Fig. 6

Metformin and evodiamine cooperatively reduced stem cell phenotypes and tumor growth of PDAC. A Representative immunoblots of KLF10 and NICD-1, -3, and -4 in Panc-1 cells treated with various dosage of evodiamine (EVO) as indicated. β-Actin was used as internal control (left panel). Representative immunoblots of KLF10 and NICD-3 and -4 in Panc-1-shKLF10 treated with combination of 2 mM metformin (M) and/or 80 µM evodiamine (E) (right panel). B Sphere formation of Panc-1-pLKO (yellow) and Panc-1-pLKO-shKLF10 (blue) without treatment (filled) or treated with metformin alone (diagonal stripe) evodiamine alone (diagonal check) or with combination treatment (unfilled). The bar graphs were mean ± SE from cumulated data of three independent experiments. *p < 0.05. C Expression of CD24 (left panel) and c-MET (right panel) on Panc-1-pLKO (yellow) and Panc-1-pLKO-shKLF10 (blue) without treatment (filled), treated with metformin alone (diagonal stripe), with evodiamine alone (diagonal check), or with combination treatment (unfilled). The bar graphs were mean ± SE from cumulated data of three independent experiments. *p < 0.05 and **p < 0.001 respectively. D Schema of orthotopic murine model treated with metformin (120 mg/kg) and/or evodiamine (15 or 30 mg/kg). E (Left panel) representative IVIS images of mice bearing Panc-1-shKLF10 and treated with metformin (Met) and/or evodiamine (EVO). (Right panel) representative photos of tumors resected from murine experiments mentioned above. F Cumulated IVIS signal of at least six mice in each experimental group that were implanted with Panc-1-shKLF10 and received PBS (dashed yellow), metformin alone (solid yellow), evodiamine alone (dashed blue), or combination (solid blue) for 3 weeks, as indicated. Each point represents mean ± SE. *p < 0.05 and **p < 0.01. G Representative H&E (upper panel) and IHC staining, as described in “Materials and methods”, of KLF10, NICD-3 and NICD-4, as indicated, in tumors resected from mice experiments of E