SW 1463

KAN0438757 Efficiently Reduces PFKFB3 Expression in Colorectal Cancer Cells, without Reducing Its Transcriptional Regulation

Since glycolysis is a common feature of cancer, targeting glycolysis is an attractive anti-cancer strategy. One driver of glycolysis in both cancer cells and in cells of the tumor microenvironment is PFKFB3. Oilverira et al. analyzed PFKFB3 expression in rectal and colon tumors and its prognostic significance, and investigated the effects of a novel PFKFB3 inhibitor named KAN0438757 on CRC cells and patient-derived intestinal organoids.

To test whether KAN0438757 (Fig. 1A) decreased the protein level of PFKFB3, human umbilical vein endothelial cells (HUVEC) and CRC cell lines (HCT-116, HT-29, and SW-1463) were treated with KAN0438757 (10, 25, or 50 μM) for 12 h. Immunoblot analysis revealed that KAN0438757 treatment decreased PFKFB3 protein expression in HCT-116, SW-1463, and HUVECs in a dose-dependent manner (Fig. 1B, C). HCT-116 cells were more susceptible to inhibition of PFKFB3 than SW-1463 cells, but interestingly HT-29 cells upregulated PFKFB3 at 50 μM. To determine if these effects were mediated through changes at the transcriptional level, HCT-116 and HT-29 cells were treated with KAN0438757 (10 or 25 μM) for 4-48 hours, and expression levels of PFKFB3 mRNA were measured by RT-PCR (Fig. 1D). Primary, low passage HUVECs were also treated with KAN0438757 because they are highly glycolytic. Overall, KAN0438757 did not reliably decrease PFKFB3 mRNA levels in HCT-116 cells or HUVECs. In contrast, there was a significant increase in PFKFB3 mRNA in HT-29 cells treated with KAN0438757 for 12 hours (10 and 25 μM), which did not correlate with protein levels (Fig. 1B, C). Taken together, these results suggest that KAN0438757 decreases PFKFB3 protein levels through a mechanism independent of transcriptional repression.