Observation of circulating tumour cells in patients with non-small cell lung cancer by real-time fluorescent quantitative reverse transcriptase-polymerase chain reaction in peroperative period

  • Ming Jian Ge
  • De Shi
  • Qing Chen Wu
  • Mei Wang
  • Liang Bin Li
Original Paper


Purpose: To assess whether surgical manoeuvre or resection of lung cancer could lead to haematogenous dissemination of malignant cells. In the mean time, the relationship between the sequence of vessel ligation and the haematogenous dissemination of cancer cells during operation was determined. Methods: Exploiting cytokeratin 19 (CK19)/carcinoembryonic antigen (CEA) mRNA as markers, 69 peripheral blood samples were collected from 23 consecutive patients with non-small cell lung cancer (NSCLC) who underwent surgical resection with curative intention in preoperative, intraoperative and postoperative period, respectively. Before the operation, all patients were randomly assigned to one of the two surgical procedure groups according to the order of vessel ligation, PV-first group and PA-first group. Additionally, the ten patients with benign lung disease served as control subjects undergoing surgical resection. The quantity and timing of the shedding of lung cancer cells into the circulation of patients were also monitored by fluorescent quantitative-reverse transcriptase-polymerase chain reaction before, during and after surgery. Results: (1) The CK19 diagnostic test: the value of CK19 mRNA in operation was significantly higher than that of preoperation (5.246±0.196 vs. 4.472±0.164, P=0.000) and postoperation (5.246±0.196 vs. 4.694±0.177, P=0.013). The values between adenocarcinoma and squamous carcinoma were strikingly different (4.9110±1.0315 vs. 4.1891±0.4126, t=2.364, P=0.028). The values between PV-first group and PA-first group during perioperative period appear to be different (4.503 vs. 5.085, P=0.086). Before operation, of the 23 cases studied, 14 cases were positive (60.9%). Surprisingly, circulating epithelial cells were detected in two patients resected for benign lung disease. (2) The CEA diagnostic test: the level of CEA mRNA ascended continuously within this period. The postoperative values were significantly higher than those of preoperation (4.874 vs. 4.483, P=0.000) and those of operative day (4.874 vs. 4.537, P=0.000). The values between PV-first group and PA-first group appear to reach statistical significance (4.397 vs. 4.817, P=0.075). At the same time, there was a correlation between preoperative T-stage and perioperative CEA mRNA (4.267 vs. 4.760, P=0.025). Among the 23 cases, 10 cases were positive (43.5%). Both patients with benign lung disease served as control subjects undergoing surgical resection and the volunteers were negative. Conclusions: A considerable proportion of patients who appear to have resectable NSCLC might be regarded as having systemic disease, which is often undetectable by current tumour staging method. In terms of a marker used for the NSCLC patients who undergo operation, CEA is more suitable than CK19. The CK19-expressing epithelial cells are released intraoperatively into the circulation, meanwhile CEA-expressing tumour cells are disseminated mostly postoperatively. Surgical manipulation could promote the release of tumour cells into the bloodstream, but the ligation of pulmonary vein before the ligation of the pulmonary artery may partly prevent such release during surgery.


Lung neoplasm Blood Polymerase chain reaction Cytokeratin Carcinoembryonic antigen Messenger RNA Peroperative period 



Fluorescent quantitative-reverse transcriptase-polymerase chain reaction


Cytokeratin 19


Carcinoembryonic antigen


Non-small cell lung cancer


Pulmonary vein


Pulmonary artery


Peripheral blood




Squamous cell carcinoma


Peripheral blood mononuclear cells


Curve, receiver operator characteristic curve


Circulating tumour cells


Confidence interval


Threshold cycle


Constant of variance


Minimal residual disease


Tumour-node-metastases staging



I extremely thank Dr. Gaynor Bates, who works in Breast Cancer Campaign in the UK, for her suggestion concerning the revision of the initial English draft of the study. I am indebted to all the members of the Chongqing Lung Cancer Center who have cheerfully donated the samples.


