GANT61 exerts anticancer cell and anticancer stem cell capacity in colorectal cancer by blocking the Wnt/β‑catenin and Notch signalling pathways

Abstract

The present study aimed to assess the anticancer cell and anticancer stem cell (CSC) effects of GANT61, and its regulatory influence on the Wnt/β-catenin and Notch signalling pathways in colorectal cancer (CRC). HT-29 and HCT-116 cells were treated with 0, 2.5, 5, 10, 20 or 40 µM GANT61, after which relative cell viability and the expression of Gli1, β-catenin and Notch1, as well as the percentage of CD133 + cells, were detected. Subsequently, HT-29/HCT-116 cells and CSCs were treated with 20 µM GANT61, 10 mM of the Wnt/β-catenin pathway agonist HLY78, and 30 mM of the Notch pathway agonist JAG1 (alone or in combination), which was followed by the assessment of cell viability and apoptosis. In both cell lines, GANT61 reduced relative cell viability in a time-and dose-dependent manner, inhibited Gli1, β-catenin and Notch1 expression, and decreased the percentage of CD133 + cells in a dose-dependent manner. Furthermore, HLY78 and JAG1 were both found to improve the relative viability, while downregulating the apoptosis of untreated and GANT61-treated HT-29 and HCT-116 cells. Moreover, Wnt/β-catenin and Notch signalling pathway activity were upregulated in CSCs isolated from HT-29 and HCT-116 cells, compared with the associated control groups. GANT61 also reduced the viability of HT-29 and HCT-116 cells and increased apoptosis, whereas HLY78 and JAG1 treatment resulted in the opposite effect. Moreover, both HLY78 and JAG1 attenuated the effects of GANT61 on cellular viability and apoptosis. In conclusion, GANT61 was found to effectively eliminate cancer cells and CSCs by blocking the Wnt/β-catenin and Notch signalling pathways in CRC.

Hedgehog-GLI and Notch Pathways Sustain Chemoresistance and Invasiveness in Colorectal Cancer and Their Inhibition Restores Chemotherapy Efficacy

“…Stem cells, transit amplifying cells and terminally differentiated secretory cells or enterocytes, concur in the formation of the structural unit of the colon, known as the crypt of Lieberkuhn [46]. A recent paper showed that the HH-GLI blockade with GANT61 was able to inhibit NOTCH and WNT/β-catenin in cellular models of CRC [47]. Since the HH-GLI and NOTCH pathways play a fundamental role in the correct patterning of the colonic mucosa and HH-GLI is upregulated by chemotherapeutic stress, we wondered whether HH-GLI and NOTCH crosstalk could be involved in the resistance mechanism of CRC cells related to 5-FU chemotherapeutic stress.…”