Author of

• 25946

  +

  P2.09 - Pathology (Not CME Accredited Session) (ID 958)

  • Event: WCLC 2018
  • Type: Poster Viewing in the Exhibit Hall
  • Track:
  • Presentations: 1
  • Moderators:
  • Coordinates: 9/25/2018, 16:45 - 18:00, Exhibit Hall

  • 27086

    +

    P2.09-28 - Detection of Isolated Tumor Cells in Regional Lymph Nodes from pN0 Lung Cancer by Negative Selection Using Immunomagnetic Beads (ID 12761)

    16:45 - 18:00  |  Author(s): Ganwei Liu
    • Abstract
    • Slides

    Background
    

    Isolated tumor cells (ITC) referred to single tumor cells or small clusters of cells detected by routine hematoxylin and eosin stains or immunohistochemiry. Early detection of ITC in regional lymph nodes could identify a subset of N0 patients with potential risk of disease relapse. This study aimed to establish a new method to detect ITC based on a combination of immunomagnetic bead enrichment, immunofluorescence (IF) and immunofluorescence in situ hybridization (iFISH).

    a9ded1e5ce5d75814730bb4caaf49419 Method
    

    Lymph nodes were obtained from lung cancer patients intraoperatively. After immunomagnetic negative enrichment, tumor cells were identified using IF and iFISH, respectively. P53+/DAPI+/CD45- cells or CEP8+/DAPI+/CD45- cells were judged as positive cells. By comparing pathologically proved metastatic lymph nodes with those from non-invasive adenocarcinoma as control, the best cut-off values of the positive cells number was determinated using the receiver operating characteristic (ROC) curve to predict ITC. Single cell sequencing technology was employed to detect p53+/DAPI+/CD45- cells for TP53 mutation. Pathologically negative lymph nodes were detected for ITC using this newly established method.

    4c3880bb027f159e801041b1021e88e8 Result
    

    Eleven metastatic lymph nodes from ten patients with NSCLC and eighteen lymph nodes from nine patients with non-invasive adenocarcinoma were collected for ROC curve analysis. p53+/DAPI+/CD45- cells were detected in 90.9% metastatic lymph nodes and 44.4% in the control group(P<0.001). The optimal cut-off value for the diagnosis of ITC by IF was 3 positive cells according to ROC curve analysis. The sensitivity and specificity were 81.82% and 94.44%, respectively. Four p53+/DAPI+/CD45- cells were tested using single-cell sequencing technique. All these cells were present with TP53 gene mutations. CEP8+/DAPI+/CD45- cells were detected in 60% metastatic lymph nodes and 6.25% lymph nodes in the control group(P=0.002). The optimal cut-off value for the diagnosis of ITC by iFISH was 1 positive cell. The sensitivity and specificity were 60% and 93.75%, respectively. Combining the two detection techniques, the sensitivity and specificity increased to 90% and 93.75%, respectively. Sixty lymph node samples from thirty patients with invasive NSCLC and pN0 disease were enriched by immunomagnetic beads, detected for ITC by IF, and iFISH. ITC were detected in 18.3%（11/60）of the pN0 lymph node samples. The pN0 status was reclassified as N0(mol+) in 33%(10/30) patients based on the recommendation of the American Joint Committee for Cancer.

    8eea62084ca7e541d918e823422bd82e Conclusion
    

    We established a new method to identify ITC in the regional lymph nodes by a combination of negative enrichment by immunomagnetic bead, IF and iFISH techniques, which could improve the detection of ITC compared with routine pathological examination.

    6f8b794f3246b0c1e1780bb4d4d5dc53

    Only Active Members that have purchased this event or have registered via an access code will be able to view this content. To view this presentation, please login or select "Add to Cart" and proceed to checkout.