  1. Bockmann B, Grill HJ, Giesing M (2001) Molecular characterization of minimal residual cancer cells in patients with solid tumors. Biomol Eng 17(3):95–111PubMedCrossRefGoogle Scholar
  2. Castaldo G, Tomaiuolo R, Sanduzzi A, Bocchino ML, Ponticiello A, Barra E, Vitale D, Bariffi F, Sacchetti L, Salvatore F (1997) Lung cancer metastatic cells detected in blood by reverse transcriptase-polymerase chain reaction and dot-blot analysis. J Clin Oncol 15(11):3388–3393PubMedGoogle Scholar
  3. Curry BJ, Myers K, Hersey P (1999) MART-1 is expressed less frequently on circulating melanoma cells in patients who develop distant compared with locoregional metastases. J Clin Oncol 17(8):2562–2571PubMedGoogle Scholar
  4. Fidler IJ (1991) The biology of human cancer metastasis. Acta Oncol 30(6):668–675PubMedGoogle Scholar
  5. Ge MJ, Wu QC,Wang M, Zhang YH, Li LB (2005) Detection of disseminated lung cancer cells in regional lymph nodes by assay of CK(19) reverse transcriptase polymerase chain reaction and its clinical significance. J Cancer Res Clin Oncol 131(10):662–668PubMedCrossRefGoogle Scholar
  6. Giesing M, Austrup F, Bockmann B, Driesel G, Eder C, Kusiak I, Suchy B, Uciechowski P, Grill HJ (2000) Independent prognostication and therapy monitoring of breast cancer patients by DNA/RNA typing of minimal residual cancer cells. Int J Biol Markers 15(1):94–99PubMedGoogle Scholar
  7. Hansen E, Wolff N, Knuechel R, Ruschoff J, Hofstaedter F, Taeger K (1995) Tumor cells in blood shed from the surgical field. Arch Surg 130(4):387–393PubMedGoogle Scholar
  8. Hardingham JE, Hewett PJ, Sage RE, Finch JL, Nuttall JD, Kotasek D, Dobrovic A (2000) Molecular detection of blood-borne epithelial cells in colorectal cancer patients and in patients with benign bowel disease. Int J Cancer 89(1):8–13PubMedCrossRefGoogle Scholar
  9. Hosch S, Kraus J, Scheunemann P, Izbicki JR, Schneider C, Schumacher U, Witter K, Speicher MR, Pantel K (2000) Malignant potential and cytogenetic characteristics of occult disseminated tumor cells in esophageal cancer. Cancer Res 60(24):6836–6840PubMedGoogle Scholar
  10. Hughes TP, Morgan GJ, Martiat P, Goldman JM (1991) Detection of residual leukemia after bone marrow transplant for chronic myeloid leukemia: role of polymerase chain reaction in predicting relapse. Blood 77(4):874–878PubMedGoogle Scholar
  11. Kurusu Y, Yamashita J, Ogawa M (1999) Detection of circulating tumor cells by reverse transcriptase-polymerase chain reaction in patients with resectable non-small-cell lung cancer. Surgery 126(5):820–826PubMedGoogle Scholar
  12. Moll R (1994) Cytokeratins in the histological diagnosis of malignant tumors. Int J Biol Markers 9(2):63–69PubMedGoogle Scholar
  13. Offner S, Schmaus W, Witter K, Baretton GB, Schlimok G, Passlick B, Riethmuller G, Pantel K (1999) p53 gene mutations are not required for early dissemination of cancer cells. Proc Natl Acad Sci USA 96(12):6942–6946PubMedCrossRefGoogle Scholar
  14. Pachmann K, Clement JH, Schneider CP, Willen B, Camara O, Pachmann U, Hoffken K (2005) Standardized quantification of circulating peripheral tumor cells from lung and breast cancer. Clin Chem Lab Med 43(6):617–627PubMedCrossRefGoogle Scholar
  15. Pantel K, Cote RJ, Fodstad O (1999) Detection and clinical importance of micrometastatic disease. J Natl Cancer Inst 91(13):1113–1124PubMedCrossRefGoogle Scholar
  16. Peck K, Sher YP, Shih JY, Roffler SR, Wu CW, Yang PC (1998) Detection and quantitation of circulating cancer cells in the peripheral blood of lung cancer patients. Cancer Res 58(13):2761–2765PubMedGoogle Scholar
  17. Rolle A, Gunzel R, Pachmann U, Willen B, Hoffken K, Pachmann K (2005) Increase in number of circulating disseminated epithelial cells after surgery for non-small cell lung cancer monitored by MAINTRAC(R) is a predictor for relapse : a preliminary report. World J Surg Oncol 3(1):18PubMedCrossRefGoogle Scholar
  18. Weitz J, Kienle P, Lacroix J, Willeke F, Benner A, Lehnert T, Herfarth C, von Knebel Doeberitz M (1998) Dissemination of tumor cells in patients undergoing surgery for colorectal cancer. Clin Cancer Res 4(2):343–348PubMedGoogle Scholar
  19. Wong IH, Lau WY, Leung T, Yeo W, Johnson PJ (1999) Hematogenous dissemination of hepatocytes and tumor cells after surgical resection of hepatocellular carcinoma: a quantitative analysis. Clin Cancer Res 5(12):4021–4027PubMedGoogle Scholar
  20. Yamashita JI, Kurusu Y, Fujino N, Saisyoji T, Ogawa M (2000) Detection of circulating tumor cells in patients with non-small cell lung cancer undergoing lobectomy by video-assisted thoracic surgery: a potential hazard for intraoperative hematogenous tumor cell dissemination. J Thorac Cardiovasc Surg 119(5):899–905PubMedCrossRefGoogle Scholar
  21. Zhang JS, Caplin S, Bosman FT, Benhattar J (1997) Genetic diversity at the p53 locus between primary human colorectal adenocarcinomas and their lymph-node metastases. Int J Cancer 70(6):674–678PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Ming Jian Ge
    • 1
  • De Shi
    • 2
  • Qing Chen Wu
    • 1
  • Mei Wang
    • 3
  • Liang Bin Li
    • 1
  1. 1.Department of Thoracic Surgery, The First Affiliated HospitalChongqing University of Medical SciencesChongqingPeople’s Republic of China
  2. 2.Department of General Surgery, The First Affiliated HospitalChongqing University of Medical SciencesChongqingPeople’s Republic of China
  3. 3.Department of Clinical Laboratory, The First Affiliated HospitalChongqing University of Medical SciencesChongqingPeople’s Republic of China

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