Suppressive impact of metronomic chemotherapy using UFT and/or cyclophosphamide on mediators of breast cancer dissemination and invasion

English version

Autoři: Raquel Muñoz ^aff001; Denise Hileeto ^aff002; William Cruz-Muñoz ^aff001; Geoffrey A. Wood ^aff003; Ping Xu ^aff001; Shan Man ^aff001; Alicia Viloria-Petit ^aff004; Robert S. Kerbel ^aff001
Působiště autorů: Biological Sciences Platform, Sunnybrook Research Institute, Toronto, Canada ^aff001; School of Optometry & Vision Science, University of Waterloo, Waterloo, Ontario, Canada ^aff002; Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph, Ontario, Canada ^aff003; Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, Guelph, Ontario, Canada ^aff004; Department of Medical Biophysics, University of Toronto, Toronto, Canada ^aff005
Vyšlo v časopise: PLoS ONE 14(9)
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pone.0222580

Souhrn

Metronomic chemotherapy using the 5-FU prodrug uracil-tegafur (UFT) and cyclophosphamide (CTX) was previously shown to only modestly delay primary tumor growth, but nevertheless markedly suppressed the development of micro-metastasis in an orthotopic breast cancer xenograft model, using the metastatic variant of the MDA-MB-231 cell line, 231/LM2-4. Furthermore, a remarkable prolongation of survival, with no toxicity, was observed in a model of postsurgical advanced metastatic disease. A question that has remained unanswered is the seemingly selective anti-metastatic mechanisms of action responsible for this treatment. We assessed the in vivo effect of metronomic UFT, CTX or their combination, on vascular density, collagen deposition and c-Met (cell mediators or modulators of tumor cell invasion or dissemination) via histochemistry/immunohistochemistry of primary tumor sections. We also assessed the effect of continuous exposure to low and non-toxic doses of active drug metabolites 5-fluorouracil (5-FU), 4-hydroperoxycyclophosphamide (4-HC) or their combination, on 231/LM2-4 cell invasiveness in vitro. In the in vivo studies, a significant reduction in vascular density and p-Met[Y1003] levels was associated with UFT+CTX treatment. All treatments reduced intratumoral collagen deposition. In the in vitro studies, a significant reduction of collagen IV invasion by all treatments was observed. The 3D structures formed by 231/LM2-4 on Matrigel showed a predominantly Mass phenotype under treated conditions and Stellate phenotype in untreated cultures. Taken together, the results suggest the low-dose metronomic chemotherapy regimens tested can suppress several mediators of tumor invasiveness highlighting a new perspective for the anti-metastatic efficacy of metronomic chemotherapy.

Klíčová slova:

Biology and life sciences – Biochemistry – Research and analysis methods – Proteins – Medicine and health sciences – Pathology and laboratory medicine – Diagnostic medicine – Signs and symptoms – Clinical medicine – Pharmaceutics – Drug therapy – Oncology – Cancer treatment – Cancers and neoplasms – Collagens – Specimen preparation and treatment – Staining – Necrosis – Cancer chemotherapy – Clinical oncology – Chemotherapy – Nuclear staining – Breast tumors – Breast cancer – Invasive tumors – Cytoplasmic staining

Zdroje

1. Browder T, Butterfield CE, Kraling BM, Marshall B, O'Reilly MS, Folkman J. Antiangiogenic scheduling of chemotherapy improves efficacy against experimental drug-resistant cancer. Cancer Res. 2000;60:1878–1886. 10766175

2. Klement G, Baruchel S, Rak J, Man S, Clark K, Hicklin D, et al. Continuous low-dose therapy with vinblastine and VEGF receptor-2 antibody induces sustained tumor regression without overt toxicity. J Clin Invest. 2000 Apr 15;105(8): R15–R24. doi: 10.1172/JCI8829 10772661

3. Hanahan D, Bergers G, Bergsland E. Less is more, regularly: metronomic dosing of cytotoxic drugs can target tumor angiogenesis in mice. J Clin Invest. 2000;105:1045–1047. doi: 10.1172/JCI9872 10772648

4. Pasquier E, Kavallaris M, Andre N. Metronomic chemotherapy: new rationale for new directions. Nat Rev Clin Oncol. 2010;7:455–465. doi: 10.1038/nrclinonc.2010.82 20531380

