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In Vitro Investigations of Tumor Targeting with ^99mTc-Labeled Antisense DNA

Division of Nuclear Medicine, Department of Radiology, University of Massachusetts Medical School, Worcester, Massachusetts

View larger version (14K): [in a new window]   FIGURE 1. Size-exclusion HPLC radiochromatograms of 99mTc-labeled antisense MAG3-amide/linker-DNA before (A) and after (B) addition of streptavidin-biotin-sense DNA. Almost quantitative shift to higher molecular weight after addition of complementary DNA and streptavidin shows conclusively that 99mTc is labeling DNA.

View larger version (68K): [in a new window]   FIGURE 2. Gel electrophoretogram shows increased expression of RI mRNA in ACHN cells relative to that of ß2m housekeeping gene. M = DNA standard marker.

View larger version (18K): [in a new window]   FIGURE 3. (A) Cell accumulation over time of 35S after incubation of ACHN cells with 3' end-labeled native, amine/linker-, and MAG3-amide/linker-antisense DNA compared with amine/linker-sense (control) DNA. Significantly higher accumulation at most time points is evident for antisense DNAs relative to sense DNA. (B) Efflux of 99mTc from ACHN cells after 16 h of incubation with 99mTc-labeled MAG3-amide/linker-antisense and sense DNAs. AS = antisense.

View larger version (18K): [in a new window]   FIGURE 4. (A) Cell accumulation over time of MAG3/linker-antisense and MAG3/linker-sense DNA radiolabeled with 35S or 99mTc. Significantly higher accumulation at most time points is evident for both antisense DNAs relative to both sense DNAs. Accumulation of 99mTc is lower than that of 35S when attached to antisense DNA. (B) Efflux of 35S and 99mTc from ACHN cells after 16 h of incubation with 35S- and 99mTc-labeled MAG3-amide/linker-antisense DNA. AS = antisense.

View larger version (147K): [in a new window]   FIGURE 5. Microautoradiography images at x400 magnification of ACHN cells incubated with 35S-labeled antisense and sense DNAs for 4 h (antisense [A]; sense [B]) and 24 h (antisense [C]; sense [D]).

View larger version (18K): [in a new window]   FIGURE 6. (A) Cell accumulation over time of 99mTc-labeled antisense and sense DNA in LS174T cells. These cells also express target RI mRNA. (B) Cell accumulation over time of 99mTc-labeled antisense and sense DNA in HC-2 cells (compared with RI mRNA–positive ACHN cells). These cells do not express target RI mRNA. AS = antisense.