Quantitative assays for the measurement of HER1-HER2 heterodimerization and phosphorylation in cell lines and breast tumors: Applications for diagnostics and targeted drug mechanism of action

Breast Cancer Research

Volumn 13, Issue 2, 2011, Pages

  DeFazio Eli, Lisa ^a   Strommen, Kristi ^a   Dao Pick, Trang ^a   Parry, Gordon ^a   Goodman, Laurie ^a   Winslow, John ^a  

^a MONOGRAM BIOSCIENCES   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ANTINEOPLASTIC AGENT; EPIDERMAL GROWTH FACTOR; EPIDERMAL GROWTH FACTOR RECEPTOR 1; EPIDERMAL GROWTH FACTOR RECEPTOR 2; ERLOTINIB; HETERODIMER; LAPATINIB; LIGAND; PERTUZUMAB; UNCLASSIFIED DRUG; 2C4 ANTIBODY; EGFR PROTEIN, HUMAN; EPIDERMAL GROWTH FACTOR RECEPTOR; ERBB2 PROTEIN, HUMAN; MONOCLONAL ANTIBODY; PROTEIN KINASE INHIBITOR; QUINAZOLINE DERIVATIVE;

AFFINITY LABELING; ARTICLE; BREAST TUMOR; CAPILLARY ELECTROPHORESIS; CELL LYSATE; CONTROLLED STUDY; DRUG MECHANISM; DRUG TARGETING; FLOW CYTOMETRY; FLUORESCENCE; HUMAN; HUMAN CELL; IMMUNODETECTION; IMMUNOPRECIPITATION; OUTCOME ASSESSMENT; PREVALENCE; PROTEIN ANALYSIS; PROTEIN EXPRESSION; PROTEIN PHOSPHORYLATION; QUANTITATIVE ASSAY; RECEPTOR BINDING; SIGNAL TRANSDUCTION; TISSUE SECTION; TREATMENT RESPONSE; TUMOR CELL LINE; WESTERN BLOTTING; CHEMISTRY; DRUG RESISTANCE; FEMALE; METABOLISM; PHOSPHORYLATION; PROTEIN MULTIMERIZATION; STAINING; TREATMENT OUTCOME;

ANTIBODIES, MONOCLONAL; BREAST NEOPLASMS; CELL LINE, TUMOR; DRUG RESISTANCE, NEOPLASM; EPIDERMAL GROWTH FACTOR; FEMALE; HUMANS; PHOSPHORYLATION; PROTEIN KINASE INHIBITORS; PROTEIN MULTIMERIZATION; QUINAZOLINES; RECEPTOR, EPIDERMAL GROWTH FACTOR; RECEPTOR, ERBB-2; SIGNAL TRANSDUCTION; STAINING AND LABELING; TREATMENT OUTCOME;

EID: 79955088678     PISSN: 14655411     EISSN: 1465542X     Source Type: Journal    
DOI: 10.1186/bcr2866     Document Type: Article

References (73)

1. Cochet C, Kashles O, Chambaz EM, Borrello I, King CR, Schlessinger J. Demonstration of epidermal growth factor-induced receptor dimerization in living cells using a chemical covalent cross-linking agent. J Biol Chem 1988, 263:3290-3295.
2. Heldin CH. Dimerization of cell surface receptors in signal transduction. Cell 1995, 80:213-223. 10.1016/0092-8674(95)90404-2, 7834741.
3. Greenfield C, Hiles I, Waterfield MD, Federwisch M, Wollmer A, Blundell TL, McDonald N. Epidermal growth factor binding induces a conformational change in the external domain of its receptor. EMBO J 1989, 8:4115-4123. 401594, 2591370.
4. Higashiyama S, Abraham JA, Miller J, Fiddes JC, Klagsbrun M. A heparin-binding growth factor secreted by macrophage-like cells that is related to EGF. Science 1991, 251:936-939. 10.1126/science.1840698, 1840698.