5. Bocci G, Tuccori M, Emmenegger U, Liguori V, Falcone A, Kerbel RS, et al. Cyclophosphamide-methotrexate 'metronomic' chemotherapy for the palliative treatment of metastatic breast cancer. A comparative pharmacoeconomic evaluation. Ann Oncol. 2005;16:1243–1252. doi: 10.1093/annonc/mdi240 15905308

6. Kerbel RS, Kamen BA. Antiangiogenic basis of low-dose metronomic chemotherapy. Nat Rev Cancer. 2004;4:423–436. doi: 10.1038/nrc1369 15170445

7. Ghiringhelli F, Menard C, Puig PE, Ladoire S, Roux S, Martin F, et al. Metronomic cyclophosphamide regimen selectively depletes CD4+CD25+ regulatory T cells and restores T and NK effector functions in end stage cancer patients. Cancer Immunol Immunother. 2007;56:641–648. doi: 10.1007/s00262-006-0225-8 16960692

8. Shaked Y, Pham E, Hariharan S, Magidey K, Beyar-Katz O, Xu P, et al. Evidence implicating immunological host effects in the efficacy of metronomic low-dose chemotherapy. Cancer Res. 2016;76:5983–5993. doi: 10.1158/0008-5472.CAN-16-0136 27569209

9. Wu J, Jordan M, Waxman DJ. Metronomic cyclophosphamide activation of anti-tumor immunity: tumor model, mouse host, and drug schedule dependence of gene responses and their upstream regulators. BMC Cancer. 2016 Aug 11;16:623. doi: 10.1186/s12885-016-2597-2 27515027

10. Wu J, Waxman DJ. Metronomic cyclophosphamide schedule-dependence of innate immune cell recruitment and tumor regression in an implanted glioma model. Cancer Lett. 2014;353:272–280. doi: 10.1016/j.canlet.2014.07.033 25069038

11. Ferrer-Font L, Arias-Ramos N, Lope-Piedrafita S, Julia-Sape M, Pumarola M, Arus C, et al. Metronomic treatment in immunocompetent preclinical GL261 glioblastoma: effects of cyclophosphamide and temozolomide. NMR Biomed. 2017 Sep;30(9):e3748.

12. Andre N, Tsai K, Carre M, Pasquier E. Metronomic Chemotherapy: Direct Targeting of Cancer Cells after all? Trends Cancer. 2017;3:319–325. doi: 10.1016/j.trecan.2017.03.011 28718409

13. Munoz R, Man S, Shaked Y, Lee C, Wong J, Francia G, et al. Highly efficacious non-toxic treatment for advanced metastatic breast cancer using combination UFT-cyclophosphamide metronomic chemotherapy. Cancer Res. 2006;66:3386–3391. doi: 10.1158/0008-5472.CAN-05-4411 16585158

14. Jedeszko C, Paez-Ribes M, Di Desidero T, Bocci G, Man S, Lee CR, et al. Orthotopic primary and postsurgical adjuvant or metastatic renal cell carcinoma therapy models reveal potent anti-tumor activity of minimally toxic metronomic oral topotecan with pazopanib. Sci Transl Med. 2015 Apr 8;7(282):282ra250.

15. Hackl C, Man S, Francia G, Xu P, Kerbel RS. Metronomic oral topotecan prolongs survival and reduces liver metastasis in improved preclinical orthotopic and adjuvant therapy colon cancer models. Gut. 2013;62:259–271. doi: 10.1136/gutjnl-2011-301585 22543158

16. Man S, Bocci G, Francia G, Green S, Jothy S, Bergers G, et al. Antitumor and anti-angiogenic effects in mice of low-dose (metronomic) cyclophosphamide administered continuously through the drinking water. Cancer Res. 2002;62:2731–2735. 12019144

17. Cazzaniga ME, Cortesi L, Ferzi A, Scaltriti L, Cicchiello F, Ciccarese M, et al. Metronomic Chemotherapy in Triple-Negative Metastatic Breast Cancer: The Future Is Now? Int J Breast Cancer 2017 Dec 3;1683060. doi: 10.1155/2017/1683060 29333297

18. Simkens LHJ, van Tinteren H, May A, Ten Tije AJ, Creemers G-JM, Loosveld OJL. et al. Maintenance treatment with capecitabine and bevacizumab in metastatic colorectal cancer, the phase 3 CAIRO3 study of the Dutch Colorectal Cancer Group (DCCG). Lancet. 2015;385:1843–1852. doi: 10.1016/S0140-6736(14)62004-3 25862517

19. Hashimoto K, Man S, Xu P, Cruz-Munoz W, Tang T, Kumar R. et al. Potent preclinical impact of metronomic low-dose oral topotecan combined with the antiangiogenic drug pazopanib for the treatment of ovarian cancer. Mol Cancer Ther. 2010;9:996–1006. doi: 10.1158/1535-7163.MCT-09-0960 20371722

20. Bisogno G, De Salvo GL, Bergeron C, Jenney M, Merks JHM, Minard-Colin V, et al. Maintenance low-dose chemotherapy in patients with high-risk (HR) rhabdomyosarcoma (RMS): A report from the European Paediatric Soft Tissue Sarcoma Study Group (EpSSG). J Clin Oncol. 2018;36 Suppl 18:LBA2–LBA2.

