| Co-Investigator(Kenkyū-buntansha) |
NAKAYAMA Toshitake Keio Univ. School of Med., Associate, 医学部, 助手 (90189077)
NISHIGUCHI Iku Keio Univ. School of Med., Associate, 医学部, 助手 (20198451)
ITO Hisao Keio Uni., School of Med. Asso. Prof., 医学部, 助教授 (20095574)
尾川 浩一 慶應義塾大学, 医学部, 助手 (00158817)
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| Budget Amount *help |
¥6,900,000 (Direct Cost: ¥6,900,000)
Fiscal Year 1990: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1989: ¥5,600,000 (Direct Cost: ¥5,600,000)
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| Research Abstract |
This study was performed to establish the predictive assay method of radiotherapy for biopsy specimens from cancer patients. The first part of the study was to develop the easy and quick method to determine the cell survival after radiation. In this study, staining densitometry was applied. However, before the staining densitometry method can be accepted as the useful method, the results obtained by this method must be compared with those by the widely used colony forming method. The established cell lines (HeLa, RMUG, IMR, GOTO) grown in F10 medium with 10% fetal bovine serum were used for this study. Appropriate number of cells (10^2-3X10^3) were spread in the 24-well microplates, irradiated with different doses (0-8Gy) and cultured for about one week in the CO_2 incubator at 37C. They were stained by the 0.4% crystal violet after the culture period. Taking the transparent images of the stained well on the light source with the CCD camera, the images were collected with the matrix si
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ze 64X64, and the integrated optical density of the entire surface of each well was determined by the personal computer with our original program. As the number of cells in the well should be related with its staining density, survival was calculated by comparing the ratio of optical densities of irradiated cells with uniradiated controls. The survival curves by the densitometry method showed good correlations with those by the colony forming method. GOTO was most radiosensitive and RMUG, IMR, HeLa were less sensitive in this order. Using this method, it is able to predict the intrinsic radiosensitivity easily in a short period, even if the cells do not form colonies. As the survivals are calculated by the computer, they are reproducible without the influence of the experience of the testers.
The primary cell cultures were tried with biopsy sepcimens from head and neck cancer patients and gynecologic cancer patients. The cell survivals were calculated for these cells with the staining densitometry method. The radiosensitivity for these cells distributed in the wide range. This result suggested that the cancer cells taken from the same organ and with same histology had different radiosensitivity from the cells to the cells. However, there was no clear correlation between the outcomes of clinical radiotherapy and cell radiosensitives, because of the short observation period after radiotherapy and small number of testing.
We must try to continue more studies to fulfill the purpose. Less
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