5. Shing Y, Christofori G, Hanahan D, Ono Y, Sasada R, Igarashi K, Folkman J. Betacellulin: a mitogen from pancreatic beta cell tumors. Science 1993, 259:1604-1607. 10.1126/science.8456283, 8456283.
6. Shoyab M, Plowman GD, McDonald VL, Bradley JG, Todaro GJ. Structure and function of human amphiregulin: a member of the epidermal growth factor family. Science 1989, 243:1074-1076. 10.1126/science.2466334, 2466334.
7. Toyoda H, Komurasaki T, Uchida D, Takayama Y, Isobe T, Okuyama T, Hanada K. Epiregulin. A novel epidermal growth factor with mitogenic activity for rat primary hepatocytes. J Biol Chem 1995, 270:7495-7500. 10.1074/jbc.270.13.7495, 7706296.
8. Di Fiore PP, Pierce JH, Kraus MH, Segatto O, King CR, Aaronson SA. erbB-2 is a potent oncogene when overexpressed in NIH/3T3 cells. Science 1987, 237:178-182. 10.1126/science.2885917, 2885917.
9. Yarden Y, Sliwkowski MX. Untangling the ErbB signalling network. Nat Rev Mol Cell Biol 2001, 2:127-137. 10.1038/35052073, 11252954.
10. Gan HK, Walker F, Burgess AW, Rigopoulos A, Scott AM, Johns TG. The epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor AG1478 increases the formation of inactive untethered EGFR dimers. Implications for combination therapy with monoclonal antibody 806. J Biol Chem 2007, 282:2840-2850.
11. Yu X, Sharma KD, Takahashi T, Iwamoto R, Mekada E. Ligand-independent dimer formation of epidermal growth factor receptor (EGFR) is a step separable from ligand-induced EGFR signaling. Mol Biol Cell 2002, 13:2547-2557. 10.1091/mbc.01-08-0411, 117333, 12134089.
12. Liu P, Sudhaharan T, Koh RM, Hwang LC, Ahmed S, Maruyama IN, Wohland T. Investigation of the dimerization of proteins from the epidermal growth factor receptor family by single wavelength fluorescence cross-correlation spectroscopy. Biophys J 2007, 93:684-698. 10.1529/biophysj.106.102087, 1896234, 17468161.
13. Sako Y, Minoghchi S, Yanagida T. Single-molecule imaging of EGFR signalling on the surface of living cells. Nat Cell Biol 2000, 2:168-172. 10.1038/35004044, 10707088.
14. Tao RH, Maruyama IN. All EGF(ErbB) receptors have preformed homo- and heterodimeric structures in living cells. J Cell Sci 2008, 121:3207-3217. 10.1242/jcs.033399, 18782861.
15. Klapper LN, Kirschbaum MH, Sela M, Yarden Y. Biochemical and clinical implications of the ErbB/HER signaling network of growth factor receptors. Adv Cancer Res 2000, 77:25-79.
16. Carlson RW, Moench SJ, Hammond ME, Perez EA, Burstein HJ, Allred DC, Vogel CL, Goldstein LJ, Somlo G, Gradishar WJ, Hudis CA, Jahanzeb M, Stark A, Wolff AC, Press MF, Winer EP, Paik S, Ljung BM. HER2 testing in breast cancer: NCCN Task Force report and recommendations. J Natl Compr Canc Netw 2006, 4(S1-22):S23-24. quiz.
17. Hudis CA. Trastuzumab--mechanism of action and use in clinical practice. N Engl J Med 2007, 357:39-51. 10.1056/NEJMra043186, 17611206.
18. Ekstrand AJ, Sugawa N, James CD, Collins VP. Amplified and rearranged epidermal growth factor receptor genes in human glioblastomas reveal deletions of sequences encoding portions of the N- and/or C-terminal tails. Proc Natl Acad Sci USA 1992, 89:4309-4313. 10.1073/pnas.89.10.4309, 49071, 1584765.
19. Valverius EM, Bates SE, Stampfer MR, Clark R, McCormick F, Salomon DS, Lippman ME, Dickson RB. Transforming growth factor alpha production and epidermal growth factor receptor expression in normal and oncogene transformed human mammary epithelial cells. Mol Endocrinol 1989, 3:203-214. 10.1210/mend-3-1-203, 2783757.
