Abstract
Angiogenesis and lymphangiogenesis are key components of non-small cell lung cancer (NSCLC) tumor growth and metastatic spread; however, the prognostic and predictive role of angiogenic and lymphangiogenic biomarkers remains controversial for NSCLC patients. We assessed VEGF, VEGFC, VEGFD, VEGFR3 protein expression, tumor microvessel, and lymphatic vessel (LmVD) density by immunohistochemistry in 103 NSCLC; biomarkers were analyzed individually as well as multiplexed with each other. No correlations were identified between VEGF, VEGFC, VEGFD, or LmVD and clinical characteristics. VEGFR3 was correlated with VEGFC (p = 0.03), VEGFD (p < 0.0001), and intratumor LmVD (p = 0.03). Tumors that did not express VEGFR3 had a worse prognosis (log rank p = 0.03). VEGF was significantly correlated with survival in adenocarcinomas (log rank p = 0.014) but not in squamous cell carcinomas (log rank p = 0.5). Multivariate Cox regression analysis confirmed the independent prognostic potential of VEGFR3 (hazard ratio (HR) = 0.05; 95% confidence intervals (CI) = 0.008–0.32, p = 0.002) for all patients and VEGF (HR = 8.69, 95% CI = 1.4–53.69, p = 0.02) for adenocarcinomas. When biomarkers were multiplexed, only stage and VEGFC expression were independent predictors of survival for all patients. Weighted expression of VEGFC, VEGFR3, and stage was used to build a prognostic classifier for stage I–IIIA patients; patients in the low risk group had prolonged survival compared with high risk patients (log rank p = 0.02). There was no association between biomarkers and early recurrence or response to treatment. Angiogenic and lymphangiogenic biomarkers studied define subgroups of patients at high risk and may be useful for prognostic stratification of NSCLC patients especially those with early stage disease.
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References
Bergers G, Benjamin LE (2003) Tumorigenesis and the angiogenic switch. Nat Rev Cancer 3(6):401–410. doi:10.1038/nrc1093
Gasparini G, Longo R, Toi M, Ferrara N (2005) Angiogenic inhibitors: a new therapeutic strategy in oncology. Nat Clin Pract Oncol 2(11):562–577. doi:10.1038/ncponc0342
Ferrara N, Gerber HP, LeCouter J (2003) The biology of vegf and its receptors. Nat Med 9(6):669–676. doi:10.1038/nm0603-669
Sandler A, Gray R, Perry MC, Brahmer J, Schiller JH, Dowlati A, Lilenbaum R, Johnson DH (2006) Paclitaxel-carboplatin alone or with bevacizumab for non-small-cell lung cancer. N Engl J Med 355(24):2542–2550. doi:10.1056/NEJMoa061884
El-Chemaly S, Levine SJ, Moss J (2008) Lymphatics in lung disease. Ann NY Acad Sci 1131:195–202. doi:10.1196/annals.1413.017
Achen MG, Stacker SA (2008) Molecular control of lymphatic metastasis. Ann NY Acad Sci 1131:225–234. doi:10.1196/annals.1413.020
Stacker SA, Achen MG, Jussila L, Baldwin ME, Alitalo K (2002) Lymphangiogenesis and cancer metastasis. Nat Rev Cancer 2(8):573–583. doi:10.1038/nrc863
Renyi-Vamos F, Tovari J, Fillinger J, Timar J, Paku S, Kenessey I, Ostoros G, Agocs L, Soltesz I, Dome B (2005) Lymphangiogenesis correlates with lymph node metastasis, prognosis, and angiogenic phenotype in human non-small cell lung cancer. Clin Cancer Res 11(20):7344–7353
Kadota K, Huang CL, Liu D, Ueno M, Kushida Y, Haba R, Yokomise H (2008) The clinical significance of lymphangiogenesis and angiogenesis in non-small cell lung cancer patients. Eur J Cancer 44(7):1057–1067
Zhan P, Wang J, Lv XJ, Wang Q, Qiu LX, Lin XQ, Yu LK, Song Y (2009) Prognostic value of vascular endothelial growth factor expression in patients with lung cancer: a systematic review with meta-analysis. J Thorac Oncol 4(9):1094–1103. doi:10.1097/JTO.0b013e3181a97e31
