Tumor Biology

, Volume 34, Issue 2, pp 1245–1253 | Cite as

Molecular diagnosis and prognostic significance of lymph node micrometastasis in patients with histologically node-negative non-small cell lung cancer

  • Chun-Hua Dai
  • Jian Li
  • Li-Chao Yu
  • Xia-Qin Li
  • Shun-Bing Shi
  • Jian-Rong Wu
Research Article


Lymph node metastasis is a major prognostic factor in resected non-small cell lung cancer (NSCLC). However, 30–40 % rate of recurrence after performing complete resection in node-negative patients suggests that their nodal staging is suboptimal. We aimed to evaluate the molecular diagnosis and prognostic significance of lymph node micrometastasis in patients with node-negative NSCLC. Primary tumor samples from 62 patients with resected stage I–IIB NSCLC were screened for fragile histidine triad (FHIT) and CDKN2A mRNA deletion using reverse transcriptase polymerase chain reaction (RT-PCR). The molecular alternations were found in tumors of 49 patients. A total of 269 lymph nodes from these 49 NSCLC patients with FHIT or/and CDKN2A deletion tumors were examined. Fifteen positive-control nodes and ten negative-control nodes were also analyzed for FHIT and CDKN2A mRNA deletion. Thirty-nine (22 %) and 22 (18 %) lymph nodes from the 49 patients with FHIT and CDKN2A mRNA deletion in primary tumor had FHIT and CDKN2A mRNA deletion, respectively. The types of FHIT and CDKN2A mRNA deletion in lymph nodes were identical with those in their primary tumors. By combination of two markers, 16 patients (32.7 %) were found to have nodal micrometastasis. Survival analysis showed that patients with nodal micrometastasis had reduced disease-free survival (P = 0.001) and overall survival (P = 0.002) rates. Multivariate analysis demonstrated that nodal micrometastasis was an independent predictor for worse prognosis. Thus, the detection of lymph node micrometastasis by FHIT and CDKN2A mRNA deletion RT-PCR will be helpful to predict the recurrence and prognosis of patients with completely resected stage I–IIB NSCLC.