20. Muller WJ, Arteaga CL, Muthuswamy SK, Siegel PM, Webster MA, Cardiff RD, Meise KS, Li F, Halter SA, Coffey RJ. Synergistic interaction of the Neu proto-oncogene product and transforming growth factor alpha in the mammary epithelium of transgenic mice. Mol Cell Biol 1996, 16:5726-5736. 231573, 8816486.
21. DiGiovanna MP, Lerman MA, Coffey RJ, Muller WJ, Cardiff RD, Stern DF. Active signaling by Neu in transgenic mice. Oncogene 1998, 17:1877-1884. 10.1038/sj.onc.1202091, 9778054.
22. Nieto Y, Nawaz F, Jones RB, Shpall EJ, Nawaz S. Prognostic significance of overexpression and phosphorylation of epidermal growth factor receptor (EGFR) and the presence of truncated EGFRvIII in locoregionally advanced breast cancer. J Clin Oncol 2007, 25:4405-4413. 10.1200/JCO.2006.09.8822, 17906204.
23. Simpson BJ, Phillips HA, Lessells AM, Langdon SP, Miller WR. c-erbB growth-factor-receptor proteins in ovarian tumours. Int J Cancer 1995, 64:202-206. 10.1002/ijc.2910640310, 7622309.
24. DiGiovanna MP, Stern DF, Edgerton SM, Whalen SG, Moore D, Thor AD. Relationship of epidermal growth factor receptor expression to ErbB-2 signaling activity and prognosis in breast cancer patients. J Clin Oncol 2005, 23:1152-1160. 10.1200/JCO.2005.09.055, 15718311.
25. Arteaga CL, O'Neill A, Moulder SL, Pins M, Sparano JA, Sledge GW, Davidson NE. A phase I-II study of combined blockade of the ErbB receptor network with trastuzumab and gefitinib in patients with HER2 (ErbB2)-overexpressing metastatic breast cancer. Clin Cancer Res 2008, 14:6277-6283. 10.1158/1078-0432.CCR-08-0482, 2925197, 18829509.
26. de Alava E, Ocana A, Abad M, Montero JC, Esparis-Ogando A, Rodriguez CA, Otero AP, Hernandez T, Cruz JJ, Pandiella A. Neuregulin expression modulates clinical response to trastuzumab in patients with metastatic breast cancer. J Clin Oncol 2007, 25:2656-2663. 10.1200/JCO.2006.08.6850, 17602072.
27. Piccart M, Lohrisch C, Di Leo A, Larsimont D. The predictive value of HER2 in breast cancer. Oncology 2001, 61(Suppl 2):73-82.
28. Goodman LJ, Weston JK, Mukherjee A, Sperinde J, Paquet AC, Williams S, Parry G, Bates M, Koestler W, Lipton A. Quantitative measurement of HER3 total protein (H3T) and association with clinical outcome in HER2 positive metastatic breast cancer patients treated with trastuzumab. In 2009 ASCO Annual Meeting. J Clin Oncol 2009, 27:15s.
29. Sperinde J, Jin X, Banerjee J, Penuel E, Saha A, Diedrich G, Huang W, Leitzel K, Weidler J, Ali SM, Fuchs EM, Singer CF, Kostler WJ, Bates M, Parry G, Winslow J, Lipton A. Quantitation of p95HER2 in Paraffin sections by using a p95-specific antibody and correlation with outcome in a cohort of trastuzumab-treated breast cancer patients. Clin Cancer Res 2010, 16:4226-4235. 10.1158/1078-0432.CCR-10-0410, 20664024.
30. Ritter CA, Perez-Torres M, Rinehart C, Guix M, Dugger T, Engelman JA, Arteaga CL. Human breast cancer cells selected for resistance to trastuzumab in vivo overexpress epidermal growth factor receptor and ErbB ligands and remain dependent on the ErbB receptor network. Clin Cancer Res 2007, 13:4909-4919. 10.1158/1078-0432.CCR-07-0701, 17699871.
