[Comparative analysis of herpes simplex virus thymidine kinase gene expression potentiation via HIV-1 Tat-TAR-system and cancer-specific promoters in p53(+) and p53(-) cells].
Tumor-specific promoters are predominantly active and ensure expression of the gene under control exclusively in cancer cells. However, a low activity of the promoters is an essential disadvantage for their therapy usage. To achieve a higher expression level of the therapeutic gene, herpes simplex virus thymidine kinase (HSV-tk), the Tat-TAR-system being utilized by HIV-1 for increasing own gene expression was developed. A potentiating activity of tat gene under control of two different cancer-specific gene (human survivin gene and human telomerase reverse transcriptase) promoters for increasing of the HSV-tk gene expression being regulated by TAR-element was evaluated, and activity of the cancer-specific promoters in the Tat-TAR-system was compared. Co-transfection of the cells with the both constructions led to the tat protein synthesis and its affect the HIV-1 TAR-element. An expression level of HSV-tk gene ensured by the both promoters in the binary system was close to that for strong non-specific cytomegalovirus (CMV) promoter. Enzymatic activity of HSV-tk protein in cells having both elements of Tat-TAR-system was two orders of magnitude higher than that in the cells transfected with HSV-tk gene under control of the cancer-specific promoter. Notably, the effect was independent of p53-status of transfected cells: HSV-tk expression level was almost the same in p53(+) and p53(-) cells. The obtained results show that system may be used for therapy of different cancer types both p53-defective and p53-positive ones inhibiting cancer-specific promoters activity.
Mingaleeva
R N
RN
Chernov
I P
IP
Kopantsev
E P
EP
Zavalova
L L
LL
Sass
A V
AV
Sverdlov
E D
ED
D045744:Cell Line, Tumor; D015870:Gene Expression; D015497:HIV-1 / Q000235:genetics*; D006564:Herpesviridae / Q000201:enzymology* / Q000235:genetics; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins / Q000235:genetics*; D009369:Neoplasms / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics*; D011401:Promoter Regions, Genetic; D016601:RNA-Binding Proteins / Q000235:genetics*; D000077022:Survivin; D019098:Telomerase / Q000235:genetics*; D013937:Thymidine Kinase / Q000096:biosynthesis* / Q000235:genetics; D016159:Tumor Suppressor Protein p53 / Q000235:genetics* / Q000378:metabolism; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000235:genetics*; D000099253:Trans-Activation Responsive RNA-Binding Protein
D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D009687:Nuclear Proteins; D016601:RNA-Binding Proteins; D000077022:Survivin; D019098:Telomerase; D013937:Thymidine Kinase; D016159:Tumor Suppressor Protein p53; D054322:tat Gene Products, Human Immunodeficiency Virus; C107489:BIRC5 protein, human; C071598:TARBP1 protein, human; D000099253:Trans-Activation Responsive RNA-Binding Protein; C495901:TP53 protein, human; C509186:TERT protein, human