DNA Histogram of Neuroblastoma Flow-microfluorometric Study

Une Yoshie, Sasaki Fumiaki, Sano Hidekazu, Hata Yoshinobu, Uchino Junichi, Kasai Yoichi

doi:10.11164/jjsps.17.2_279

Eight neuroblastomas and 1 ganglioneuroma of human subjects, 1 serial transplanted neuroblastoma in nude mice and C-1300 mouse neuroblastoma cell line were studied on its cell kinetics by flow-microfluorometry (FMF). The specimens resected from stage IV neuroblastoma patients and nude mouse were freezen in liquid nitrogen. The tumor of about 1 cm^3 block was minced and tumor cell suspensions were prepared by treatment with 0.25% Trypsin solution. Following fixation by 50% ethanol, Propidium Iodide staining was carried and DNA histogram was obtained by FMF (4800A biophisics U.S.A.). DNA histograms were divided into three different types ; unimodal, bimodal and intermediate types. Furthermore, cell population ratio of near 2C compartment were calculated. About 4×10^3 to 3×10^4 of cells were measured. Control cells were chicken red blood cells and human lymphocytes which showed 2C unimodal pattern. Five of 8 cases showed near 2C unimodal pattern. The other neuroblastomas and a ganglioneuroma were with intermediate pattern. Serial transplanted tumor in nude mice was with bimodal pattern. C-1300 mouse neuroblastoma cell line was with multimodal pattern. The specimens excised after high dose cyclophosphamide therapy showed relatively low population of 2C compartment cells compared with non-treated cases. It is still not clear how cell cycle of neuroblastoma changes by various therapeutic methods. This cytofluorometric analysis is expected to make therapeutic schedules more reasonable and effective.

DNA Histogram of Neuroblastoma Flow-microfluorometric Study

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