Iartseva N M, Artsybasheva I V, Fedortseva R F, Ignatova T N
Tsitologiia. 1994;36(12):1213-24.
Seven spontaneously transformed cell lines LREC of rat embryo fibroblasts were obtained by an originally elaborated cloning technique (method 2T7). Six lines (1-6) were obtained from Rattus norvegicus cells, and one line (LREC-7) from Wistar rats. Method 2T7 was based on a serial propagation of rat embryo cells, brought to a "crisis" stage under conditions of a higher cell density and followed by the appearance of actively proliferating cell clones. These lines were analysed cytogenetically at the earliest stages using the Giemsa G-banding technique. In the karyotypes of six LREC (1-6) lines two abnormal chromosomes 7 were revealed: one marker chromosome M1-t(7; 19) results from translocation between chromosomes 7 and 19, the other marker chromosome M2--del(7) is a result of deletion of the second homolog of chromosome 7 in the q11.2 q22.1 loci; besides an extra normal homolog of chromosome 7 was revealed. There are only two marker chromosomes M2 in the LREC-7 line karyotype. Cells of LREC (1-3) lines could be transformed from the immortalized stage to the malignant one by the 30-45th passages. The cells of LREC (4, 7) lines became malignant at the 10-8th passages, resp. The rearrangements of chromosome 7 are supposed to be specific for LREC lines obtained by our method. A hypothesis is put forward that the translocation of chromosome 7 may play an important role for the immortalization of the rat embryo cells. The deletion of chromosome 7 may be associated with a malignant transformation of cells, as it is possible that the deleted loci have a recessive oncogene. Method 2T7 allows to obtain constantly spontaneously transformed cell lines of rat embryo cells with the least abnormal karyotype.
通过最初精心设计的克隆技术(2T7方法)获得了7个大鼠胚胎成纤维细胞自发转化细胞系LREC。其中6个系(1 - 6)来自褐家鼠细胞,1个系(LREC - 7)来自Wistar大鼠。2T7方法基于大鼠胚胎细胞的连续传代培养,在较高细胞密度条件下使其进入“危机”阶段,随后出现活跃增殖的细胞克隆。在最早阶段使用吉姆萨G显带技术对这些细胞系进行了细胞遗传学分析。在6个LREC(1 - 6)系的核型中发现了两条异常的7号染色体:一条标记染色体M1 - t(7; 19)是7号和19号染色体之间易位的结果,另一条标记染色体M2 - del(7)是7号染色体第二个同源染色体在q11.2 q22.1位点缺失的结果;此外还发现了一条额外的正常7号同源染色体。LREC - 7系核型中只有两条标记染色体M2。LREC(1 - 3)系的细胞在传代30 - 45次时可从永生化阶段转变为恶性阶段。LREC(4, 7)系的细胞分别在传代10 - 8次时变成恶性细胞。7号染色体的重排被认为是我们通过该方法获得的LREC细胞系所特有的。提出了一个假说,即7号染色体的易位可能对大鼠胚胎细胞的永生化起重要作用。7号染色体的缺失可能与细胞的恶性转化有关,因为缺失的位点可能含有一个隐性癌基因。2T7方法能够获得核型异常最少的大鼠胚胎细胞持续自发转化细胞系。