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. 2024 Jun 17;14(10):1940–1963. doi: 10.1158/2159-8290.CD-23-1379

Figure 3.

Figure 3.

Pirin levels correlate with iron-(Fe3+) levels and with hTERT activity in colorectal cancers. A, qPCR-based mRNA analysis of Pirin gene expression in PDXs, normalized to β-actin expression in each sample. B, Image of PDX tumors developed in NSG mice. PDXs with low (0266PIR Low) and high (0186PIR High) Pirin (PIR) expression in NSG mice were treated with the iron chelator DFX or PBS for 3 weeks (twice weekly). C, Graph showing tumor volume over a 3-week study period in NSG mice injected with DFX or PBS (twice weekly). D, Telomerase activity was measured by TRAP assay in NSG mice PDXs treated with DFX or PBS for 3 weeks. Tumors were harvested after study termination, and a TRAP assay was performed. Error bars indicate the standard deviation of five biological replicates for each group. E, Graph showing levels of iron-(Fe3+) in PDX tumors (0186PIR High, 0266PIR Low) developed in NSG mice treated with PBS or DFX for 3 weeks. Error bars indicate the standard deviation of five biological replicates for each group. F, Western blot showing levels of Pirin in normal (N) and tumor (T) samples from patients with colorectal cancer used for studying telomerase and iron-(Fe3+) correlation in Fig. 1B–E. HSP90 was used as a loading control. The samples marked red did not show any correlation between Pirin and telomerase. G, Schematic illustrating the construction of tissue microarray using tumor and adjacent normal tissues from patients with colorectal cancer and staining for iron (PB staining) and Pirin (IHC) staining) in 245 patients with colorectal cancer matched normal and tumor samples. H and I, Representative images of two patient samples (tumor and adjacent normal) stained for iron with PB (H) and for Pirin with IHC (I). Iron-stained regions in tumor sections are marked by black circles. J, Pie chart showing percentages of tumor (left) and adjacent normal (right) samples showing high and low iron-(Fe3+) and low Pirin levels by PB staining and IHC, respectively. *, P < 0.05; **, P < 0.005; ***, P < 0.0001; n.s., nonsignificant.