Published in Cancer Detection and Prevention 2000; 24(Supplement 1).

Anticancer effect of nitric oxide and therapeutic drugs on human hematological malignant cells: Analysis by in vitro monitoring using MTT assay and flow cytometry

M Tsumori MD PhD 1, J Tanaka MD PhD 1, K Koshimura MD PhD 1, M Kawaguchi PhD 1, Y Murakami MD PhD 1, Y Kato MD PhD 1

1 First Div, Dept Medicine, Shimane Medical University, Izumo, Japan,

AIMS: There have been several reports that NO has cytotoxic effects on human cell lines of leukemia and lymphoma, suggesting that NO donors can be a chemotherapeutic agent. We investigated the possibility of NO as a novel anticancer agent using primally isolated human malignant cells. METHODS: Malignant cells were obtained from venous blood, bone marrow or ascites of hospitalized patients with malignant lymphoma (n=3), multiple myeloma (n=2), acute myelogenic leukemia (n=8) and myelodysplastic syndrome (n=1). Normal leukocytes were collected from 4 healthy volunteers. After washing, collected cells were layered on Histopaque-1077 (Sigma). After centrifugation, the cells in the opaque interface was washed, plated (2-4 x 106/ml) and cultured in RPMI 1640 supplemented with 10% fetal calf serum for 3-5 days before experiments. The cells were added with anticancer drugs or sodium nitroprusside (SNP) and cultured for 3 days. Total viable cell number was measured with MTT assay and subpopulation of viable cells was analyzed by flow cytometry. RESULTS: SNP was sensitive to lymphoma cells of 2 of 3 examined patients but resistant to myeloma cells of one patient and leukemia cells of 4 patients. Prior to chemotherapy, most of anticancer drugs were sensitive to malignant cells, whereas some drugs were insensitive after chemotherapy. In contrast, SNP was sensitive to lymphoma cells after the therapy. Peripheral lymphocytes of healthy volunteers and patient with lymphoma were resistant to anticancer drugs and SNP. CONCLUSIONS: SNP was effective in lymphoma cells, but not in myeloma cells, leukemia cells or normal lymphocytes.

KEY WORDS: oxidative stress, apoptosis.

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Paper presented at the International Symposium on Impact of Biotechnology on Cancer Diagnostic & Prognostic Indicators; Geneva, Switzerland; October 28 - 31, 2000; in the section on novel therapies.