IMMORTALIZATION OF NORMAL HUMAN FIBROBLASTS BY TREATMENT WITH 4-NITROQUINOLINE 1-OXIDE
IMMORTALIZATION OF NORMAL HUMAN FIBROBLASTS BY TREATMENT WITH 4-NITROQUINOLINE 1-OXIDE
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DOI：
10.1002/ijc.2910530317
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发表时间：
1993-02-01
影响因子：
6.4
通讯作者：
NAMBA, M
中科院分区：
文献类型：
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作者：
BAI, L;MIHARA, K;NAMBA, M
将来源于6周龄人胚胎的正常人成纤维细胞（OUMS-24株）转化（转化成OUMS-24 F系），并通过用4-硝基喹啉I-氧化物（4 NQO）重复处理（59次）使其永生化。在原代培养期间开始处理，并在第51群体倍增水平（PDL）时结束。在第57个PDL时（末次给药后146天），形态发生改变，上皮型细胞出现，开始生长并变得永生（现在已超过第100个PDL）。然而，未用4 NQO处理的对照成纤维细胞在第62个PDL衰老。正常人细胞的永生化需要用4 NQO进行广泛、重复的处理，这一发现表明，一般来说，人细胞的永生化涉及多种突变事件。换句话说，永生本身似乎是一个多步骤的过程。核型分析表明，许多细胞亚二倍体永生化之前，但后来染色体广泛分布在二倍体至四倍体区域。永生化细胞扩增，c-myc表达增强。双向电泳分析表明，细胞永生化后，消失的细胞蛋白质数量大于新出现的蛋白质数量。由于永生化细胞既不显示锚定非依赖性生长，也不显示致瘤性，因此它们可用于研究可能导致人类细胞中多步致癌的因素。此外，基因匹配的正常（OUMS-24）和永生化（OUMS-24 F）细胞将有助于分析与细胞死亡率和永生化相关的基因。
Normal human fibroblasts (the OUMS-24 strain), derived from a 6-week-old human embryo, were transformed (into the OUMS-24F line) and immortalized by repeated treatments (59 times) with 4-nitroquinoline I-oxide (4NQO). Treatment began during primary culture and ended at the 5 1 st population doubling level (PDL). At the 57th PDL (146 days after the last treatment), morphologically altered, epithelial-type cells appeared, began to grow and became immortal (now past the 100th PDL). However, the control fibroblasts, which were not treated with 4NQO, senesced at the 62nd PDL. The finding that extensive, repeated treatments with 4NQO are required for the immortalization of normal human cells, indicates that multiple mutational events are involved in the immortalization of human cells in general. In other words, immortalization itself seems to be a multi-step process. Karyotypic analysis showed that many cells were hypodiploid before immortalization, but that afterwards chromosomes were distributed broadly in the diploid to tetraploid regions. The immortalized cells showed amplification and enhanced expression of c-myc. Two-dimensional electrophoretic analysis showed that the number of disappearing cellular proteins was greater than the number of the newly appearing ones after the cells became immortalized. Since the immortalized cells showed neither anchorage-independent growth nor tumorigenicity, they are useful for studying factors that can contribute to multi-step carcinogenesis in human cells. In addition, genetically matched normal (OUMS-24) and immortalized (OUMS-24F) cells will be useful for analyzing the genes related to cellular mortality and immortalization.