Figure 7

The combination treatment of MMC and proteasomal inhibitor reduces tumor progression in HeLa cells xenografted mouse tumor model. (a) Experimental layout of in vivo study. HeLa cells (1 × 10⁶ in 100 μl PBS) were injected on the right flank of the mice to form tumors. Tumor-bearing mice were treated with combination of MMC (1 mg/kg/every third day) and MG132 (10 μM/kg/day) as described in Materials and Methods. Control mice were administered with equal volume of vehicle on the same treatment day. (b) Tumor progression after drug administration in control and treated mice. (c and d) Bar graph showing tumor volume and tumor weight in mice at the end of the experiment. (e) Changes in body weight in mice during the course of the experiment. (f) Histopathological analysis of major vital organs collected from experimental mice. Kidney, heart, liver and lungs were fixed in 4% formaldehyde. The tissues sections were stained with hematoxylin and eosin (H&E; magnification, × 400). (g) TAMs were isolated from tumor as described in Materials and Methods. Cells were analyzed for FasL expression by flow cytometry. Cells were dually stained with CD11b (1 : 100) and FasL (1 : 100), and CD11b-positive cells were gated to analyze FasL expression. (h) Representative images of immunostained section analysis of Fas (i) TUNEL assay (ii) in tumor tissues of different treatment groups (magnification, × 40 with inset at × 400)