Article Figures & Data
Figures
- Figure 1.
T, EphA2, and ceruloplasmin promoters are active in ovarian cancer. A, schematic of the promoter-driven luciferase report plasmids. B, a panel of ovarian cancer cell lines, normal ovarian epithelial cells (NOE99, NOE115, and NOE119), and fibroblasts (WI-38) were transiently cotransfected with plasmid DNA indicated and pRL-TK, the internal control. At 48 h following transfectioin, dual luciferase ratio was measured as the ratio of luciferase normalized to the Renilla luciferase internal control. The data are means of four independent experiments. Bar, SD.
- Figure 2.
T-VISA is robust in ovarian cancer cell lines. A, schematic diagram of engineered T-VISA constructs in the pGL3 backbone. B, using a panel of ovarian cancer cell lines, normal ovarian epithelial cells (NOE99, NOE115, and NOE119), and normal lung fibroblasts (WI-38), plasmid DNA and pRL-TK were transiently cotransfected with the indicated. Forty-eight hours later, dual luciferase ratio was measured and shown as relative light units (ratio) normalized to the Renilla luciferase control. The data are means of four independent experiments. Bar, SD. C, the percentage of T promoter activity normalized to that of the CMV promoter.
- Figure 3.
T-VISA transcriptionally targets transgene expression to SKOV3.ip1 tumors in vivo. A, female nude mice bearing orthotopic SKOV3.ip1 tumors were given DNA:liposome complexes containing 50 μg of plasmid DNA through tail-vein injection. Two days later, mice were anesthetized and subjected to in vivo imaging 10 min after i.p. injection of D-luciferin for 2 min. B, SKOV3.ip1 tumors of mice, as well as each of their heart/lungs from A, were subjected to ex vivo imaging. The photon signals were quantified by Xenogen's Living Imaging software (right). C, tissue specimens from tumor, as well as other organs harvested from mice in A, were measured for luciferase activity expressed as relative light units per milligram of total protein. Bars, SD; n = 3 per group. The plasmids indicated are as follows: CMV-Luc, pGL3-CMV-Luc; T-VISA-Luc, pGL3-T-VISA-Luc; Ctrl, pGL3-T-VISA. Asterisk, P < 0.05.
- Figure 4.
Cell killing activities of CMV-E1A and T-VISA-E1A in ovarian cancer cell lines and normal cells. A panel of ovarian cancer cell lines, normal ovarian epithelial cells, and normal fibroblasts were cotransfected with pUK21-T-VISA-E1A, pUK21-CMV-EIA, or a negative control pUK21-TV plus 100 ng of pGL3-CMV-Luc. The signals were imaged with the In vivo Imaging System 2 d posttransfection. The percentages of cell killing are indicated by the loss of luciferase reporter signals with the negative control set at 100%. The data are means of three independent experiments. Bars, SD.
- Figure 5.
T-VISA-E1A inhibited tumor growth in orthotopic xenograft animal model. A, nude mice bearing SKOV3.ip1-Luc tumors were i.v. injected with DNA:liposome complexes containing 15 μg of plasmid TV-E1A (▪) or control (○) at time points (arrows). The photon signals were quantified by Xenogen's Living Imaging software. n = 12 mice per group. B, survival rates of mice observed for 70 d after first treatment. C, analysis of in vivo apoptosis tissue samples of tumor, lung, and liver harvested from mice treated with either TV-E1A or control DNA-liposome complexes in A. D, the percentages of apoptotic cells from two fields of the indicated tumor specimens in B. Asterisk, P < 0.02.
- Figure 6.
Acute toxicity study on T-VISA-E1A versus CMV-E1A. Female CD-1 mice were given single doses of DNA:liposome complexes containing 50 μg plasmid DNA through the tail vein. Kaplan-Meier survival analysis (A) and serum levels of alanine aminotransferase 2 d after injection (B) were determined. Asterisk, P < 0.02.
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Volume 8, Issue 8