31. Dua R, Zhang J, Nhonthachit P, Penuel E, Petropoulos C, Parry G. EGFR over-expression and activation in high HER2, ER negative breast cancer cell line induces trastuzumab resistance. Breast Cancer Res Treat 2010, 122:685-697. 10.1007/s10549-009-0592-x, 19859802.
32. Nicholson RI, Hutcheson IR, Harper ME, Knowlden JM, Barrow D, McClelland RA, Jones HE, Wakeling AE, Gee JM. Modulation of epidermal growth factor receptor in endocrine-resistant, oestrogen receptor-positive breast cancer. Endocr Relat Cancer 2001, 8:175-182. 10.1677/erc.0.0080175, 11566608.
33. Nicholson RI, McClelland RA, Gee JM, Manning DL, Cannon P, Robertson JF, Ellis IO, Blamey RW. Epidermal growth factor receptor expression in breast cancer: association with response to endocrine therapy. Breast Cancer Res Treat 1994, 29:117-125. 10.1007/BF00666187, 7912565.
34. Knowlden JM, Hutcheson IR, Jones HE, Madden T, Gee JM, Harper ME, Barrow D, Wakeling AE, Nicholson RI. Elevated levels of epidermal growth factor receptor/c-erbB2 heterodimers mediate an autocrine growth regulatory pathway in tamoxifen-resistant MCF-7 cells. Endocrinology 2003, 144:1032-1044. 10.1210/en.2002-220620, 12586780.
35. Gajadhar A, Guha A. A proximity ligation assay using transiently transfected, epitope-tagged proteins: application for in situ detection of dimerized receptor tyrosine kinases. Biotechniques 2010, 48:145-152. 10.2144/000113354, 20359299.
36. Leuchowius KJ, Weibrecht I, Landegren U, Gedda L, Soderberg O. Flow cytometric in situ proximity ligation analyses of protein interactions and post-translational modification of the epidermal growth factor receptor family. Cytometry A 2009, 75:833-839.
37. Spears M, Bartlett JM. Human epidermal growth factor receptor dimerization analysis in breast cancer diagnosis: potential for improving testing accuracy and treatment selection. Mol Diagn Ther 2009, 13:359-365.
38. Shi Y, Huang W, Tan Y, Jin X, Dua R, Penuel E, Mukherjee A, Sperinde J, Pannu H, Chenna A, DeFazio-Eli L, Pidaparthi S, Badal Y, Wallweber G, Chen L, Williams S, Tahir H, Larson J, Goodman L, Whitcomb J, Petropoulos C, Winslow J. A novel proximity assay for the detection of proteins and protein complexes: quantitation of HER1 and HER2 total protein expression and homodimerization in formalin-fixed, paraffin-embedded cell lines and breast cancer tissue. Diagn Mol Pathol 2009, 18:11-21. 10.1097/PDM.0b013e31818cbdb2, 19214113.
39. Larson JS, Goodman LJ, Tan Y, DeFazio-Eli L, Paquet AC, Cook JW, Rivera A, Frankson K, Bose J, Chen L, Cheung J, Shi Y, Irwin S, Kiss LDB, Huang W, Utter S, Sherwood T, Bates M, Weidler J, Parry G, Winslow J, Petropoulos CJ, Whitcomb J. Analytical validation of a highly sensitive, accurate, and reproducible assay (HERmark) for the measurement of HER2 total protein and HER2 homodimers in FFPE breast cancer tumor specimens. Pathol Res Int 2010, 2010::814176. 2990097, 21151530.
40. Stommel JM, Kimmelman AC, Ying H, Nabioullin R, Ponugoti AH, Wiedemeyer R, Stegh AH, Bradner JE, Ligon KL, Brennan C, Chin L, DePinho RA. Coactivation of receptor tyrosine kinases affects the response of tumor cells to targeted therapies. Science 2007, 318:287-290. 10.1126/science.1142946, 17872411.
