Poot M, Hoehn H, Bogdahn U, Otto F
Department of Human Genetics, University of Würzburg, Germany.
Melanoma Res. 1992 Nov;2(4):241-6. doi: 10.1097/00008390-199211000-00004.
A model system for testing the efficacy of chemotherapy protocols for metastatic melanoma was established using cell cultures from two brain and three lymph node metastases of melanoma from five different patients. Continuously growing cultures which were positive for tyrosinase activity were analysed regarding their proliferation rate by continuous bromodeoxyuridine (BrdU) labelling and subsequent Hoechst-33258/ethidium bromide flow cytometry. Melanoma cell cultures exhibit a strong sensitivity to BrdU: at 5% oxygen, 50% growth inhibition is attained with 360 +/- 130 microM BrdU (range: 130-520; n = 11) vs 650 +/- 50 microM BrdU (n = 3) for diploid human fibroblasts and 570 +/- 20 microM BrdU (n = 6) for human lymphoid cell lines. Moreover, BrdU sensitivity of melanoma cells is clearly oxygen dependent: 50% growth inhibition at 200 +/- 55 microM (range: 65-400 microM) for 20% oxygen vs 360 +/- 130 microM BrdU for 5% oxygen. The cell cycle kinetic mechanism of BrdU-induced growth inhibition is accumulation of cells in the first cycle G2 phase. On the basis of these results we suggest testing BrdU in chemotherapy protocols for the treatment of metastatic melanoma.
利用来自5名不同患者的2个脑转移瘤和3个淋巴结转移瘤的细胞培养物，建立了一个用于测试转移性黑色素瘤化疗方案疗效的模型系统。对酪氨酸酶活性呈阳性的持续生长培养物，通过连续溴脱氧尿苷（BrdU）标记及随后的Hoechst-33258/溴化乙锭流式细胞术分析其增殖率。黑色素瘤细胞培养物对BrdU表现出很强的敏感性：在5%氧气浓度下，360±130微摩尔BrdU（范围：130 - 520；n = 11）可达到50%的生长抑制，而二倍体人成纤维细胞为650±50微摩尔BrdU（n = 3），人淋巴细胞系为570±20微摩尔BrdU（n = 6）。此外，黑色素瘤细胞对BrdU的敏感性明显依赖于氧气：在20%氧气浓度下，200±55微摩尔（范围：65 - 400微摩尔）可达到50%的生长抑制，而在5%氧气浓度下为360±130微摩尔BrdU。BrdU诱导生长抑制的细胞周期动力学机制是细胞在第一个周期的G2期积累。基于这些结果，我们建议在转移性黑色素瘤的化疗方案中测试BrdU。