21. Colleoni M, Gray KP, Gelber S, Lang I, Thurlimann B, Gianni L, et al. Low-dose oral cyclophosphamide and methotrexate maintenance for hormone receptor-negative early breast cancer: International Breast Cancer Study Group Trial 22–00. J Clin Oncol. 2016;34:3400–3408. doi: 10.1200/JCO.2015.65.6595 27325862

22. Cazzaniga ME, Dionisio MR, Riva F. Metronomic chemotherapy for advanced breast cancer patients. Cancer Lett. 2017;400:252–258. doi: 10.1016/j.canlet.2016.12.019 28017894

23. Munzone E, Colleoni M. Clinical overview of metronomic chemotherapy in breast cancer. Nat Rev Clin Oncol. 2015;385:1843–1852.

24. Liu Y, Gu F, Liang J, Dai X, Wan C, Hong X, et al. The efficacy and toxicity profile of metronomic chemotherapy for metastatic breast cancer: A meta-analysis. PLoS One. 2017 Mar 15;12(3):e0173693. doi: 10.1371/journal.pone.0173693 28296916

25. Cazzaniga ME, Cortesi L, Ferzi A, Scaltriti L, Cicchiello F, Ciccarese M, et al. Metronomic chemotherapy with oral vinorelbine (mVNR) and capecitabine (mCAPE) in advanced HER2-negative breast cancer patients: is it a way to optimize disease control? Final results of the VICTOR-2 study. Breast Cancer Res Treat. 2016;160:501–509. doi: 10.1007/s10549-016-4009-3 27752847

26. Pasini F, Barile C, Caruso D, Modena Y, Fraccon AP, Bertolaso L, et al. Oral Metronomic Vinorelbine (OMV) in elderly or pretreated patients with advanced non small cell lung cancer: outcome and pharmacokinetics in the real world. Invest New Drugs. 2018;36:927–932. doi: 10.1007/s10637-018-0631-8 29956056

27. Montagna E, Bagnardi V, Cancello G, Sangalli C, Pagan E, Iorfida M, et al. Metronomic Chemotherapy for First-Line Treatment of Metastatic Triple-Negative Breast Cancer: A Phase II Trial. Breast Care (Basel). 2018;13:177–181.

28. Noguchi S, Koyama H, Uchino J, Abe R, Miura S, Sugimachi K, et al. Postoperative adjuvant therapy with tamoxifen, tegafur plus uracil, or both in women with node-negative breast cancer: a pooled analysis of six randomized controlled trials. J Clin Oncol. 2005;23:2171–2184.

29. Kurebayashi J, Nukatsuka M, Fujioka A, Saito H, Takeda S, Unemi N, et al. Postsurgical oral administration of uracil and tegafur inhibits progression of micrometastasis of human breast cancer cells in nude mice. Clin Cancer Res. 1997;3:653–659. 9815733

30. Nakanishi H, Kobayashi K, Nishimura T, Inada K, Tsukamoto T, Tatematsu M. Chemosensitivity of micrometastases and circulating tumour cells to uracil and tegafur as evaluated using LacZ gene-tagged Lewis lung carcinoma cell. Cancer Lett. 1999;142:31–41. doi: 10.1016/s0304-3835(99)00114-7 10424778

31. Uchida J, Okabe H, Nakano K, Fujioka A, Saito H, Sugimoto Y, et al. Contrasting effects of extended low dose versus standard dose shorter course UFT chemotherapy on microscopic versus macroscopic established tumours: implications for optimal postoperative adjuvant chemotherapy. Oncol Rep. 2007;18:313–319. 17611650

32. Bertolini F, Paul S, Mancuso P, Monestiroli S, Gobbi A, Shaked Y, et al. Maximun tolerable dose and low-dose metronomic chemotherapy have opposite effects on the mobilization and viability of circulating endotelial progenitor cells. Cancer Res. 2003;63:4342–4346. 12907602

