Objective:To analyze the drug sensitivity of human lung adenocarcinoma stem cells (LASC) to cisplatin (DDP) and carboplatin (CBP). Methods:Human lung adenocarcinomaic cells SPC-A1,AG,and CPA-Y2 were treated with DDP and CBP. The cell viability of cells was detected by CCK-8 assay. The phenotypic characteristics of drug surviving cells(DSCs)were determined by immunofluorescence staining. The LASC population was then separated by magnetic-activated cell sorting method. The LASC in DSCs was traced by using green fluorescence protein (GFP). The drug sensitivity of DSCs to DDP and CBP was analyzed.Results:The LASC exhibited the phenotypes of bronchioalveolar stem cells (BASC, OCT4~+CCSP~+SP-C~+). After mixture of CD221~+LASC with CD221~-lung adenocarcinoma differentiated cells, the DSC population showed OCT4~+BASC phenotypes. These DSCs were significantly resistant to DDP and CBP.Conclusion:LASC has a high resistance to DDP and CBP. This may be the reason for tumor recurrence after chemotherapy.

OBJECTIVETo evaluate the effect of vascular endothelia1 growth factor (VEGF) on the hematogenous metastasis of non-small cell lung cancer (NSCLC).

METHODSThe identification of lung cancer cells in the peripheral blood were carried out by cytological, immunohistocytologica1 and immunofluorecent stains respectively, following isolation of cytokeratin-expressing cells with magnetic activated cell sorting. The quantification of cancer cells in the blood was performed according to the established flow cytometric assay. The plasma VEGF was measured by commercially available ELISA kit.

RESULTSThe lung cancer cells in the blood, showing a remarkable nuclear polymorphism, expressed the epithelial marker cytokeratin and telomerase reverse transcriptase (hTERT). These cells were stained positive by an NSCLC-specific monoclonal antibody S5Al0-2, but negative by antibodies against CD34 and CD45 antigens. Using the flow cytometric assay, 44 cases (28.6%) of l54 NSCLC patients were found to have cancer cells in their blood, with the incidence of positive cases correlated with the stage of disease. The plasma VEGF level was significantly increased in NSCLC patients in comparison with healthy individuals and patients with benign pulmonary diseases. This level was correlated with the stages of disease in patients with adenocarcinoma. In patiens with cancer cells in their blood, a higher level of plasma VEGF was related with an increased number of cancer cells.

CONCLUSIONThe plasma VEGF level is increased in NSCLC patients with approximate1y one fourth to have cancer cells in the peripheral blood. In these patients, increased VEGF level promotes hematogenous tumor metastasis, as indicated by a much higher number of cancer cells in the blood.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Non-Small-Cell Lung ; blood ; pathology ; DNA-Binding Proteins ; Endothelial Growth Factors ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Immunohistochemistry ; Keratins ; analysis ; Lung Neoplasms ; blood ; pathology ; Lymphokines ; blood ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplastic Cells, Circulating ; chemistry ; pathology ; Telomerase ; analysis ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

BACKGROUNDTo investigate the clinical possibility of detection of lung cancer cells in the peripheral blood with cytokeratin and 2F7/S5A monoclonal antibody by flow cytometry (FCM) in patients with lung cancer.

METHODSFive mililiter peripheral blood was collected. The mononuclear cells were isolated by Ficoll-Hypaquegradient centrifugation. Then the cells were labeled with lung specific antibody (2F7/S5A) and antibodies against CD45, cytokeratin (CK). The quantitation of CD45 -CK +2F7/S5A + cells were performed by FCM.

RESULTSA total of 151 people were studied, including 20 healthy controls, 116 patients with lung cancer and 15 patients with benign pulmonary diseases. The phenotype of monocytes of peripheral blood was CD45 +CK -2F7/S5A -, while the phenotype of lung cancer cells was CD45 -CK +2F7/S5A +. No cancer cell was found in peripheral blood of all the healthy controls. One out of 15 cases of benign pulmonary diseases was found to have CD45 +CK -2F7/S5A - cells in peripheral blood. And 39 out of 116 patients with lung cancer were found to have circulating lung cancer cells. The sensitivity, specificity, positive predictive value and negative predictive value of detection was 33.62% , 93.33%, 97.50% and 15.38%, respectively. The average quantitation of circulating cancer cell detected was 0.12×10⁶/L.

CONCLUSIONSThere is high specificity and positive predictive value of the examination of circulating cancer cells in patients with lung cancer by FCM, which could detect trace cancer cell in peripheral blood of lung cancer. It may be valuable in diagnosis and treatment of lung cancer.