Cloning and sequencing of CATR1.3, a human gene associated with tumorigenic conversion.
Cloning and sequencing of CATR1.3, a human gene associated with tumorigenic conversion.
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CATR1.3（一种与致瘤转化相关的人类基因）的克隆和测序。
DOI：
10.1073/pnas.92.14.6409
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发表时间：
1995
影响因子：
11.1
通讯作者：
Milo,GE
中科院分区：
文献类型：
--
作者：
Li,D;Noyes,I;Shuler,C;Milo,GE
人鳞状细胞癌细胞系SCC 83 -01-82（SCC）含有H-ras和p53基因的突变，但在裸鼠中表现出非致瘤性表型。通过用诱变剂甲磺酸甲酯（MMS）处理，该细胞系可转化为具有致瘤表型的细胞系SCC 83 -01- 82 CA（CA）。这表明致瘤性可能需要导致协同肿瘤易感基因表达的其他遗传事件。为了鉴定协同基因，从致瘤性Ca细胞制备表达cDNA文库。将文库DNA转染到非致瘤性SCC细胞中，然后将转染的SCC细胞注射到裸鼠中用于选择致瘤性表型。注射后，18只小鼠中有3只发生肿瘤。从这些转染的细胞诱导的肿瘤中建立了几种新的细胞系，并命名为CATR细胞。这些细胞的肿瘤组织学和核型分析表明，它们是人类上皮细胞来源。所有CATR细胞都具有整合在其基因组中的文库载体序列。细胞系CATR 1表达了一个单一的信息，从代表一个1.3 kb的cDNA插入，是不存在的未转染的SCC细胞或MMS转化的CA细胞的集成库。通过PCR扩增逆转录的CATR 1总RNA克隆了该1.3 kb cDNA插入片段，并将其命名为CATR 1.3。CATR1.3的核苷酸序列编码79个氨基酸的肽，具有长的3'非翻译区，并且代表与由于转染的表达文库引起的致瘤转化相关的未知基因产物。
The human squamous cell carcinoma cell line SCC83-01-82 (SCC) contains mutations in both the H-ras and p53 genes, but it exhibits a nontumorigenic phenotype in nude mice. This cell line can be converted into a cell line with a tumorigenic phenotype, SCC83-01-82CA (CA), by treatment with the mutagen methyl methanesulfonate (MMS). This indicates that additional genetic events leading to expression of a cooperating tumor susceptibility gene(s) may be required for tumorigenicity. To identify the cooperating gene(s), an expression cDNA library was made from tumorigenic Ca cells. The library DNA was transfected into nontumorigenic SCC cells and the transfected SCC cells were then injected into nude mice for the selection of a tumorigenic phenotype. Tumors developed in 3 of the 18 mice after injection. Several new cell lines were established from these transfected cell-induced tumors and designated as CATR cells. Tumor histology and karyotype analysis of these cells indicated that they were of human epithelial cell origin. All the CATR cells have the library vector sequence integrated in their genome. Cell line CATR1 expressed a single message from the integrated library representing a 1.3-kb cDNA insert that was absent from untransfected SCC cells or MMS-converted CA cells. This 1.3-kb cDNA insert was cloned by PCR amplification of reverse-transcribed CATR1 total RNA and was designated CATR1.3. The nucleotide sequence of CATR1.3 encodes a peptide of 79 amino acids, has a long 3' untranslated region, and represents an unknown gene product that was associated with the tumorigenic conversion due to the transfected expression library.