41. Lee-Hoeflich ST, Crocker L, Yao E, Pham T, Munroe X, Hoeflich KP, Sliwkowski MX, Stern HM. A central role for HER3 in HER2-amplified breast cancer: implications for targeted therapy. Cancer Res 2008, 68:5878-5887. 10.1158/0008-5472.CAN-08-0380, 18632642.
42. Junttila TT, Akita RW, Parsons K, Fields C, Lewis GD, Friedman LS, Sampath D, Sliwkowski MX. Ligand-independent HER2/HER3/PI3K complex is disrupted by trastuzumab and is effectively inhibited by the PI3K inhibitor GDC-0941. Cancer Cell 2009, 15:429-440. 10.1016/j.ccr.2009.03.020, 19411071.
43. Franklin MC, Carey KD, Vajdos FF, Leahy DJ, de Vos AM, Sliwkowski MX. Insights into ErbB signaling from the structure of the ErbB2-pertuzumab complex. Cancer Cell 2004, 5:317-328. 10.1016/S1535-6108(04)00083-2, 15093539.
44. Hughes JB, Berger C, Rodland MS, Hasmann M, Stang E, Madshus IH. Pertuzumab increases epidermal growth factor receptor down-regulation by counteracting epidermal growth factor receptor-ErbB2 heterodimerization. Mol Cancer Ther 2009, 8:1885-1892. 10.1158/1535-7163.MCT-09-0291, 19584234.
45. Takai N, Jain A, Kawamata N, Popoviciu LM, Said JW, Whittaker S, Miyakawa I, Agus DB, Koeffler HP. 2C4, a monoclonal antibody against HER2, disrupts the HER kinase signaling pathway and inhibits ovarian carcinoma cell growth. Cancer 2005, 104:2701-2708. 10.1002/cncr.21533, 16265675.
46. Anido J, Matar P, Albanell J, Guzman M, Rojo F, Arribas J, Averbuch S, Baselga J. ZD1839, a specific epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, induces the formation of inactive EGFR/HER2 and EGFR/HER3 heterodimers and prevents heregulin signaling in HER2-overexpressing breast cancer cells. Clin Cancer Res 2003, 9:1274-1283.
47. Scaltriti M, Verma C, Guzman M, Jimenez J, Parra JL, Pedersen K, Smith DJ, Landolfi S, Ramon YCS, Arribas J, Baselga J. Lapatinib, a HER2 tyrosine kinase inhibitor, induces stabilization and accumulation of HER2 and potentiates trastuzumab-dependent cell cytotoxicity. Oncogene 2008, 28:803-814.
48. Arteaga CL, Ramsey TT, Shawver LK, Guyer CA. Unliganded epidermal growth factor receptor dimerization induced by direct interaction of quinazolines with the ATP binding site. J Biol Chem 1997, 272:23247-23254. 10.1074/jbc.272.37.23247, 9287333.
49. Allred DC, Harvey JM, Berardo M, Clark GM. Prognostic and predictive factors in breast cancer by immunohistochemical analysis. Mod Pathol 1998, 11:155-168.
50. Press MF, Finn RS, Cameron D, Di Leo A, Geyer CE, Villalobos IE, Santiago A, Guzman R, Gasparyan A, Ma Y, Danenberg K, Martin AM, Williams L, Oliva C, Stein S, Gagnon R, Arbushites M, Koehler MT. HER-2 gene amplification, HER-2 and epidermal growth factor receptor mRNA and protein expression, and lapatinib efficacy in women with metastatic breast cancer. Clin Cancer Res 2008, 14:7861-7870. 10.1158/1078-0432.CCR-08-1056, 19047115.
51. Yao E, Zhou W, Lee-Hoeflich ST, Truong T, Haverty PM, Eastham-Anderson J, Lewin-Koh N, Gunter B, Belvin M, Murray LJ, Friedman LS, Sliwkowski MX, Hoeflich KP. Suppression of HER2/HER3-mediated growth of breast cancer cells with combinations of GDC-0941 PI3K inhibitor, trastuzumab, and pertuzumab. Clin Cancer Res 2009, 15:4147-4156. 10.1158/1078-0432.CCR-08-2814, 19509167.
