Targeted drug delivery using an aptamer against shared tumor-specific peptide antigen of MAGE-A3

Figures & data

Figure 1. Comparison of bio-distribution of tail vein- and in situ-injected Cy5-ThioAp52 and Cy5-ThioControl probe in AsPC-1 xenograft mouse model. In vivo real-time imaging with tail vein-injected Cy5-ThioAp52 (a) or Cy5-ThioControl (b) at 0, 5, 60, 240, and 360 min post-injection, and with in situ-injected Cy5-ThioAp52 (c) or Cy5-ThioControl (d) (yellow circle at the right flank) at 0, 5 min, 4 h, 1, 2, 3, 4 and 5 d post-injection. Color scale applies to all figures. For each treatment group, results obtained with individual mice were consistent, and images from one mouse were shown

Figure 2. Binding assay of ThioAp52-Dox complex. Four pairs of cultured cancer cells (a, c, e, g) and corresponding noncancerous cells (b, d, f, h) were assayed with flow cytometry after 6 h treatment of ThioAp52-Dox (5.0 µM, green line), Dox (5.0 µM, red line), or control (culture medium only, black line). Cell lines: a) AsPC-1, b) hTERT-HPNE, c) Cal-27, d) OMF, e) MCF-7, f) MCF-10A, g) SK-MEL-28, h) Hs895.Sk

Figure 3. Uptake of FITC-ThioAp52-Dox by skin cells. Confocal laser scanning microscopy images of SK-MEL-28 (Rows I) and Hs895.Sk cells (Rows II) treated with 2.5 µM FITC-ThioAp52-Dox for 2 h (a) or 6 h (b). Columns 1) transmittance, 2) DAPI (blue), 3) FITC (for FITC-ThioAp52) (green), 4) Dox (red), and 5) superimposed image of 1) ~4). Yellow fluorescence in 5) indicates the localization of FITC-ThioAp52-Dox. Scale bar = 10 µm applies to all figures

Figure 4. Targeting of FITC-ThioAp52-Dox to cancer cells. Confocal laser scanning microscopy images of three pairs of cultured cancer cells (Columns I) and corresponding normal/noncancerous cells (Columns II) after 2 h treatment of 2.5 µM FITC-ThioAp52-Dox or Dox. Cell lines: A. Pancreatic, I) AsPC-1, II) hTERT-HPNE; B. Oral, I) Cal-27, II) OMF; C. Breast, I) MCF-7, II) MCF-10A. All micrographs are merged images of transmittance, DAPI, FITC, and Dox (See Figure S3-S6 for images of individual channels). Yellow fluorescence in Columns I indicates the localization of FITC-ThioAp52-Dox complex. Scale bar = 10 µm applies to all figures

Table 1. Uptake analysis of FITC-ThioAp52-Dox^a

Cell line	FITC-ThioAp52^b	Dox^b	Ratio^c
AsPC-1	21.3 ± 5.5	19.0 ± 5.1	1.12 ± 0.05
hTERT-HPNE	12.4 ± 2.0	14.7 ± 3.4	0.84 ± 0.09
Cal-27	19.5 ± 4.2	16.7 ± 3.5	1.15 ± 0.05
OMF	2.1 ± 0.9	7.3 ± 3.5	0.30 ± 0.09
MCF-7	12.3 ± 4.5	12.1 ± 3.7	1.01 ± 0.09
MCF-10A	9.2 ± 2.2	10.6 ± 2.1	0.84 ± 0.12
SK-MEL-28	13.4 ± 6.5	11.4 ± 5.1	1.15 ± 0.16
Hs895.Sk	1.6 ± 0.6	4.8 ± 1.6	0.36 ± 0.15

^aCells were cultured for 48 h, and the adherent cells were treated with 2.5 µM FITC-ThioAp52-Dox for 2 h and prepared for confocal microscopy quantitation of fluorescence signals.

^bMean fluorescence intensity per cell (arbitrary unit, average ± standard deviation, n = 100).

^cRatio of mean fluorescence intensity per cell, FITC-ThioAp52:Dox (average ± standard deviation, n = 100).

Figure 5. Cytotoxicity of ThioAp52-Dox. Cell viability assay in four pairs of cancer cells (

) and corresponding normal/noncancerous cells (

) after 48 h incubation with medium only (Control), or with medium containing 5.0 µM of ThioAp52 (Ap), free doxorubicin (Dox), or ThioAp52-Dox (Ap-Dox). A) AsPC-1 and hTERT-HPNE, B) MCF-7 and MCF-10A, C) Cal-27 and OMF, D) SK-MEL-28 and Hs895.Sk. Asterisks indicate the results of cancer and noncancerous cells are significantly different (p< .004)