Flow-Cytometric Determination of Tumor Cells in Lymph Nodes<footref rid="foot01">¹</footref>

Abstract

In solid tumors, metastasis occurs through the dissemination of tumor cells in the bloodstream and the lymphatic system. In particular, lymph node infiltration gives useful prognostic information and represents one of the most important factors for selecting the type of clinical treatment in disease management. Furthermore, the analysis of lymph node infiltration has become important for identifying patients with breast cancer or malignant melanoma who may be candidates for regional lymph node dissection. Tumor cells in lymph nodes are currently identified in tissue sections using morphological and immunohistochemical analyses, but these approaches are time-consuming, and micrometastases may escape detection. The aim of the present study was to define the potential of a flow cytometric (FCM) determination based on cell size and autofluorescence to shorten the time required for lymph node analysis. The sensitivity of the FCM approach, defined on mixtures of tumor cells from established cell lines and peripheral blood lymphocytes (PBL_s) at different concentrations, was 1 tumor cell/1,000 PBL_s. FCM analysis was performed on 89 lymph nodes, 29 from breast, 41 from lung and 19 from colon cancer patients. Agreement between FCM and morphological results, used as gold standard, was observed in 83% of the cases, and there was a 90% sensitivity to the FCM approach for each tumor type. Disagreement was observed for 15 lymph nodes and was due, in the majority of cases (80%), to FCM-positive and morphologically negative results. A large number of patients and a more accurate pathological examination of consecutive histological sections of lymph nodes are needed to further evaluate the validity of the FCM approach.

Affinity of the alpha4–beta1 integrin-targeting peptide LLP2A to canine lymphoma

“…This autofluoresence may be caused by increased aerobic energy metabolism due to differing levels of NAD(P)H and flavin coenzymes between normal and neoplastic lymphocytes (Pantanelli et al, 2009). Lymphoma cells are generally larger than non-neoplastic lymphocytes, and contain cytoplasmic granules, which may also contribute to autofluorescence (Zoli et al, 2002). …”

Comparison of Mastoscopic and Conventional Axillary Lymph Node Dissection in Breast Cancer: Long-term Results From a Randomized, Multicenter Trial

“…Other groups have reported similar results, and most surgeons will not omit axillary drainage after MALND. 5,7 The drainage flow of CALND is reported to be 322 to 447 mL (range, 23-1100 mL). 40,41 In the present study, the overall flow in the MALND group was 120.91Ϯ39.22 mL over 4.04Ϯ1.29 days, slightly lower than that in the CALND group (150.29Ϯ66.92 mL over 5.29Ϯ2.01 days; Pϭ.08 for drainage and Pϭ.23 for duration).…”

“…As a result, the blood vessels and lymphatic tubes of tumor drainage, which are still patent and draining, can result in the mechanical stimulation of tumor cell metastases via blood or lymph. [6][7][8] Finally, patients are placed supine during breast cancer surgery, and the operating view of surgeons is from up to down in CALND, which restricts the axillary exposure. Although this approach is still convenient for dissecting the outer part of the axillary vein in the lateral side of the minor pectoral muscle (level I lymph node), the thoracodorsal nerve and vessels, and the long thoracic nerve, it is difficult to manipulate the major and minor pectoral muscles inward for dissecting level II lymph nodes, which are located behind the minor pectoral muscle, and level III lymph nodes and Ratter lymph node, which are between the major and minor pectoral muscles.…”

Enrichment methods to detect bone marrow micrometastases in breast carcinoma patients: clinical relevance

“…The methods currently used to evaluate the spreading of micrometastases are immunocytochemistry (IC) [4,5], RT-PCR [6,7], flow cytometry [8,9], fluorescence in situ hybridization [10,11], and immunomagnetic (IM) bead enrichment [12-15]. Most of these methods rely on the expression of epithelial markers on the membrane or the cytoskeleton of carcinoma cells collected from blood or BM after density gradient centrifugation (Ficoll) of the mononuclear cell (MNC) fraction.…”