Figure 9.

Activity of the Rarb/Runx3/Col6a1 signal axis is inhibited by metformin. Quantitative RT-PCR results show expression of Rarb, Runx3, and Col6a1 after metformin (20 mmol/L) and 2DG (10 mmol/L) treatment for 48 hours in hyperglycemic PDAC cells. (B) Western blot analysis shows expression of Rarb, Runx3, Col6a1, and phospho-AMPKα^Thr172 after metformin and 2DG treatment for 48 hours in hyperglycemic PDAC cells. (C) Quantitative RT-PCR analysis shows expression of Runx1, Runx2, Rara, Rarg, and Egr2 after metformin (20 mmol/L) and 2DG (10 mmol/L) treatment for 48 hours in hyperglycemic PDAC cells. (D) Glucose uptake assay shows reduced cellular uptake after metformin treatment for 24 hours. (E) Representative H&E pictures (left panel) and metastatic area quantification (right panel) show metastatic colonization in control and metformin treated hyperglycemic PDAC cells. All data are presented as mean ± standard deviation; data of 3 independent experiments are shown. Unpaired t test is applied, *P < .05. (F) Chromatin immunoprecipitation (ChIP) analysis of Rarb gene shows percentage input of H3K9me2, H3K27me3, and H3K27ac in control and metformin treated hyperglycemic PDAC cells. All data are presented as mean ± standard deviation, and data from 2 independent experiments are shown. H, hyperglycemic cells; Met, metformin.