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首页> 外文期刊>Molecular and Cellular Biology >Transfer of Sensitivity to Tumor Promoters by Transfection of DNA from Sensitive into Insensitive Mouse JB6 Epidermal Cells
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Transfer of Sensitivity to Tumor Promoters by Transfection of DNA from Sensitive into Insensitive Mouse JB6 Epidermal Cells

机译：通过将DNA从敏感基因转染到不敏感的小鼠JB6表皮细胞中，将敏感性转移至肿瘤启动子

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Sensitivity to promotion of transformation by tumor promoters in mouse epidermal JB6 cells appears to have a genetic basis since the phenotypes of both promotable and nonpromotable JB6 cells derived from a common parent line are stable. Hybridization of promotable (P+) and nonpromotable (P?) cells previously indicated that promotability appears to behave as a dominant trait. These results suggest that it should be possible to find DNA sequences which specify sensitivity to promotion of anchorage independence by 12-o-tetradecanoyl-phorbol-13-acetate (TPA). Cellular DNA isolated from one of two P+ lines, JB6 Cl 41 or JB6 Cl 22, was CaPO4 precipitated and used to transfect the P? cell line JB6 Cl 30. At 7 days posttransfection, the cells were suspended in agar with or without TPA at 1.6 × 10?8 M and assayed 10 days later for TPA-dependent colony formation. Untreated or Cl 30 DNA-treated P? JB6 Cl 30 cells yielded 40 to 50 colonies per 105 cells. In contrast, transfection of Cl 30 cells with “P+ DNA” derived from either Cl 41 or Cl 22 yielded 200 to 500 TPA-induced colonies per 105 cells, or a five- to eightfold enhancement of promotability. The enhanced promotability obtained after transfection with P+ DNA was stable, as judged by the retention of promotability for at least eight passages in cell lines derived from TPA-induced agar colonies. Other transfectants showed irreversible transformation by TPA, as observed in the parental P+ lines. When NIH 3T3 cells instead of the putative preneoplastic JB6 Cl 30 cells were used as recipients for transfection of P+ DNA, no evidence for acquisition of promotability was obtained. P? JB6 Cl 25, like Cl 30, also permitted expression of transfected P+ DNA. These results suggest that sensitivity to phorbol ester promotion of transformation in JB6 cells is determined by DNA sequence(s) present in the P+ DNA and requires recipient cells of the appropriate phenotype for expression.
机译：小鼠表皮JB6细胞中通过肿瘤启动子促进转化的敏感性似乎具有遗传基础，因为源自共同亲本系的可促进和不可促进JB6细胞的表型都是稳定的。以前，可促销（P + ）和不可促销（P ？）细胞的杂交表明，可促销性似乎是一个主要特征。这些结果表明，应该有可能找到对12- o -十四烷酰基-佛波醇-13-乙酸酯（TPA）促进锚定独立性敏感的DNA序列。从两个P + 系之一JB6 Cl 41或JB6 Cl 22中分离出的细胞DNA沉淀了CaPO 4 并用于转染P ？细胞系JB6 Cl30。转染后7天，将细胞悬浮在有或没有TPA的琼脂中，浓度为1.6×10 M，并在10天后检测TPA依赖性菌落的形成。未经处理或经Cl 30 DNA处理的P ？ JB6 Cl 30细胞每10 5 细胞可产生40至50个克隆。相反，用源自Cl 41或Cl 22的“ P + DNA”转染Cl 30细胞每10 5 细胞产生200至500 TPA诱导的集落，或者可推广性提高了五到八倍。用T + DNA转染后获得的增强的促进能力是稳定的，这可以通过在TPA诱导的琼脂菌落衍生的细胞系中至少保留8代的促进能力来判断。如在亲本P + 品系中观察到的，其他转染子显示出TPA不可逆的转化。当NIH 3T3细胞代替假定的肿瘤前JB6 Cl 30细胞被用作P + DNA转染的受体时，没有获得促进活性的证据。 P ？ JB6 Cl 25与Cl 30一样，也允许表达转染的P + DNA。这些结果表明，对JB6细胞中佛波酯促进转化的敏感性是由P + DNA中存在的DNA序列确定的，并且需要具有适当表型的受体细胞才能表达。

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