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首页> 美国卫生研究院文献>Molecular and Cellular Biology >Activated v-myc and v-ras oncogenes do not transform normal human lymphocytes.
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Activated v-myc and v-ras oncogenes do not transform normal human lymphocytes.

机译：激活的v-myc和v-ras癌基因不能转化正常的人类淋巴细胞。

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Activated v-myc (pSV v-myc) and v-Ha-ras (GT10) oncogenes were introduced into normal human lymphocytes, NIH 3T3 fibroblasts, B-lymphoblastoid cells, and human epithelial cells, using a reconstituted Sendai virus envelope-mediated gene transfer technique. Efficient transfer of the plasmid in each cell type was demonstrable within 1.5 h of transfection by Southern blotting of extrachromosomal DNA extracts, which unexpectedly revealed that v-myc plasmid DNA was unstable in normal lymphocytes but not in the other cell types. The v-myc plasmid was stabilized when cotransfected into lymphocytes together with v-Ha-ras. The transfected v-Ha-ras plasmid was stable in all the cell types tested. v-myc plasmid expression was clearly detectable by 5 h in all cell types except human lymphocytes. Lymphocytes expressed v-myc when transfected together with v-Ha-ras. Transfected ras oncogene was efficiently expressed in all the cell types tested. Expression of the transfected genes increased at 24 and 48 h after transfection. Even though plasmid stability and expression were achieved in myc-ras-cotransfected lymphocytes, no effects on cellular DNA synthesis or immortalization were observed, in contrast to efficient transformation of NIH 3T3 fibroblasts by the same procedure. Our data suggest that efficient expression of transfected myc and ras oncogenes in normal quiescent human lymphocytes is not sufficient for the induction of cell growth and immortalization.
机译：使用重组的仙台病毒包膜介导的基因，将活化的v-myc（pSV v-myc）和v-Ha-ras（GT10）癌基因引入正常人淋巴细胞，NIH 3T3成纤维细胞，B淋巴母细胞和人上皮细胞。转移技术。通过染色体外DNA提取物的Southern印迹，可以在转染后1.5小时内证明质粒在每种细胞类型中的有效转移，这出乎意料地表明v-myc质粒DNA在正常淋巴细胞中不稳定，但在其他细胞类型中则不稳定。当与v-Ha-ras共转染到淋巴细胞中时，v-myc质粒稳定。转染的v-Ha-ras质粒在所有测试的细胞类型中均稳定。在除人类淋巴细胞以外的所有细胞类型中，均可以在5 h内检测到v-myc质粒表达。当与v-Ha-ras一起转染时，淋巴细胞表达v-myc。转染的ras癌基因在所有测试的细胞类型中均有效表达。转染后24和48小时，转染基因的表达增加。即使在myc-ras共转染的淋巴细胞中实现了质粒稳定性和表达，与通过相同程序有效转化NIH 3T3成纤维细胞相比，也未观察到对细胞DNA合成或永生化的影响。我们的数据表明转染的myc和ras致癌基因在正常的静态人淋巴细胞中的有效表达不足以诱导细胞生长和永生化。

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