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. 2016 Dec 27;8(12):19403–19412. doi: 10.18632/oncotarget.14292

Figure 3. Heparanase overexpression enhances insulin receptor signaling pathway and augments proliferative response to insulin in MCF-7 cells.

Figure 3

A. Heparanase overexpression in MCF7-Hpa (as compared to control MCF7-mock) cells was confirmed by activity assay, as described in Method and refs.(19, 54). B. MCF7-mock and MCF7-Hpa cells were serum-starved overnight and then either remained untreated (c) or stimulated with insulin (100 nM) for 15 and 30 min. Cell lysates containing equivalent amounts of total protein were then immunoblotted using antibody specific for pINSR, pAKT, total INSR, total AKT or total actin. The data shown are representative of three independent experiments. C. Presence of heparanase specific inhibitor SST0001 (10 μg/ml) reduced pINSR levels in MCF7-Hpa cells treated with insulin. D. Bar graph demonstrates the increase in proliferation of MCF7-mock and MCF7-Hpa cells cultured for 72 h in the absence (gray bars) or presence of insulin (100 nM, black bars), analyzed by MTS Cell Proliferation Assay. Note that presence of insulin does not confer statistically significant increase in proliferation of MCF7-mock cells (lacking heparanase activity). In contrast, in heparanase overexpressing MCF7-Hpa cells proliferation rate was significantly higher in the presence of insulin. *p<0.04 Two-sided Student's t test. Error bars represent ± SD. The data shown are representative of three independent experiments.