Bifunctional chimeric SuperCD suicide gene -YCD: YUPRT fusion is highly effective in a rat hepatoma model

Figure 3 Molecular and functional analysis of subcutaneously grown control tumors explanted on d 18 (saline-treated MH SuperCD/MH naïve tumors). A: Agarose gel electrophoresis of SuperCD DNA-specific PCR products. Lane M: marker DNA ladder; lane MH: MH parental cell line (no transgene; negative control; PCR performed with 1 μg of DNA); lanes 1-3: PCR amplifications performed on MH SuperCD tumor tissues of animals 1, 2, 3 (PCR performed with 1–3 µg of DNA); lane V: vector control lane (PCR performed with 1 pg of pLXSN-SuperCD). Arrow: indicates 1 143-bp SuperCD-specific amplification product. B: SRB cytotoxicity assay with recultured tumor cells explanted after 18 d of subcutaneous growth. As in Figure 2, cytotoxicity was measured after 5-FC treatment for 4 d in vitro. MH naïve: negative control with MH parental cell line (no transgene; no animal passage); MH naïve, #1: MH naïve tumor explanted from a saline-treated animal (control); MH SuperCD, cell line: MH SuperCD parental cell line (control; no animal passage); MH SuperCD 1, 2, 3, tumor passage: MH SuperCD tumors explanted from saline-treated animals 1, 2, 3. All values were referred to that of the untransduced control cells and are given as percentage of surviving cells (control: 100%). All experiments were carried out in quadruplicate. Viability values plotted represent fitted medians and 95%CI.