52. Adams CW, Allison DE, Flagella K, Presta L, Clarke J, Dybdal N, McKeever K, Sliwkowski MX. Humanization of a recombinant monoclonal antibody to produce a therapeutic HER dimerization inhibitor, pertuzumab. Cancer Immunol Immunother 2006, 55:717-727. 10.1007/s00262-005-0058-x, 16151804.
53. Agus DB, Akita RW, Fox WD, Lewis GD, Higgins B, Pisacane PI, Lofgren JA, Tindell C, Evans DP, Maiese K, Scher HI, Sliwkowski MX. Targeting ligand-activated ErbB2 signaling inhibits breast and prostate tumor growth. Cancer Cell 2002, 2:127-137. 10.1016/S1535-6108(02)00097-1, 12204533.
54. Cai Z, Zhang G, Zhou Z, Bembas K, Drebin JA, Greene MI, Zhang H. Differential binding patterns of monoclonal antibody 2C4 to the ErbB3-p185(her2/neu) and the EGFR-p185(her2/neu) complexes. Oncogene 2008, 11:11. 2819401, 18264138.
55. Wood ER, Truesdale AT, McDonald OB, Yuan D, Hassell A, Dickerson SH, Ellis B, Pennisi C, Horne E, Lackey K, Alligood KJ, Rusnak DW, Gilmer TM, Shewchuck L. A unique structure for epidermal growth factor receptor bound to GW572016 (Lapatinib): relationships among protein conformation, inhibitor off-rate, and receptor activity in tumor cells. Cancer Res 2004, 64:6652-6659. 10.1158/0008-5472.CAN-04-1168, 15374980.
56. Wakeling AE, Guy SP, Woodburn JR, Ashton SE, Curry BJ, Barker AJ, Gibson KH. ZD1839 (Iressa): an orally active inhibitor of epidermal growth factor signaling with potential for cancer therapy. Cancer Res 2002, 62:5749-5754.
57. Schaefer G, Shao L, Totpal K, Akita RW. Erlotinib directly inhibits HER2 kinase activation and downstream signaling events in intact cells lacking epidermal growth factor receptor expression. Cancer Res 2007, 67:1228-1238. 10.1158/0008-5472.CAN-06-3493, 17283159.
58. Stamos J, Sliwkowski MX, Eigenbrot C. Structure of the epidermal growth factor receptor kinase domain alone and in complex with a 4-anilinoquinazoline inhibitor. J Biol Chem 2002, 277:46265-46272. 10.1074/jbc.M207135200, 12196540.
59. Cicenas J, Urban P, Kung W, Vuaroqueaux V, Labuhn M, Wight E, Eppenberger U, Eppenberger-Castori S. Phosphorylation of tyrosine 1248-ERBB2 measured by chemiluminescence-linked immunoassay is an independent predictor of poor prognosis in primary breast cancer patients. Eur J Cancer 2006, 42:636-645. 10.1016/j.ejca.2005.11.012, 16414259.
60. Modi S, DiGiovanna MP, Lu Z, Moskowitz C, Panageas KS, Van Poznak C, Hudis CA, Norton L, Tan L, Stern DF, Carter D, Seidman AD. Phosphorylated/activated HER2 as a marker of clinical resistance to single agent taxane chemotherapy for metastatic breast cancer. Cancer Invest 2005, 23:483-487. 10.1080/07357900500201301, 16203655.
61. Frogne T, Laenkholm AV, Lyng MB, Henriksen KL, Lykkesfeldt AE. Determination of HER2 phosphorylation at tyrosine 1221/1222 improves prediction of poor survival for breast cancer patients with hormone receptor-positive tumors. Breast Cancer Res 2009, 11:R11. 10.1186/bcr2230, 2687716, 19239686.
62. Thor AD, Liu S, Edgerton S, Moore D, Kasowitz KM, Benz CC, Stern DF, DiGiovanna MP. Activation (tyrosine phosphorylation) of ErbB-2 (HER-2/neu): a study of incidence and correlation with outcome in breast cancer. J Clin Oncol 2000, 18:3230-3239.
