Altered chromosome 6 in immortal human fibroblasts.
Altered chromosome 6 in immortal human fibroblasts.
复制标题
永生人类成纤维细胞中的 6 号染色体发生了改变。
DOI：
10.1128/mcb.12.5.2273-2281.1992
复制
发表时间：
1992
影响因子：
5.3
通讯作者：
Ozer,HL
中科院分区：
文献类型：
--
作者：
Hubbard-Smith,K;Patsalis,P;Pardinas,JR;Jha,KK;Henderson,AS;Ozer,HL
人二倍体成纤维细胞在体外具有有限的寿命，并且自发永生化是极其罕见的事件。我们利用来源缺陷型猿猴病毒40基因组转化人二倍体成纤维细胞，开发了一系列遗传匹配的永生细胞系，以分析永生化。将一种永生化前的病毒（SVtsA/HF-A）与其未克隆和克隆的永生化衍生物（AR 5和HAL）进行比较，未能发现任何涉及猿猴病毒40基因组的重大改变。然而，核型分析表明，所有的永生细胞系在这一系列的染色体6的改变，涉及到6 q21远端部分的损失。DNA分析证实了核型分析。Southern分析表明，在永生细胞中仅保留了6 q上的三个原癌基因位点（ros 1，c-myb和mas 1）的一个拷贝。聚合酶链反应分析6 q22（D 6S 87）微卫星多态性显示杂合性丢失。此外，观察到c-myb（6 q22 -23）的表达升高。我们推测，该地区和/或远端6 q21发挥了永生化的作用，与生长抑制基因的存在相一致。
Human diploid fibroblasts have a limited life span in vitro, and spontaneous immortalization is an extremely rare event. We have used transformation of human diploid fibroblasts by an origin-defective simian virus 40 genome to develop series of genetically matched immortal cell lines to analyze immortalization. Comparison of a preimmortal transformant (SVtsA/HF-A) with its uncloned and cloned immortalized derivatives (AR5 and HAL) has failed to reveal any major alteration involving the simian virus 40 genome. Karyotypic analysis, however, demonstrated that all of the immortal cell lines in this series have alterations of chromosome 6 involving loss of the portion distal to 6q21. The karyotypic analysis was corroborated by DNA analyses. Southern analysis demonstrated that only one copy of three proto-oncogene loci (ros1, c-myb, andmas1) on 6q was retained in immortal cells. Polymerase chain reaction analysis of the microsatellite polymorphism at 6q22 (D6S87) showed loss of heterozygosity. In addition, elevated expression of c-myb(6q22-23) was observed. We hypothesize that the region at and/or distal to 6q21 plays a role in immortalization, consistent with the presence of a growth suppressor gene.