ABSTRACT
Chemotherapy resistance is a major challenge in ovarian cancer (OvCa). Thus, novel treatment combinations are highly warranted. However, many promising drug candidates tested in two-dimensional (2D) cell culture have not proved successful in the clinic. For this reason, we analyzed our drug combination not only in monolayers but also in three-dimensional (3D) tumor spheroids. One potential therapeutic target for OvCa is A disintegrin and metalloprotease 17 (ADAM17). ADAM17 can be activated by chemotherapeutics, which leads to enhanced tumor growth due to concomitant substrate cleavage. Therefore, blocking ADAM17 during chemotherapy may overcome resistance. Here, we tested the effect of the ADAM17 inhibitor GW280264X in combination with cisplatin on ovarian cancer cells in 2D and 3D. In 2D, the effect on five cell lines was analyzed with two readouts. Three of these cell lines formed dense aggregates or spheroids (HEY, SKOV-3, and OVCAR-8) in 3D and the treatment effect was analyzed with a multicontent readout (cytotoxicity, viability, and caspase3/7 activation). We tested the combined therapy on tumor spheroids derived from primary patient cells. In 2D, we found a significant reduction in the half minimal (50%) inhibitory concentration (IC50) value of the combined treatment (GW280264X plus cisplatin) in comparison with cisplatin monotherapy in all five cell lines with both 2D readout assays (viability and caspase activation). In contrast, the combined treatment only showed an IC50 reduction in HEY and OVCAR-8 3D tumor spheroid models using caspase3/7 activity or CelltoxTM Green as the readout. Finally, we found an improved effect of GW280264X with cisplatin in tumor spheroids derived from patient samples. In summary, we demonstrate that ADAM17 inhibition is a promising treatment strategy in ovarian cancer.
ABSTRACT
The spark plasma sintering (SPS) technique was employed to prepare compacts from (i) gas-atomized and (ii) attritor-milled AE42 magnesium powder. Short attritor-milling was used mainly to disrupt the MgO shell covering the powder particles and, in turn, to enhance consolidation during sintering. Compacts prepared by SPS from the milled powder featured finer microstructures than compacts consolidated from gas-atomized powder (i.e., without milling), regardless of the sintering temperatures in the range of 400-550 °C. Furthermore, the grain growth associated with the increase in the sintering temperature in these samples was less pronounced than in the samples prepared from gas-atomized particles. Consequently, the mechanical properties were significantly enhanced in the material made of milled powder. Apart from grain refinement, the improvements in mechanical performance were attributed to the synergic effect of the irregular shape of the milled particles and better consolidation due to effectively disrupted MgO shells, thus suppressing the crack formation and propagation during loading. These results suggest that relatively short milling of magnesium alloy powder can be effectively used to achieve superior mechanical properties during consolidation by SPS even at relatively low temperatures.