33. Basaki Y, Chikahisa L, Aoyagi K, Miyadera K, Yonekura K, Hashimoto A, et al. gamma-Hydroxybutyric acid and 5-fluorouracil, metabolites of UFT, inhibit the angiogenesis induced by vascular endothelial growth factor. Angiogenesis. 2001;4:163–73. 11911014

34. Yonekura K, Basaki Y, Chikahisa L, Okabe S, Hashimoto A, Miyadera K, et al. UFT and its metabolites inhibit the angiogenesis induced by murine renal cell carcinoma, as determined by a dorsal air sac assay in mice. Clin Cancer Res. 1999;5:2185–2191. 10473104

35. Colleoni M, Rocca A, Sandri MT, Zorzino L, Masci G, Nolè F, et al. Low-dose oral methotrexate and cyclophosphamide in metastatic breast cancer: antitumor activity and correlation with vascular endothelial growth factor levels. Ann Oncol. 2002;13:73–80.

36. Allegrini G, Di Desidero T, Barletta MT, Fioravanti A, Orlandi P, Canu B, et al. Clinical, pharmacokinetic and pharmacodynamic evaluations of metronomic UFT and cyclophosphamide plus celecoxib in patients with advanced refractory gastrointestinal cancers. Angiogenesis. 2012;15:275–286. doi: 10.1007/s10456-012-9260-6 22382585

37. Nagai N, Mukai K, Hirata E, Jin HH, Komatsu M, Yunokawa M. UFT and its metabolite gamma-butyrolactone (GBL) inhibit angiogenesis induced by vascular endothelial growth factor in advanced cervical carcinoma. Med Oncol. 2008;25:214–221. doi: 10.1007/s12032-007-9023-1 17968681

38. Ueda H, Tanaka H, Kida Y, Fukuchi H, Ichinose M. Adjuvant chemotherapy with tegafur/uracil administration after transcatheter arterial chemoembolization for advanced hepatocellular carcinoma. Oncol Rep. 2008;19:1355–1361. 18425398

39. Ebos JM, Lee CR, Cruz-Munoz W, Bjarnason GA, Christensen JG, Kerbel RS. Accelerated metastasis after short-term treatment with a potent inhibitor of tumour angiogenesis. Cancer Cell. 2009;15:232–239. doi: 10.1016/j.ccr.2009.01.021 19249681

40. Tanaka F. UFT (tegafur and uracil) as postoperative adjuvant chemotherapy for solid tumours (carcinoma of the lung, stomach, colon/rectum, and breast): clinical evidence, mechanism of action, and future direction. Surg Today. 2007;37:923–943. doi: 10.1007/s00595-007-3578-5 17952521

41. Penel N, Adenis A, Bocci G. Cyclophosphamide-based metronomic chemotherapy: after 10 years of experience, where do we stand and where are we going? Crit Rev Oncol Hematol. 2012;82:40–50. doi: 10.1016/j.critrevonc.2011.04.009 21641231

42. Saunders NA, Simpson F, Thompson EW, Hill MM, Endo-Munoz L, Leggatt G, et al. Role of intratumoural heterogeneity in cancer drug resistance: molecular and clinical perspectives. EMBO Mol Med. 2012;4:675–684. doi: 10.1002/emmm.201101131 22733553

43. Harris AL. Hypoxia-a key regulatory factor in tumour growth. Nat Rev Cancer. 2002;2:38–47. doi: 10.1038/nrc704 11902584

44. Patten SG, Adamcic U, Lacombe K, Minhas K, Skowronski K, Coomber BL. VEGFR2 heterogeneity and response to anti-angiogenic low dose metronomic cyclophosphamide treatment. BMC Cancer. 2010 Dec 15;10:683. doi: 10.1186/1471-2407-10-683 21159176

45. Daenen LG, Shaked Y, Man S, Xu P, Voest EE, Hoffman RR, et al. Low-dose metronomic cyclophosphamide combined with vascular disrupting therapy induces potent antitumor activity in preclinical human tumor xenografts models. Mol Cancer Ther. 2009;8:2872–2881. doi: 10.1158/1535-7163.MCT-09-0583 19825805

46. Chen MH, Yip GW, Tse GM, Moriya T, Lui PC, Zin ML, et al. Expression of basal keratins and vimentin in breast cancers of young women correlates with adverse pathologic parameters. Mod Pathol. 2008;21:1183–1191. doi: 10.1038/modpathol.2008.90 18536655

