HAMSTER-CELL LINE SUITABLE FOR TRANSFECTION ASSAY OF TRANSFORMING GENES
HAMSTER-CELL LINE SUITABLE FOR TRANSFECTION ASSAY OF TRANSFORMING GENES
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DOI：
10.1073/pnas.87.7.2409
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发表时间：
1990-04-01
影响因子：
11.1
通讯作者：
KAKUNAGA, T
中科院分区：
文献类型：
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作者：
HIGASHI, T;SASAI, H;KAKUNAGA, T
我们建立了一个亚克隆，SHOK，从GHE-L细胞系，从叙利亚仓鼠胚胎细胞原代培养的不死系，作为受体细胞系，用于检测癌基因转染。SHOK细胞几乎和NIH 3T3细胞一样容易受到许多致癌基因的影响，包括v-raf、v-Ha-ras、v-Ki-ras或活化的c-Ha-ras。SHOK对v-mos的敏感性高于NIH 3T3，但对v-fps、v-fgr、v-src、v-sis和v-abl的敏感性低于NIH 3T3。从27个人类和小鼠肿瘤中提取的dna进行了病灶形成检测，初代转染时，NIH 3T3细胞中有5个dna阳性，而SHOK细胞中有9个阳性。使用SHOK细胞作为肿瘤细胞DNA的受体，我们分离了另一个癌基因和一个在NIH 3T3细胞中难以检测到的密码子146突变的c-Ki-ras2基因。SHOK细胞自发转化率低，产生容易区分的病灶，并在转化细胞中保持稳定的核型。除了可用于筛选人类肿瘤dna外，SHOK细胞还可用于从小鼠肿瘤中分离癌基因，因为它们起源于仓鼠。
We established a subclone, SHOK, from the GHE-L cell line, an immortal line derived from a primary culture of Syrian hamster embryo cells, as a recipient cell line useful for the detection of oncogenes by transfection. SHOK cells were almost as susceptible as NIH 3T3 cells to focus formation by many oncogenes, including v-raf, v-Ha-ras, v-Ki-ras, or activated c-Ha-ras. The susceptibility of SHOK to focus formation was higher than that of NIH 3T3 for v-mos but was lower for v-fps, v-fgr, v-src, v-sis, and v-abl. When DNAs extracted from 27 human and murine tumors were tested for focus formation, 5 DNAs were positive in NIH 3T3 cells, whereas 9 were positive in SHOK cells at the primary transfection. Using SHOK cels as recipients of tumor cellular DNA, we isolated another oncogene and a c-Ki-ras2 gene mutated at codon 146 that were difficult to detect in NIH 3T3 cells. SHOK cells have a low rate of spontaneous transformation, produce easily distinguishable foci, and maintain a stable karyotype in transformed cells. In addition to being useful for the screening of human tumor DNAs, SHOK cells will be useful for the isolation of oncogenes from murine tumors because of their hamster origin.