Non-small cell lung cancer Lymph node Micrometastasis FHIT CDKN2A 


Conflicts of interest



  1. 1.
    Rena O, Oliaro A, Cavallo A, Filosso PL, Donatic G, Di Marzio P, et al. Stage I non-small cell lung carcinoma: really an early stage? Eur J Cardio-thorac Surg. 2002;21:514–9.CrossRefGoogle Scholar
  2. 2.
    Martini N, Bains MS, Burt ME, Zakowsk MF, McCormack P, Rusch VW, et al. Incidence of local recurrence and second primary tumors in resected stage I lung cancer. J Thoracic Cardiovasc Surg. 1995;109:120–9.CrossRefGoogle Scholar
  3. 3.
    Mountain CF. Revision in the international system for staging lung cancer. Chest. 1997;111:1710–7.PubMedCrossRefGoogle Scholar
  4. 4.
    Ohta Y, Oda M, Wu J, Tsunezuka Y, Hiroshi M, Nonomura A, et al. Can tumor size be a guide for limited surgical intervention in patients with peripheral non-small cell lung cancer? Assessment from the point of view of nodal micrometastasis. J Thorac Cardiovasc Surg. 2001;122:900–6.PubMedCrossRefGoogle Scholar
  5. 5.
    Gu CD, Osaki T, Oyama T, Inoue M, Kodate M, Dobashi K, et al. Detection of micrometastatic tumor-cells in pN0 lymph nodes of patients with completely resected non-small cell lung cancer: impact on recurrence and survival. Ann Surg. 2002;235:133–9.PubMedCrossRefGoogle Scholar
  6. 6.
    Wu J, Ohta Y, Minato H, Tsunezuka Y, Oda M, Watanabe Y, et al. Nodal occult metastasis in patients with peripheral lung adenocarcinoma of 2.0 cm or less in diameter. Ann Thorac Surg. 2001;71:1772–7.PubMedCrossRefGoogle Scholar
  7. 7.
    Rena O, Carsana L, Cristina S, Papalia E, Massera F, Errico L, et al. Lymph node isolated tumor cells and micrometastases in pathological stage I non-small cell lung cancer: prognostic significance. Eur J Cardio-thorac Surg. 2007;32:863–7.CrossRefGoogle Scholar
  8. 8.
    Mosotti M, Falleni M, Palleschi A, Pellegrini C, Alessi F, Bosari S, et al. Quantitative real-time polymerase chain reaction detection of lymph node lung cancer micrometastasis using carcinoembryonic antigen marker. Chest. 2005;128:1539–44.CrossRefGoogle Scholar
  9. 9.
    Saintigny P, Coulon S, Kambouchner M, Ricci S, Martinot E, Dancel C, et al. Real-time RT-PCR detection of CK19 CK7 and MUC1 mRNA for diagnosis of lymph node micrometastases in non small cell lung carcinoma. Int J Cancer. 2005;115:777–82.PubMedCrossRefGoogle Scholar
  10. 10.
    Benlloch S, Galbis-Caravajal JM, Alenda C, Peiro FM, Sanche-Ronco M, Rodriguez-Paniagua JM, et al. Expression of molecular markers in mediastinal nodes from rescted stage I non-small-cell lung cancer (NSCLC): prognostic impact and potential role as markers of occult micrometastases. Ann Oncol. 2009;20:91–7.PubMedCrossRefGoogle Scholar
  11. 11.
    Ahrendt SA, Yang SC, Wu L, Roing CM, Russell P, Westra WH, et al. Molecular assessment of lymph nodes in patients with resected stage I non-small cell lung cancer: preliminary results of a prospective study. J Thoracic Cardiovasc Surg. 2002;123:466–73.CrossRefGoogle Scholar
  12. 12.
    Wang XT, Sienel W, Eggeling S, Luduig C, Stoelben E, Mueller J, et al. Detection of disseminated tumor cells in mediastinoscopic lymph node biopsies and lymphadenectomy specimens of patients with NSCLC by quantitative RT-PCR. Eur J Cardio-Thorac Surg. 2005;28:26–32.CrossRefGoogle Scholar
  13. 13.
    Ohta M, Inoue H, Cottielli MG, Kastury K, Baffa R, Palazzo J, et al. The FHIT gene, spanning the chromosome 3P14.2 fragile site and renal carcinoma-associated t(3:8) breakpoint is abnormal in digestive tract cancer. Cell. 1996;84:587–97.PubMedCrossRefGoogle Scholar
  14. 14.
    Crose CM, Sozzi G, Huebner K. Role of FHIT in human cancer. J Clin Oncol. 1999;17:1618–24.Google Scholar
  15. 15.
    Pekarsky Y, Palamarchuk A, Huebner K, Croce CM. FHIT as tumor suppressor: mechanisms and therapeutic opportunities. Cancer Biol Ther. 2002;1:233–6.Google Scholar
  16. 16.
    Wali A, Srinivasan R, Shabnam MS, Majumler S, Joshi K, Behera D. Loss of fragile histidine triad gene expression in advanced lung cancer is consequent to allelic loss at 3P14 locus and promoter methylation. Mol Cancer Res. 2006;4:93–9.PubMedCrossRefGoogle Scholar
  17. 17.
