Figure 1.
MIF is highly expressed in gastric and colon tumors and human tissue-derived fibroblasts.
MIF mRNA levels measured by qRT PCR are increased in A) human gastric tumor samples and at higher levels in tissues from patients with nodal involvement, B) human colon tumor samples and at higher levels in tissues from patients with nodal involvement, and C) in the supernatants tumor derived gastric and colon fibroblasts compared to matched normal as measured by Luminex singleplex assay. N = 8 for C and the mean ± standard error are shown as the results of duplicated in multiple experiments. *p<0.05.
Figure 2.
CD74 is highly expressed in human gastric and colon tumors and on isolated epithelial cells.
CD74 mRNA levels measured by qRT PCR are increased in A) human gastric tumor samples and at higher levels in tissues from patients with nodal involvement, B) human colon tumor samples and at higher levels in tissues from patients with nodal involvement, and on epithelial cells isolated from human gastric and colon tumors in C) a representative flow cytometry plot and D) in compiled flow cytometry data from all samples. N = 8 for D and the mean ± standard error are shown as the results of duplicated in multiple experiments. *p<0.05.
Figure 3.
MIF induces proliferation of gastric and colon carcinoma cells.
Tumor-derived gastric and colon supernatants induced proliferation of N87 and Caco-2 cells, which was decreased upon adding A) anti-MIF neutralizing antibodies or anti-CD74 blocking antibodies. B) Chronic exposure of HS738 or N87 cells with recombinant MIF increased proliferation that was sustained after returning cells to regular media for 8 weeks. N = 8 and the mean ± standard error are shown as the results of duplicated in multiple experiments. *p<0.05.
Figure 4.
MIF induces pro-tumorigenic signaling.
Chronic MIF treatment induces A) Akt, B) c-Jun, and C) Erk1/2 phosphorylation. N = 8 and the mean ± standard error are shown as the results of duplicated in multiple experiments. *p<0.05.
Figure 5.
Chronic MIF exposure induces mesenchymal epithelial transition.
HS738 cells have A) a fibroblast morphology, which changes B) upon chronic exposure to recombinant MIF. MIF exposure also decreases expression of fibroblast markers by C) mRNA levels by qRT PCR compared to untreated cells and by D) flow cytometry, while increasing expression of epithelial markers EpCam and E-cadherin by E) mRNA levels by qRT PCR compared to untreated cells and by D) flow cytometry. N = 8 and the mean ± standard error are shown as the results of duplicated in multiple experiments. *p<0.05.
Figure 6.
Chronic MIF exposure induces HS738 cell transformation.
HS738 chronic exposure to MIF induces A) upregulation of TERT gene expression so similar levels as N87 control cells and B) colony formation in a focus forming assay. N = 8 and the mean ± standard error are shown as the results of duplicated in multiple experiments. *p<0.05.