Spandidos D A, Zoumpourlis V, Kotsinas A, Maurer H R, Patsilinacos P
Institute of Biological Research and Biotechnology, National Hellenic Research Foundation, Athens, Greece.
Genet Anal Tech Appl. 1990 Sep;7(5):138-41. doi: 10.1016/0735-0651(90)90020-g.
We constructed a recombinant plasmid, pBHIV1 carrying the long terminal repeat (LTR) of the human immunodeficiency virus 1 (HIV-1), linked to the chloramphenicol acetyl transferase (CAT) gene plasmid. Plasmid pBHIV1 also contains the aminoglycoside phosphotransferase gene as a selectable marker. We introduced pBHIV1 in rat 208F fibroblasts and obtained stable geneticin resistant RFBHIV1-1 transfectant cells. A further control used was plasmid p202A, which carries the mutant T24 H-ras1 promoter linked to the promotorless cat gene. Plasmid p202A also carries the aph gene as a selectable marker and was transfected into 208F cells to obtain stable transfectant RF202A-1 cells. Both RFBHIV1-1 and RF202A-1 cells expressed CAT activity from the HIV LTR and T24 H-ras1 promoters. The response to cis-platin, a platin derivative and hexadecyl-phosphocholine was studied on the HIV LTR and H-ras1 regulated CAT activity in RFBHIV1-1 and RF202A-1 cells. It was found that at 5 x 10(-5) M concentrations cis-platin stimulates by 22-fold the expression of CAT from the HIV LTR, whereas only a 4-fold stimulation was observed on the T24 H-ras1 promoter. Our results suggest caution against therapy including this compound at cytotoxic concentrations in the treatment of AIDS patients.
我们构建了一个重组质粒pBHIV1，它携带人类免疫缺陷病毒1型（HIV-1）的长末端重复序列（LTR），并与氯霉素乙酰转移酶（CAT）基因质粒相连。质粒pBHIV1还含有氨基糖苷磷酸转移酶基因作为选择标记。我们将pBHIV1导入大鼠208F成纤维细胞，获得了对遗传霉素具有抗性的稳定转染细胞RFBHIV1-1。另一个用作对照的是质粒p202A，它携带与无启动子的cat基因相连的突变型T24 H-ras1启动子。质粒p202A也携带aph基因作为选择标记，并被转染到208F细胞中以获得稳定的转染细胞RF202A-1。RFBHIV1-1和RF202A-1细胞均从HIV LTR和T24 H-ras1启动子表达CAT活性。研究了顺铂（一种铂衍生物）和十六烷基磷胆碱对RFBHIV1-1和RF202A-1细胞中HIV LTR和H-ras1调控的CAT活性的影响。结果发现，在5×10⁻⁵M浓度下，顺铂可使HIV LTR的CAT表达增加22倍，而在T24 H-ras1启动子上仅观察到4倍的刺激作用。我们的结果表明，在治疗艾滋病患者时，对于使用该化合物的细胞毒性浓度进行治疗需谨慎。