47. Xiong G, Deng L, Zhu J, Rychahou PG, Xu R. Prolyl-4-hydroxylase α subunit 2 promotes breast cancer progression and metastasis by regulating collagen deposition. BMC Cancer. 2014 Jan 2;14:1. doi: 10.1186/1471-2407-14-1 24383403

48. Cheang MCU, Voduc D, Bajdik C, Leung S, MacKinney S, Chia SK, et al. Basal-like breast cancer defined by five biomarkers has superior prognostic value than triple-negative phenotype. Clin Cancer Res. 2008;14:1368–1376. doi: 10.1158/1078-0432.CCR-07-1658 18316557

49. Rakha EA, Elsheikh SE, Aleskandarany MA, Habashi HO, Green AR, Powe DG, et al. Triple-negative breast cancer: distinguishing between basal and non-basal subtypes. Clin Cancer Res. 2009; 15:2302–2310. doi: 10.1158/1078-0432.CCR-08-2132 19318481

50. Charaffe-Jauffret E, Ginestier C, Monville F, Finetti P, Adélaïde J, Cervera N, et al. Gene expression profiling of breast cell lines identifies potential new basal markers. Oncogene. 2006;25:2273–2284. doi: 10.1038/sj.onc.1209254 16288205

51. Knight JF, Lesurf R, Zhao H, Pinnaduwage D, Davis RR, Saleh SM, et al. Met synergizes with p53 loss to induce mammary tumors that possess features of claudin-low breast cancer. Proc Natl Acad Sci U S A. 2013;110:E1301–1310. doi: 10.1073/pnas.1210353110 23509284

52. Kang JY, Dolled-Filhart M, Ocal IT, Singh B, Lin CY, Dickson RB, et al. Tissue microarray analysis of hepatocyte growth factor/Met pathway components reveals a role for Met, matriptase, and hepatocyte growth factor activator inhibitor 1 in the progression of node-negative breast cancer. Cancer Res. 2003; 63:1101–1105. 12615728

53. Xie Y, Lu W, Liu S, Yang Q, Carver BS, Li E, et al. Crosstalk between nuclear MET and SOX9/β-catenin correlates with castration-resistant prostate cancer. Mol Endocrinol. 2014;28:1629–1639. doi: 10.1210/me.2014-1078 25099011

54. Pozner-Moulis S, Pappas DJ, Rimm DL. Met, the hepatocyte growth factor receptor, localizes to the nucleus in cells at low density. Cancer Res. 2006;66:7976–7982. doi: 10.1158/0008-5472.CAN-05-4335 16912172

55. Lovitt CJ, Shelper TB, Avery VM. Evaluation of chemotherapeutics in a three-dimensional breast cancer model. J Cancer Res Clin Oncol. 2015;141:951–959. doi: 10.1007/s00432-015-1950-1 25773123

56. Vidi P-A, Bissell MJ, Lelièvre SA. Three-dimensional culture of human breast epithelial cells: the how and the way. Methods Mol Biol. 2013;945:193–219. doi: 10.1007/978-1-62703-125-7_13 23097109

57. Kenny PA, Lee GY, Myers CA, Neve RM, Semeiks JR, Spellman PT, et al. The morphologies of breast cancer cell lines in three-dimensional assays correlate with their profiles of gene expression. Mol Oncol. 2007;1:84–96. doi: 10.1016/j.molonc.2007.02.004 18516279

58. Friedl P, Locker J, Sahai E, Segall JE. Classifying collective cancer cell invasion. Nat Cell Biol. 2012;14:777–783. doi: 10.1038/ncb2548 22854810

59. Viloria-Petit AM, David L, Jia JY, Erdemir T, Bane AL, Pinnaduwage D, et al. A role for the TGFbeta-Par6 polarity pathway in breast cancer progression. Proc Natl Acad Sci U S A. 2009;106: 14028–14033. doi: 10.1073/pnas.0906796106 19667198

60. Manni A, Washington S, Griffith JW, Verderame MF, Mauger D, Demers LM, et al. Influence of polyamines on in vitro and in vivo features of aggressive and metastatic behavior by human breast cancer cells. Clin Exp Metastasis. 2002;19:95–105. 11964084

61. Mai A, Muharram G, Barrow-McGee R, Baghirov H, Rantala J, Kermorgant S, et al. Distinct c-Met activation mechanisms induce cell rounding or invasion through pathways involving integrins, RhoA and HIP1. J Cell Sci. 2014;127:1938–1952. doi: 10.1242/jcs.140657 24790222

