Figure 5 The effects of flow-induced shear stress on 3D ovarian cancer biology. (A) Western blot analysis of OVCAR-5 tumors was performed 7 days after culture under static or flow conditions. A flow-induced increase in EGFR and p-ERK, compared to static cultures, was observed. Conversely, a reduction in p-FAK, p-Paxillin, and Vinculin was observed under flow, relative to static conditions. (B) Western blot analysis of 3D OVCAR-5 tumors was performed 11 days after culture under static or flow conditions, including 4 days of treatment with 500 µM carboplatin, and respective controls. In both static and flow 3D cultures, carboplatin treatment resulted in downregulation of EGFR, FAK, p-Paxillin, Paxillin, and Vinculin. Upregulation of p-ERK was observed after carboplatin treatment in both static and flow 3D cultures. (C) Baseline levels of EGFR activity and expression are maintained by a complex array of factors, including recycling and degradation of the activated receptor complex. Flow-induced shear stress has been shown to cause a posttranslational up-regulation of EGFR expression and activation, likely resulting from increased receptor recycling and decreased EGFR degradation. Activation of EGFR results in ERK phosphorylation to induce gene expression, ultimately leading to cell proliferation, survival, and chemoresistance. FAK and other tyrosine kinases are activated by the engagement of integrins with the ECM. Subsequent phosphorylation of paxillin by FAK not only influences the remodeling of the actin cytoskeleton, but also modulates vinculin activation to regulate mitogen-activated protein kinase (MAPK) cascades, thereby stimulating pro-survival gene expression.

Figure 5
The effects of flow-induced shear stress on 3D ovarian cancer biology. (A) Western blot analysis of OVCAR-5 tumors was performed 7 days after culture under static or flow conditions. A flow-induced increase in EGFR and p-ERK, compared to static cultures, was observed. Conversely, a reduction in p-FAK, p-Paxillin, and Vinculin was observed under flow, relative to static conditions. (B) Western blot analysis of 3D OVCAR-5 tumors was performed 11 days after culture under static or flow conditions, including 4 days of treatment with 500 µM carboplatin, and respective controls. In both static and flow 3D cultures, carboplatin treatment resulted in downregulation of EGFR, FAK, p-Paxillin, Paxillin, and Vinculin. Upregulation of p-ERK was observed after carboplatin treatment in both static and flow 3D cultures. (C) Baseline levels of EGFR activity and expression are maintained by a complex array of factors, including recycling and degradation of the activated receptor complex. Flow-induced shear stress has been shown to cause a posttranslational up-regulation of EGFR expression and activation, likely resulting from increased receptor recycling and decreased EGFR degradation. Activation of EGFR results in ERK phosphorylation to induce gene expression, ultimately leading to cell proliferation, survival, and chemoresistance. FAK and other tyrosine kinases are activated by the engagement of integrins with the ECM. Subsequent phosphorylation of paxillin by FAK not only influences the remodeling of the actin cytoskeleton, but also modulates vinculin activation to regulate mitogen-activated protein kinase (MAPK) cascades, thereby stimulating pro-survival gene expression.