Topham AK, et al. Differences in breast carcinoma characteristics in newly diagnosed African-American and Caucasian patients: a single-institution compilation compared with the National Cancer Institute’s Surveillance, Epidemiology, and End Results database. Cancer 2007; 110(4): 876-84.
[http://dx.doi.org/10.1002/cncr.22836] [PMID: 17620276]
[3]
Bolla M, Chedin M, Souvignet C, Marron J, Arnould C, Chambaz E. Estimation of epidermal growth factor receptor in 177 breast cancers: correlation with prognostic factors. Breast Cancer Res Treat 1990; 16(2): 97-102.
[http://dx.doi.org/10.1007/BF01809293] [PMID: 2176111]
[4]
Slamon DJ, Godolphin W, Jones LA, et al. Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer. Science 1989; 244(4905): 707-12.
[http://dx.doi.org/10.1126/science.2470152] [PMID: 2470152]
[5]
Rimawi MF, Shetty PB, Weiss HL, et al. Epidermal growth factor receptor expression in breast cancer association with biologic phenotype and clinical outcomes. Cancer 2010; 116(5): 1234-42.
[http://dx.doi.org/10.1002/cncr.24816] [PMID: 20082448]
[6]
Nicholson RI, Hutcheson IR, Jones HE, et al. Growth factor signalling in endocrine and anti-growth factor resistant breast cancer. Rev Endocr Metab Disord 2007; 8(3): 241-53.
[http://dx.doi.org/10.1007/s11154-007-9033-5] [PMID: 17486454]
[7]
DeSantis CE, Fedewa SA, Goding Sauer A, Kramer JL, Smith RA, Jemal A. Breast cancer statistics, 2015: Convergence of incidence rates between black and white women. CA Cancer J Clin 2016; 66(1): 31-42.
[http://dx.doi.org/10.3322/caac.21320] [PMID: 26513636]
[8]
Brewster AM, Chavez-MacGregor M, Brown P. Epidemiology, biology, and treatment of triple-negative breast cancer in women of African ancestry. Lancet Oncol 2014; 15(13): e625-34.
[http://dx.doi.org/10.1016/S1470-2045(14)70364-X] [PMID: 25456381]
[9]
Spangler JB, Neil JR, Abramovitch S, et al. Combination antibody treatment down-regulates epidermal growth factor receptor by inhibiting endosomal recycling. Proc Natl Acad Sci USA 2010; 107(30): 13252-7.
[http://dx.doi.org/10.1073/pnas.0913476107] [PMID: 20616078]
[10]
Goh LK, Huang F, Kim W, Gygi S, Sorkin A. Multiple mechanisms collectively regulate clathrin-mediated endocytosis of the epidermal growth factor receptor. J Cell Biol 2010; 189(5): 871-83.
[http://dx.doi.org/10.1083/jcb.201001008] [PMID: 20513767]
[11]
Crowder JJ, Geigges M, Gibson RT, et al. Rkr1/Ltn1 Ubiquitin Ligase-mediated Degradation of Translationally Stalled Endoplasmic Reticulum Proteins. J Biol Chem 2015; 290(30): 18454-66.
[http://dx.doi.org/10.1074/jbc.M115.663559] [PMID: 26055716]
[12]
Tran JR, Brodsky JL. Assays to measure ER-associated degradation in yeast. Methods Mol Biol 2012; 832: 505-18.
[http://dx.doi.org/10.1007/978-1-61779-474-2_36] [PMID: 22350909]
[13]
Kao SH, Wang WL, Chen CY, et al. GSK3β controls epithelial-mesenchymal transition and tumor metastasis by CHIP-mediated degradation of Slug. Oncogene 2014; 33(24): 3172-82.
[http://dx.doi.org/10.1038/onc.2013.279] [PMID: 23851495]
[14]
Hampton RY, Rine J. Regulated degradation of HMG-CoA reductase, an integral membrane protein of the endoplasmic reticulum, in yeast. J Cell Biol 1994; 125(2): 299-312.
[http://dx.doi.org/10.1083/jcb.125.2.299] [PMID: 8163547]
[15]
Hochstrasser M, Varshavsky A. In vivo degradation of a transcriptional regulator: the yeast alpha 2 repressor. Cell 1990; 61(4): 697-708.
