Circulating Tumor Cells Identify Early Recurrence in Patients with Non-Small Cell Lung Cancer Undergoing Radical Resection

Clara Bayarri-Lara, Francisco G Ortega, Antonio Cueto Ladrón de Guevara, Jose L Puche, Javier Ruiz Zafra, Diego de Miguel-Pérez, Abel Sánchez-Palencia Ramos, Carlos Fernando Giraldo-Ospina, Juan A Navajas Gómez, Miguel Delgado-Rodriguez, Jose A Lorente, María Jose Serrano, Clara Bayarri-Lara, Francisco G Ortega, Antonio Cueto Ladrón de Guevara, Jose L Puche, Javier Ruiz Zafra, Diego de Miguel-Pérez, Abel Sánchez-Palencia Ramos, Carlos Fernando Giraldo-Ospina, Juan A Navajas Gómez, Miguel Delgado-Rodriguez, Jose A Lorente, María Jose Serrano

Abstract

Background: Surgery is the treatment of choice for patients with non-small cell lung cancer (NSCLC) stages I-IIIA. However, more than 20% of these patients develop recurrence and die due to their disease. The release of tumor cells into peripheral blood (CTCs) is one of the main causes of recurrence of cancer. The objectives of this study are to identify the prognostic value of the presence and characterization of CTCs in peripheral blood in patients undergoing radical resection for NSCLC.

Patients and methods: 56 patients who underwent radical surgery for previously untreated NSCLC were enrolled in this prospective study. Peripheral blood samples for CTC analysis were obtained before and one month after surgery. In addition CTCs were phenotypically characterized by epidermal growth factor receptor (EGFR) expression.

Results: 51.8% of the patients evaluated were positive with the presence of CTCs at baseline. A decrease in the detection rate of CTCs was observed in these patients one month after surgery (32.1%) (p = 0.035). The mean number of CTCs was 3.16 per 10 ml (range 0-84) preoperatively and 0.66 (range 0-3) in postoperative determination. EGFR expression was found in 89.7% of the patients at baseline and in 38.9% patients one month after surgery. The presence of CTCs after surgery was significantly associated with early recurrence (p = 0.018) and a shorter disease free survival (DFS) (p = .008). In multivariate analysis CTC presence after surgery (HR = 5.750, 95% CI: 1.50-21.946, p = 0.010) and N status (HR = 0.296, 95% CI: 0.091-0.961, p = 0.043) were independent prognostic factors for DFS.

Conclusion: CTCs can be detected and characterized in patients undergoing radical resection for non-small cell lung cancer. Their presence might be used to identify patients with increased risk of early recurrence.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Fig 1. Image galleries after isolation, cytomorphological…
Fig 1. Image galleries after isolation, cytomorphological analysis, and detection of cytokeratin-positive (CK+) cells (red staining) and epidermal growth factor receptor (EGFR).
A) T1975 cell tumor line. was used as positive control for EGFR expression B) A control in which the slides are incubated with antibody diluent, without the primary antibody was included. (C, D) Expression of different markers in patients with non-small lung cancer through combination of stained CK+ cells (red) with EGFR (blue). EGFR-specific immunofluorescence (IF) of circulating tumor cells (CTCs) was determined with Alexa 355.
Fig 2
Fig 2
Kaplan-Meier curves of disease free survival according to CTC status before (A) and one month after surgery (B). CTC1: CTC presence before surgery. CTC2: CTC presence after surgery.

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