Article Figures & Data
Figures
- FIGURE 1.
Effect of OA pretreatment on specific binding of 111In-pentetreotide in cells. Binding of sst was assessed in cells pretreated with OA, 5 nmol/L, for 2 wk. Binding conditions included incubation with diagnostic dose (8.6 pmol/L) (A) or with therapeutic dose (510 pmol/L) (B) of 111In-pentetreotide in IMR-32 cells (sst2 positive) or in PANC-1 cells (sst2 negative). Treatments were terminated at 48 h, at 1 min, or not at all (no wash) before incubation with 111In-pentetreotide. Untreated control cells were also assessed for binding. Within each cell type, no statistically significant differences in specific binding of 111In-pentetreotide were found between control group and any pretreated cell group.
- FIGURE 2.
Quantitative PCR analysis of sst2 and somatostatin in IMR-32 cells treated with OA. IMR-32 neuroblastoma cells were cultured in presence of OA, 10 nmol/L or 1 pmol/L, for either 1 d or 2 wk. Untreated IMR-32 cells and TT cells were cultured as controls. Cells were then harvested, and RNA was isolated and converted to cDNA. Quantitative PCR was performed using primers and probes specific for sst2 (A) and somatostatin (B), with 18S rRNA as a control in each well. Results are shown as mean ± SD, relative to sst2 expression in IMR-32 cells and somatostatin expression in TT cells (n = 3).
Tables
Gene 5′ primer 3′ primer Probe Positive tissue* sst 5′-TGAGGCTTGA 5′GCCAGCTTTG FAM-TGCTAACTCAAAC TT (carcinoma, thyroid, medulla) GCTGCAGAGAT-3′ CGTTCTCG3′ CCGGCTATGGCAC-TAMRA sst1 5′-CCCAGAACGGG 5′AGGCACACCA FAM-AGGGCAGCGCCA SKR1 (SKNSH transfected with sst1) ACCTTGA-3′ CGGAGTAGA3′ TCCTGATCTCTT-TAMRA sst2 5′-CCAACACCTCA 5′TTGCCACACA FAM-CGTACTATGACCTGA TT (carcinoma, thyroid, medulla) AACCAGACAGA-3′ ACCCAATGAT3′ CAAGCAATGCAGTCCT-TAMRA sst3 5′-CTGCTGGGTAA 5′GAGGATGTAG FAM-TCTATGTGGTCCTG sst3/pcDNA3 CTCGCTGGT-3′ ACGTTGGTGA3′ CGGCACACG-TAMRA sst4 5′-GCAGGCGCT 5′GCATCAAGGC FAM-CAGAGCACAAAGAC sst4/pcDNA3 CGGAGAAG-3′ TGGTCACGA3′ GACCACGACCAT-TAMRA sst5 5′-CATCCTCTCC 5′GGAAGCTCTG FAM-CCCGTCCTCTACGG sst5/pcDNA3 TACGCCAACAG-3′ GCGGAAGTT3′ CTTCCTCTCTGA-TAMRA ↵* Positive control tissues were used to validate design of primers and probes.