[http://dx.doi.org/10.1016/0092-8674(90)90481-S] [PMID: 2111732]
[16]
Hanna J, Leggett DS, Finley D. Ubiquitin depletion as a key mediator of toxicity by translational inhibitors. Mol Cell Biol 2003; 23(24): 9251-61.
[http://dx.doi.org/10.1128/MCB.23.24.9251-9261.2003] [PMID: 14645527]
[17]
Glickman MH, Ciechanover A. The ubiquitin-proteasome proteolytic pathway: destruction for the sake of construction. Physiol Rev 2002; 82(2): 373-428.
[http://dx.doi.org/10.1152/physrev.00027.2001] [PMID: 11917093]
[18]
Hershko A. The ubiquitin system for protein degradation and some of its roles in the control of the cell division cycle. Cell Death Differ 2005; 12(9): 1191-7.
[http://dx.doi.org/10.1038/sj.cdd.4401702] [PMID: 16094395]
[19]
Ravid T, Hochstrasser M. Diversity of degradation signals in the ubiquitin-proteasome system. Nat Rev Mol Cell Biol 2008; 9(9): 679-90.
[http://dx.doi.org/10.1038/nrm2468] [PMID: 18698327]
[20]
Gallastegui N, Groll M. The 26S proteasome: assembly and function of a destructive machine. Trends Biochem Sci 2010; 35(11): 634-42.
[http://dx.doi.org/10.1016/j.tibs.2010.05.005] [PMID: 20541423]
[21]
Hershko A, Heller H, Elias S, Ciechanover A. Components of ubiquitin-protein ligase system. Resolution, affinity purification, and role in protein breakdown. J Biol Chem 1983; 258(13): 8206-14.
[http://dx.doi.org/10.1016/S0021-9258(20)82050-X] [PMID: 6305978]
[22]
Metzger MB, Pruneda JN, Klevit RE, Weissman AM. RING-type E3 ligases: master manipulators of E2 ubiquitin-conjugating enzymes and ubiquitination. Biochim Biophys Acta 2014; 1843(1): 47-60.
[http://dx.doi.org/10.1016/j.bbamcr.2013.05.026] [PMID: 23747565]
[23]
Finley D. Recognition and processing of ubiquitin-protein conjugates by the proteasome. Annu Rev Biochem 2009; 78: 477-513.
[http://dx.doi.org/10.1146/annurev.biochem.78.081507.101607] [PMID: 19489727]
[24]
Lander GC, Estrin E, Matyskiela ME, Bashore C, Nogales E, Martin A. Complete subunit architecture of the proteasome regulatory particle. Nature 2012; 482(7384): 186-91.
[http://dx.doi.org/10.1038/nature10774] [PMID: 22237024]
[25]
Das S, Roth CP, Wasson LM, Vishwanatha JK. Signal transducer and activator of transcription-6 (STAT6) is a constitutively expressed survival factor in human prostate cancer. Prostate 2007; 67(14): 1550-64.
[http://dx.doi.org/10.1002/pros.20640] [PMID: 17705178]
[26]
Amirikia KC, Mills P, Bush J, Newman LA. Higher population-based incidence rates of triple-negative breast cancer among young African-American women: Implications for breast cancer screening recommendations. Cancer 2011; 117(12): 2747-53.
[http://dx.doi.org/10.1002/cncr.25862] [PMID: 21656753]
[27]
Pietras RJ, Márquez-Garbán DC. Membrane-associated estrogen receptor signaling pathways in human cancers. Clin Cancer Res 2007; 13(16): 4672-6.
[http://dx.doi.org/10.1158/1078-0432.CCR-07-1373] [PMID: 17699844]
[28]
Hammes SR, Levin ER. Extranuclear steroid receptors: nature and actions. Endocr Rev 2007; 28(7): 726-41.
[http://dx.doi.org/10.1210/er.2007-0022] [PMID: 17916740]
[29]
Yan Y, Li X, Blanchard A, et al. Expression of both estrogen receptor-beta 1 (ER-β1) and its co-regulator steroid receptor RNA activator protein (SRAP) are predictive for benefit from tamoxifen therapy in patients with estrogen receptor-alpha (ER-α)-negative early breast cancer (EBC). Ann Oncol 2013; 24(8): 1986-93.
