Article Figures & Data
Figures
- Fig. 1.
Modulation of adenovirus-mediated transgene expression in ovarian cancer cells. Cells were incubated with or without agents at indicated concentrations for 24 h and infected with a luciferase-expressing virus. Results indicate luciferase expression as a percentage of expression with virus only. A–D, chemotherapeutics; E and F, histone deacetylators; G, RAS inhibitor; H and I, other agents. Error bars indicate 1 SD. Data were normalized to the number of cells. ∗, P < 0.05 versus no agent; ∗∗, P < 0.01; ∗∗∗, P < 0.001.
- Fig. 2.
Effect of agents on CAR expression and transcription. A, FACS detection of CAR expression. Results are presented as the mean fluorescence intensity obtained with the agent versus no agent (%). The dashed line indicates 100%. +, positive concordance with transgene expression data; −, negative concordance. B, effect of agents on CAR mRNA transcription. Quantitative RT-PCR was used to determine the copy number of CAR mRNA. Results were normalized for the number of cells by determination of glyceraldehyde-3-phosphate dehydrogenase mRNA copy number. Differences are not statistically significant. C, effect of combinations of agents on transgene expression. Results are displayed as percentage increase in comparison to no agent. Error bars indicate 1 SD. All data were normalized to the number of cells. ∗, P < 0.05 versus no agent; ∗∗, P < 0.01; ∗∗∗, P < 0.001.
- Fig. 3.
FACS analysis of CAR expression in ovarian cancer cell lines treated with FR901228, trichostatin A, topotecan, or etoposide. The thick black line indicates cells treated with the agent; the dotted line indicates no agent; and the thin gray line indicates the antibody control. For each panel, the bottom number (+) indicates the mean fluorescence intensity for the treated cell population, whereas the top number (−) is the untreated population.
- Fig. 4.
Detection of virus internalization after treatment with FR901228, trichostatin A, topotecan, or etoposide. Cells were incubated in the presence of the agents for 24 h. Virus was then added and allowed to internalize, and this was followed by rigorous washing and extraction of DNA from the cell fraction only. Virus copy number was detected with quantitative PCR. ∗, P < 0.05 versus no agent; ∗∗, P < 0.01; ∗∗∗ P < 0.001.
- Fig. 5.
Modulation of adenoviral transgene expression in ovarian cancer samples purified from patient ascites. Cells were suspended as spheroids and incubated for 24 h in the presence of the indicated agents followed by infection with an adenovirus coding for luciferase. Twenty-four h later, luciferase activity of the cell suspensions was determined, and the results were normalized for the amount of cells via measurement of total protein. Error bars indicate 1 SD. ∗, P < 0.05 versus no agent; ∗∗, P < 0.01; ∗∗∗, P < 0.001.
- Fig. 6.
Modulation of adenoviral transgene expression in an orthotopic murine model of peritoneally disseminated ovarian cancer. i.p. carcinomatosis was established, and 14 days later, agents were injected i.p. twice, followed by injection of adenovirus coding for luciferase. Twenty-four h later, tumors were collected (N = 4 mice/group; N = 2 tumor nodules collected/mouse). Half of each tumor was analyzed histologically to confirm a high viable tumor content, and the other half was subjected to transgene expression analysis. Data were normalized to the number of cells. ∗, P = 0.037 versus no agent.
Volume 63, Issue 4