    Verri C, Roz L, Conte D, Liloglou T, Livio A, Vesin A, et al. Fragile histidine triad gene inactivation in lung cancer. The European Early Lung Cancer project. Am J Respir Crit Care Med. 2009;179:396–401.PubMedCrossRefGoogle Scholar
  18. 18.
    Zbchbauer-Muller S, Wistuba II, Minna TD, Gazdar AF. Fragile histidine triad (FHIT) gene abnormalities in lung cancer. Clin Lung Cancer. 2000;2:141–5.CrossRefGoogle Scholar
  19. 19.
    Herman JG, Merlo A, Mao L, Lapidus RG, Issa JP, Davidson NE, et al. Inactivation of the CDKN2/P16/MTS1 gene is frequently associated with aberrant DNA methylation in all common human cancer. Cancer Res. 1995;55:4525–30.PubMedGoogle Scholar
  20. 20.
    Vonlanthen S, Heighway J, Tschan J, Tschan MP, Borner MM, Altermatt HJ, et al. Expression of P16 INK4a/P16 alpha and P19ARF/P16 beta is frequently altered in non-small cell lung cancer and correlates with p53 overexpression. Oncogene. 1998;17:2779–85.PubMedCrossRefGoogle Scholar
  21. 21.
    Belisky SA, Nikula KJ, Palmisano WA, Michels R, Saccomanno G, Gabrielson E, et al. Aberrant methylation of P16INK4a is an early event in lung cancer and a potential biomarker for early diagnosis. Proc Natl Acad Sci U S A. 1998;95:11891–6.CrossRefGoogle Scholar
  22. 22.
    Nobori T, Miura K, Wu DJ, Lois S, Takabayashi K, Carson DA. Deletion of the cyclin-dependent kiase-4 inhibitor gene in multiple human cancers. Nature. 1994;368:753–6.PubMedCrossRefGoogle Scholar
  23. 23.
    Liggett WH, Sidransky D. Role of the p16 tumor suppressor gene in cancer. J Clin Oncol. 1998;16:1197–206.PubMedGoogle Scholar
  24. 24.
    Gazzeris S, Gouyer V, Vourch C, Brambilla C, Brambilla E. Mechanisms of P16INK4A inactivation in non small-cell lung cancers. Oncogenes. 1998;16:497–504.CrossRefGoogle Scholar
  25. 25.
    Jarmalaite S, Kannio A, Anttila S, Lazutka JR, Husgafvel-Pursiainen K. Aberrant P16 promoter methylation in smokers and former smokers with nonsmall cell lung cancer. Int J Cancer. 2003;106:913–8.PubMedCrossRefGoogle Scholar
  26. 26.
    Tanaka R, Wang D, Morishita Y, Inadome Y, Minami Y, Lijima T, et al. Loss of function of P16 gene and prognosis of pulmonary adenocarcinoma. Cancer. 2005;103:608–15.PubMedCrossRefGoogle Scholar
  27. 27.
    Green FL, Page DL, Fleming ID, et al. AJCC Cancer Staging Manual. 6th ed. New York: Springer; 2002.Google Scholar
  28. 28.
    Hunter T. Cooperation between oncogenes. Cell. 1991;64:249–70.PubMedCrossRefGoogle Scholar
  29. 29.
    Minna JD. Genetic events in the pathogenesis of lung cancer. Chest. 1989;96:17–23s.Google Scholar
  30. 30.
    Panani AD, Roussos C. Cytogenetic and molecular aspects of lung cancer. Cancer Lett. 2006;239:1–9.PubMedCrossRefGoogle Scholar
  31. 31.
    Hashimoto T, Kobayashi Y, Ishikawa Y, Tsuchiya S, Okumura S, Nakagawa K, et al. Prognostic value of genetically diagnosed lymph node micrometastasis in non-small cell lung carcinoma cases. Cancer Res. 2000;15:6472–8.Google Scholar
  32. 32.
    Li S-H, Wang Z, Liu X-Y, Liu F-Y. Gene diagnosis and prognostic significance of lymph node micrometastasis after complete resection of histologically node-negative non-small cell lung cancer. World J Surg. 2008;32:1651–6.PubMedCrossRefGoogle Scholar
  33. 33.
    Osaki T, Oyama T, Gu C-D, Yamashita T, So T, Takenoyam M, et al. Prognostic impact of micrometastatic tumor cells in the lymph nodes and bone marrow of patients with completely resected stage I non-small-cell lung cancer. J Clin Oncol. 2002;20:2930–6.PubMedCrossRefGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2013

Authors and Affiliations

  • Chun-Hua Dai
    • 1
  • Jian Li
    • 2
  • Li-Chao Yu
    • 3
  • Xia-Qin Li
    • 1
  • Shun-Bing Shi
    • 3
  • Jian-Rong Wu
    • 4
  1. 1.Department of Medical OncologyAffiliated Hospital of Jiangsu UniversityZhenjiangChina
  2. 2.Department of Pulmonary MedicineAffiliated Hospital of Jiangsu UniversityZhenjiangChina
  3. 3.Department of Thoracic SurgeryAffiliated Hospital of Jiangsu UniversityZhenjiangChina
  4. 4.Department of PathologyAffiliated Hospital of Jiangsu UniversityZhenjiangChina

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