63. Johnston S, Trudeau M, Kaufman B, Boussen H, Blackwell K, LoRusso P, Lombardi DP, Ben Ahmed S, Citrin DL, DeSilvio ML, Harris J, Westlund RE, Salazar V, Zaks TZ, Spector NL. Phase II study of predictive biomarker profiles for response targeting human epidermal growth factor receptor 2 (HER-2) in advanced inflammatory breast cancer with lapatinib monotherapy. J Clin Oncol 2008, 26:1066-1072. 10.1200/JCO.2007.13.9949, 18212337.
64. Wulfkuhle J, Pierobon M, Laird J, Espina I, Liotta L, Esserman L, Petricoin E. Discovery of a new phospho-HER2+/FISH- molecular subtype of human breast cancer by functional pathway mapping. In 2009 ASCO Annual Meeting, J Clin Oncol 27:15s.
65. Hijiya N, Miyawaki M, Kawahara K, Akamine S, Tsuji K, Kadota J, Akizuki S, Uchida T, Matsuura K, Tsukamoto Y, Moriyama M. Phosphorylation status of epidermal growth factor receptor is closely associated with responsiveness to gefitinib in pulmonary adenocarcinoma. Hum Pathol 2008, 39:316-323. 10.1016/j.humpath.2007.10.027, 18261621.
66. Kuwai T, Nakamura T, Sasaki T, Kim SJ, Fan D, Villares GJ, Zigler M, Wang H, Bar-Eli M, Kerbel RS, Fidler IJ. Phosphorylated epidermal growth factor receptor on tumor-associated endothelial cells is a primary target for therapy with tyrosine kinase inhibitors. Neoplasia 2008, 10:489-500. 2373870, 18472966.
67. Guo L, Abraham J, Flynn DC, Castranova V, Shi X, Qian Y. Individualized survival and treatment response predictions for breast cancers using phospho-EGFR, phospho-ER, phospho-HER2/neu, phospho-IGF-IR/In, phospho-MAPK, and phospho-p70S6K proteins. Int J Biol Markers 2007, 22:1-11.
68. Ono M, Kuwano M. Molecular mechanisms of epidermal growth factor receptor (EGFR) activation and response to gefitinib and other EGFR-targeting drugs. Clin Cancer Res 2006, 12:7242-7251. 10.1158/1078-0432.CCR-06-0646, 17189395.
69. Corkery B, Crown J, Clynes M, O'Donovan N. Epidermal growth factor receptor as a potential therapeutic target in triple-negative breast cancer. Ann Oncol 2009, 20:862-867. 10.1093/annonc/mdn710, 19150933.
70. Hama T, Yuza Y, Saito Y, Ou J, Kondo S, Okabe M, Yamada H, Kato T, Moriyama H, Kurihara S, Urashima M. Prognostic significance of epidermal growth factor receptor phosphorylation and mutation in head and neck squamous cell carcinoma. Oncologist 2009, 14:900-908. 10.1634/theoncologist.2009-0058, 19726454.
71. Emery IF, Battelli C, Auclair PL, Carrier K, Hayes DM. Response to gefitinib and erlotinib in Non-small cell lung cancer: a restrospective study. BMC Cancer 2009, 9:333. 10.1186/1471-2407-9-333, 2758901, 19765296.
72. Mukherjee A, Badal Y, Nguyen X-T, Miller J, Chenna A, Tahir H, Newton A, Parry G, Williams S. Profiling the HER3/PI3K pathway in breast tumors using proximity-directed assays identifies correlations between protein complexes and phosphoproteins. PloS ONE 2011, 6:e16443. 10.1371/journal.pone.0016443, 3030586, 21297994.
73. Pinhel IF, MacNeill FA, Hills MJ, Salter J, Detre S, A'Hern R, Nerurkar A, Osin P, Smith IE, Dowsett M. Extreme loss of immunoreactive p-Akt and p-Erk1/2 during routine fixation of primary breast cancer. Breast Cancer Res 2010, 12:R76. 10.1186/bcr2719, 3005722, 20920193.