Zoumpourlis V, Spandidos D A
Institute of Biological Research and Biotechnology, National Hellenic Research Foundation, Athens, Greece.
Biochem Pharmacol. 1993 Jul 6;46(1):178-81. doi: 10.1016/0006-2952(93)90363-2.
We have employed a recombinant plasmid, pBHIV1, carrying the long terminal repeat (LTR) sequences of the human immunodeficiency virus-1 (HIV-1) linked to the reporter chloramphenicol acetyl transferase (CAT) gene and to the aminoglycoside phosphotransferase (aph) gene as a selectable marker. We have introduced pBHIV1 into rat 208F and human MRCSV40TGR fibroblasts and obtained stable geneticin resistant RFBHIV1-1 and SVTGHIV1-1 cells, respectively. Both transfectant cells express CAT activity from the HIV LTR promoter. The response to the antineoplastic drug mitomycin C was studied on the LTR regulated CAT activity in both cell lines. It was found that mitomycin C at 10 micrograms/mL concentration stimulates the expression of CAT from the HIV LTR 77-fold in rat RFBHIV1-1 and 3.1-fold in human SVTGHIV1-1 cells.
我们使用了一种重组质粒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），该序列与报告氯霉素乙酰转移酶（CAT）基因以及作为选择标记的氨基糖苷磷酸转移酶（aph）基因相连。我们已将pBHIV1导入大鼠208F和人MRCSV40TGR成纤维细胞，并分别获得了稳定的对遗传霉素有抗性的RFBHIV1-1和SVTGHIV1-1细胞。两种转染细胞均从HIV LTR启动子表达CAT活性。在这两种细胞系中，研究了抗肿瘤药物丝裂霉素C对LTR调控的CAT活性的影响。结果发现，浓度为10微克/毫升的丝裂霉素C可使大鼠RFBHIV1-1细胞中HIV LTR的CAT表达刺激77倍，在人SVTGHIV1-1细胞中刺激3.1倍。