de Santa C, Pau E, Arias FC, Caso Pelaez E, Munoz Molina GM, Sanchez Hernandez I, Muguruza Trueba I, Moreno Balsalobre R, Sacristan Lopez S, Gomez Pinillos A, del Val Toledo Lobo M (2009) Prognostic significance of the expression of vascular endothelial growth factors a, b, c, and d and their receptors r1, r2, and r3 in patients with nonsmall cell lung cancer. Cancer 115(8):1701–1712
Travis WD, Brambilla E, Muller-Hermelink HK, Harris CC (2004) World health organization classification of tumors. Pathology and genetics of tumors of the lung, pleura, thymus and heart. IARC Press, Lyon
Therasse P, Arbuck SG, Eisenhauer EA, Wanders J, Kaplan RS, Rubinstein L, Verweij J, Van Glabbeke M, van Oosterom AT, Christian MC, Gwyther SG (2000) New guidelines to evaluate the response to treatment in solid tumors. European organization for research and treatment of cancer, national cancer institute of the united states, national cancer institute of canada. J Natl Cancer Inst 92(3):205–216
Adachi Y, Nakamura H, Kitamura Y, Taniguchi Y, Araki K, Shomori K, Horie Y, Kurozawa Y, Ito H, Hayashi K (2007) Lymphatic vessel density in pulmonary adenocarcinoma immunohistochemically evaluated with anti-podoplanin or anti-d2-40 antibody is correlated with lymphatic invasion or lymph node metastases. Pathol Int 57(4):171–177
Andersen S, Donnem T, Al-Saad S, Al-Shibli K, Busund LT, Bremnes RM (2009) Angiogenic markers show high prognostic impact on survival in marginally operable non-small cell lung cancer patients treated with adjuvant radiotherapy. J Thorac Oncol 4(4):463–471. doi:10.1097/JTO.0b013e3181991d18
Arinaga M, Noguchi T, Takeno S, Chujo M, Miura T, Uchida Y (2003) Clinical significance of vascular endothelial growth factor c and vascular endothelial growth factor receptor 3 in patients with nonsmall cell lung carcinoma. Cancer 97(2):457–464. doi:10.1002/cncr.11073
Bo C, Xiaopeng D, Chuanliang P, Xiaogang Z (2009) Expression of vascular endothelial growth factors c and d correlates with lymphangiogenesis and lymph node metastasis in lung adenocarcinoma. Thorac Cardiovasc Surg 57(5):291–294. doi:10.1055/s-0029-1185625
Saintigny P, Kambouchner M, Ly M, Gomes N, Sainte-Catherine O, Vassy R, Czernichow S, Letoumelin P, Breau JL, Bernaudin JF, Kraemer M (2007) Vascular endothelial growth factor-c and its receptor vegfr-3 in non-small-cell lung cancer: concurrent expression in cancer cells from primary tumour and metastatic lymph node. Lung cancer (Amsterdam, Netherlands) 58(2):205–213
Maekawa S, Iwasaki A, Shirakusa T, Enatsu S, Kawakami T, Kuroki M, Kuroki M (2007) Correlation between lymph node metastasis and the expression of vegf-c, vegf-d and vegfr-3 in t1 lung adenocarcinoma. Anticancer Res 27(6A):3735–3741
Kojima H, Shijubo N, Yamada G, Ichimiya S, Abe S, Satoh M, Sato N (2005) Clinical significance of vascular endothelial growth factor-c and vascular endothelial growth factor receptor 3 in patients with t1 lung adenocarcinoma. Cancer 104(8):1668–1677
Chen F, Takenaka K, Ogawa E, Yanagihara K, Otake Y, Wada H, Tanaka F (2004) Flt-4-positive endothelial cell density and its clinical significance in non-small cell lung cancer. Clin Cancer Res 10(24):8548–8553
Trivella M, Pezzella F, Pastorino U, Harris AL, Altman DG (2007) Microvessel density as a prognostic factor in non-small-cell lung carcinoma: a meta-analysis of individual patient data. Lancet Oncol 8(6):488–499. doi:10.1016/S1470-2045(07)70145-6
Sun JG, Wang Y, Chen ZT, Zhuo WL, Zhu B, Liao RX, Zhang SX (2009) Detection of lymphangiogenesis in non-small cell lung cancer and its prognostic value. J Exp Clin Cancer Res 28:21