[http://dx.doi.org/10.1093/annonc/mdt132] [PMID: 23579816]
[30]
Pezzuto A, Carico E. Role of HIF-1 in Cancer Progression: Novel Insights. A Review. Curr Mol Med 2018; 18(6): 343-51.
[http://dx.doi.org/10.2174/1566524018666181109121849] [PMID: 30411685]
[31]
Thomas C, Rajapaksa G, Nikolos F, et al. ERbeta1 represses basal breast cancer epithelial to mesenchymal transition by destabilizing EGFR. Breast Cancer Res 2012; 14(6): R148.
[http://dx.doi.org/10.1186/bcr3358] [PMID: 23158001]
[32]
Waterman H, Levkowitz G, Alroy I, Yarden Y. The RING finger of c-Cbl mediates desensitization of the epidermal growth factor receptor. J Biol Chem 1999; 274(32): 22151-4.
[http://dx.doi.org/10.1074/jbc.274.32.22151] [PMID: 10428778]
[33]
Jiang X, Huang F, Marusyk A, Sorkin A. Grb2 regulates internalization of EGF receptors through clathrin-coated pits. Mol Biol Cell 2003; 14(3): 858-70.
[http://dx.doi.org/10.1091/mbc.e02-08-0532] [PMID: 12631709]
[34]
Confalonieri S, Salcini AE, Puri C, Tacchetti C, Di Fiore PP. Tyrosine phosphorylation of Eps15 is required for ligand-regulated, but not constitutive, endocytosis. J Cell Biol 2000; 150(4): 905-12.
[http://dx.doi.org/10.1083/jcb.150.4.905] [PMID: 10953014]
[35]
Ramaekers CH, Wouters BG. Regulatory functions of ubiquitin in diverse DNA damage responses. Curr Mol Med 2011; 11(2): 152-69.
[http://dx.doi.org/10.2174/156652411794859269] [PMID: 21342128]
[36]
Lu Q, Hope LW, Brasch M, Reinhard C, Cohen SN. TSG101 interaction with HRS mediates endosomal trafficking and receptor down-regulation. Proc Natl Acad Sci USA 2003; 100(13): 7626-31.
[http://dx.doi.org/10.1073/pnas.0932599100] [PMID: 12802020]
[37]
Bache KG, Brech A, Mehlum A, Stenmark H. Hrs regulates multivesicular body formation via ESCRT recruitment to endosomes. J Cell Biol 2003; 162(3): 435-42.
[http://dx.doi.org/10.1083/jcb.200302131] [PMID: 12900395]
[38]
Radinsky R, Risin S, Fan D, et al. Level and function of epidermal growth factor receptor predict the metastatic potential of human colon carcinoma cells. Clin Cancer Res 1995; 1(1): 19-31.
[PMID: 9815883]
[39]
Harris AL, Nicholson S, Sainsbury R, Wright C, Farndon J. Epidermal growth factor receptor and other oncogenes as prognostic markers. J Natl Cancer Inst Monogr 1992; (11): 181-7.
[PMID: 1627426]
[40]
Jain A, Jain SK. Advances in Tumor Targeted Liposomes. Curr Mol Med 2018; 18(1): 44-57.
[http://dx.doi.org/10.2174/1566524018666180416101522] [PMID: 29663884]
[41]
Price JT, Bonovich MT, Kohn EC. The biochemistry of cancer dissemination. Crit Rev Biochem Mol Biol 1997; 32(3): 175-253.
[http://dx.doi.org/10.3109/10409239709082573] [PMID: 9239493]
[42]
Reis-Filho JS, Pinheiro C, Lambros MB, et al. EGFR amplification and lack of activating mutations in metaplastic breast carcinomas. J Pathol 2006; 209(4): 445-53.
[http://dx.doi.org/10.1002/path.2004] [PMID: 16739104]
[43]
Hoadley KA, Weigman VJ, Fan C, et al. EGFR associated expression profiles vary with breast tumor subtype. BMC Genomics 2007; 8: 258.
[http://dx.doi.org/10.1186/1471-2164-8-258] [PMID: 17663798]
[44]
Burness ML, Grushko TA, Olopade OI. Epidermal growth factor receptor in triple-negative and basal-like breast cancer: promising clinical target or only a marker? Cancer J 2010; 16(1): 23-32.
[http://dx.doi.org/10.1097/PPO.0b013e3181d24fc1] [PMID: 20164687]

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