Figure 1.
Western blots of VEGF isoforms proteins expression in lung cancer cells and real-time quantitative RT-PCR of VEGF isoform mRNA expression in tumor implants.
A. Western blots of VEGF isoform protein expression in cell lysate of human CL1-0 lung cancer cells transfected with single different VEGF isoform constructs. Each VEGF isoform protein comprised one glycosylated (upper) and one unglycosylated (lower) protein. Tubulin was used as an internal control. B. Quantification of VEGF mRNA expression in vivo in the tumor implants by real-time quantitative reverse transcription-PCR. The expression level of VEGF isoform was similar among CL1-0 lung cancer cell lines overexpressing one of three VEGF isoforms (p = 0.953, one-way ANOVA).
Figure 2.
Determining the core and rim regions of tumors on T2WI for analyzing quantitative maps (Ktrans, rVSI, and rVDI).
Figure 3.
Growth curves of different VEGF isofrom overexpressing tumors and the mock tumors.
Tumor growth of CL1-0 cancer cells overexpressing one of three VEGF isoforms at different time points in SCID mice, estimated using T2WI. #, VEGF189 versus VEGF121 and the mock tumors; +, VEGF165 versus VEGF121 and the mock tumors; *, comparison between different isoform-expressing and mock tumors. Differences between VEGF-overexpressing tumors and mock tumors were significant at the p<0.05 (one symbol), p<0.01 (two symbols), p<0.001 (three symbols), and p<0.0001 (four symbols) levels.
Figure 4.
The temporal Ktrans maps and values of different VEGF isofrom overexpressing tumors and the mock tumors.
In vivo temporal Ktrans map and quantitative curve for tumor xenografts of CL1-0 cancer cells overexpressing one of three VEGF isoforms at different time points, evaluated by DCE-MRI. (A) Representative Ktrans color map in the different VEGF-overexpressing and mock-transfected tumors (within dotted circle) at different times, from day 7 to day 35 postimplantation. The color ranged from blue (0/min, lowest Ktrans) to red (0.8/min, highest Ktrans). B) Quantitative analysis of Ktrans values over time in the regions of the whole tumor (upper), tumor rim (middle), and tumor core (lower). #, VEGF189 versus other isoforms and the mock tumors; +, VEGF165 versus VEGF121 and mock tumors. Differences between VEGF-overexpressing tumors and mock tumors were significant at the p<0.05 (one symbol), p<0.01 (two symbols), p<0.001 (three symbols), and p<0.0001 (four symbols) levels. (C) Ktrans values on day 35 in the tumor rim (upper) and tumor core (middle), and the ratio of Ktrans values between the tumor core and tumor rim (lower). Differences between VEGF-overexpressing tumors and mock tumors were significant at the *p<0.05, **p<0.01, ***p<0.001, and ****p<0.0001 levels.
Figure 5.
The rVDI and rVSI maps and values of different VEGF isofrom overexpressing tumors and the mock tumors.
In vivo rVDI and rVSI maps and quantitative curves for tumor xenografts of CL1-0 cancer cells overexpressing one of three VEGF isoforms, evaluated by SSCE-MRI. Representative high-resolution maps of the (A) rVDI and (B) rVSI in the different VEGF-overexpressing and mock tumors on day 36 after tumor implantation. In rVDI map, the color ranged from blue (0 S-1/3, lowest rVDI) to red (0.4 S-1/3, highest rVDI). In rVSI map, the color ranged from blue (0, lowest rVSI) to red (30, highest rVSI).Quantitative analysis of (C) rVDI and (D) rVSI in the whole tumor (upper), tumor rim (middle), or tumor core (lower). Differences between VEGF-overexpressing tumors and mock tumors were significant at the *p<0.05, **p<0.01, ***p<0.001, and ****p<0.0001 levels.
Figure 6.
The immunohistochemical staining of microvessels of different VEGF isofrom overexpressing tumors and the mock tumors.
Angiogenesis phenotypes of microvessels evaluated by immunohistochemical staining of tumor xenografts overexpressing different VEGF isoforms. (A) Immunohistochemical staining of tumor microvessels (brown color, ×100 in main panel, ×400 in the inset) in tumor xenografts. (B) The microvessel density was highest in the VEGF189-overexpressing tumors, intermediate in the VEGF165-overexpressing tumors, and lowest in the VEGF121-overexpressing tumors. (C) The number of vessels with a diameter larger than 15 µm per tumor section in the tumor xenograft was highest in the VEGF121-overexpressing tumors, intermediate in the VEGF165-overexpressing tumors, and lowest in the VEGF189-overexpressing tumors. Differences between VEGF-overexpressing and mock tumors were significant at the *p<0.05, **p<0.01, ***p<0.001, and ****p<0.0001 levels.