Hoffman PC, Mauer AM, Vokes EE (2000) Lung cancer. Lancet 355(9202):479–485. doi:10.1016/S0140-6736(00)82038-3
Weir BA, Woo MS, Getz G, Perner S, Ding L, Beroukhim R, Lin WM, Province MA, Kraja A, Johnson LA, Shah K, Sato M, Thomas RK, Barletta JA, Borecki IB, Broderick S, Chang AC, Chiang DY, Chirieac LR, Cho J, Fujii Y, Gazdar AF, Giordano T, Greulich H, Hanna M, Johnson BE, Kris MG, Lash A, Lin L, Lindeman N, Mardis ER, McPherson JD, Minna JD, Morgan MB, Nadel M, Orringer MB, Osborne JR, Ozenberger B, Ramos AH, Robinson J, Roth JA, Rusch V, Sasaki H, Shepherd F, Sougnez C, Spitz MR, Tsao MS, Twomey D, Verhaak RG, Weinstock GM, Wheeler DA, Winckler W, Yoshizawa A, Yu S, Zakowski MF, Zhang Q, Beer DG, Wistuba II, Watson MA, Garraway LA, Ladanyi M, Travis WD, Pao W, Rubin MA, Gabriel SB, Gibbs RA, Varmus HE, Wilson RK, Lander ES, Meyerson M (2007) Characterizing the cancer genome in lung adenocarcinoma. Nature 450(7171):893–898. doi:10.1038/nature06358
Yanagisawa K, Shyr Y, Xu BJ, Massion PP, Larsen PH, White BC, Roberts JR, Edgerton M, Gonzalez A, Nadaf S, Moore JH, Caprioli RM, Carbone DP (2003) Proteomic patterns of tumour subsets in non-small-cell lung cancer. Lancet 362(9382):433–439. doi:10.1016/S0140-6736(03)14068-8
Donnem T, Al-Shibli K, Al-Saad S, Delghandi MP, Busund LT, Bremnes RM (2009) Vegf-a and vegfr-3 correlate with nodal status in operable non-small cell lung cancer: Inverse correlation between expression in tumor and stromal cells. Lung cancer (Amsterdam, Netherlands) 63(2):277–283
Ko YH, Jung CK, Lee MA, Byun JH, Kang JH, Lee KY, Jo KH, Wang YP, Hong YS (2008) Clinical significance of vascular endothelial growth factors (vegf)-c and -d in resected non-small cell lung cancer. Cancer Res Treat 40(3):133–140
Takizawa H, Kondo K, Fujino H, Kenzaki K, Miyoshi T, Sakiyama S, Tangoku A (2006) The balance of vegf-c and vegfr-3 mrna is a predictor of lymph node metastasis in non-small cell lung cancer. Br J Cancer 95(1):75–79
Takanami I (2006) Lymphatic microvessel density using d2-40 is associated with nodal metastasis in non-small cell lung cancer. Oncol Rep 15(2):437–442
Kopfstein L, Veikkola T, Djonov VG, Baeriswyl V, Schomber T, Strittmatter K, Stacker SA, Achen MG, Alitalo K, Christofori G (2007) Distinct roles of vascular endothelial growth factor-d in lymphangiogenesis and metastasis. Am J Pathol 170(4):1348–1361
He Y, Rajantie I, Pajusola K, Jeltsch M, Holopainen T, Yla-Herttuala S, Harding T, Jooss K, Takahashi T, Alitalo K (2005) Vascular endothelial cell growth factor receptor 3-mediated activation of lymphatic endothelium is crucial for tumor cell entry and spread via lymphatic vessels. Cancer Res 65(11):4739–4746. doi:10.1158/0008-5472.CAN-04-4576
Dixelius J, Makinen T, Wirzenius M, Karkkainen MJ, Wernstedt C, Alitalo K, Claesson-Welsh L (2003) Ligand-induced vascular endothelial growth factor receptor-3 (vegfr-3) heterodimerization with vegfr-2 in primary lymphatic endothelial cells regulates tyrosine phosphorylation sites. J Biol Chem 278(42):40973–40979. doi:10.1074/jbc.M304499200
Naumnik W, Izycki T, Swidzinska E, Ossoliniska M, Chyczewska E (2007) Serum levels of vegf-c, vegf-d, and svegf-r2 in patients with lung cancer during chemotherapy. Oncol Res 16(9):445–451
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Anagnostou, V.K., Tiniakos, D.G., Fotinou, M. et al. Multiplexed analysis of angiogenesis and lymphangiogenesis factors predicts outcome for non-small cell lung cancer patients. Virchows Arch 458, 331–340 (2011). https://doi.org/10.1007/s00428-010-1015-4
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DOI: https://doi.org/10.1007/s00428-010-1015-4