62. Fujii M, Takayama T, Kochi M. Clinical identification of colorectal cáncer patients benefiting from adjuvant uracil-tegafur (UFT): a randomized controlled trial. J Cancer Res Clin Oncol. 2008;134:1319–1323. doi: 10.1007/s00432-008-0417-z 18512074

63. Black PC, Shetty A, Brown GA, Esparza-Coss E, Metwalli AR, Agarwal PK, et al. Validating bladder cancer xenograft bioluminescence with magnetic resonance imaging: the significance of hypoxia and necrosis. BJUI Int. 2010;106:1799–1804.

64. Ponzo MG, Park M. The Met receptor tyrosine kinase and basal breast cancer. Cell cycle. 2010;9:1043–1050. doi: 10.4161/cc.9.6.11033 20237428

65. Gaule PB, Crown J, O´Donovan N, Duffy MJ. cMET in triple-negative breast cancer: is it a therapeutic target for this subset of breast cancer patients? Expert Opin Ther Targets. 2014;18:999–1009. doi: 10.1517/14728222.2014.938050 25084805

66. Inoue T, Kataoka H, Goto K, Nagaike K, Igami K, Naka D, et al. Activation of c-Met (hepatocyte growth factor receptor) in human gastric cancer tissue. Cancer Sci. 2004;95:803–808. doi: 10.1111/j.1349-7006.2004.tb02185.x 15504247

67. Puri N, Ahmed S, Janamanchi V, Tretiakova M, Zumba O, Krausz T, et al. c-Met is a potentially new therapeutic target for treatment of human melanoma. Clin Cancer Res. 2007;13:2246–2253. doi: 10.1158/1078-0432.CCR-06-0776 17404109

68. Ho-Yen CM, Jones JL, Kermogant S. The clinical and functional significance of c-Met in breast cancer: a review. Breast Cancer Res. 2015;17(52). doi: 10.1186/s13058-015-0547-6 25887320

69. Zucali PA, Ruiz MG, Giovannetti E, Destro A, Varella-Garcia M, Floor K, et al. Role of c-MET expression in non-small-cell lung cancer patients treated with EGFR tyrosine kinase inhibitors. Ann Oncol. 2008;19:1605–1612. doi: 10.1093/annonc/mdn240 18467317

70. Barrow-McGee R, Kermorgant S. Met endosomal signalling: in the right place, at the right time. Int J Biochem Cell Biol. 2014;49:69–74. doi: 10.1016/j.biocel.2014.01.009 24440758

71. Kermorgant S, Parker PJ. c-Met signaling: spatio-temporal decisions. Cell Cycle. 2005;4:352–355. doi: 10.4161/cc.4.3.1519 15701970

72. MacLean KE, Vickaryous MK. A novel amniote model of epimorphic regeneration: the leopard gecko, Eublepharis macularius. BMC Dev Biol. 2011;11(50). Available from: https://doi.org/10.1186/1471-213X-11-50

73. Hirsch FR, Varella-Garcia M, Bunn PA Jr, Di Maria MV, Veve R, Bremmes RM, et al. Epidermal growth factor receptor in non-small-cell lung carcinomas: correlation between gene copy number and protein expression and impact on prognosis. J Clin Oncol. 2003;21:3798–3807. doi: 10.1200/JCO.2003.11.069 12953099

74. John T, Liu G, Tsao M-S. Overview of molecular testing in non-small-cell lung cancer: Mutational analysis, gene copy number, protein expression and other biomarkers of EGFR for the prediction of response to tyrosine kinase inhibitors. Oncogene. 2009;28:S14–S23. doi: 10.1038/onc.2009.197 19680292

75. Bocci G, Nicolaou KC, Kerbel RS. Protracted low-dose effects on human endothelial cell proliferation and survival in vitro reveal a selective antiangiogenic window for various chemotherapeutic drugs. Cancer Res. 2002;62:6938–6943. 12460910

76. Benton G, Kleinman HK, George J, Arnaoutova I. Multiple uses of basement membrane-like matrix (BME/Matrigel) in vitro and in vivo with cancer cells. Int J Cancer. 2011;128:1751–1757. doi: 10.1002/ijc.25781 21344372

Suppressive impact of metronomic chemotherapy using UFT and/or cyclophosphamide on mediators of breast cancer dissemination and invasion

Souhrn

Klíčová slova:

Zdroje

PLOS One

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