UNASSIGNED: 270 Background: @entity4149 kinase inhibitors (TKI), such as @entity2240 , @entity2857 and @entity6219 , have replaced immunotherapy as the standard of care for metastatic @entity1358 (mRCC). However, their use in sequential or combined strategies is limited by the lack of evidences on the ability of TKIs to induce cell death in @entity5 cells. Aim of our study was to evaluate the different mechanisms responsible of the cytotoxic effects induced in vitro by   M doses of @entity2240 , @entity2857 and @entity6219 in 5637 and J82 @entity150 ( @entity150 ) cell lines. METHODS: The viability of @entity150 cell lines were tested by @entity98 assay. Autophagy was evaluated by western blot analysis with the anti-LC3 and anti- @entity10407 antibodies, @entity9186 staining and cytofluorimetric analysis. @entity39 and apoptosis, (    m) dissipation and ROS generation were determined by @entity4404 /PI, JC-1 and @entity21184 staining, respectively and cytofluorimetric analysis. The @entity467 activity was evaluated by ELISA. Finally, by mRNA estraction and RT-PCR array the @entity6219 -induced gene profile expression was evaluated. RESULTS: We found that treatment of 5637 and J82 @entity150 cells with the three TKI agents markedly reduced cell viability. Treatment for 24 h with @entity2240 and @entity2857 at 20   M dose, triggers an incomplete autophagy of @entity150 cells. In addition, inhibition of autophagy induced by @entity2240 and @entity2857 triggers cell death of @entity150 cells. Thus, @entity2240 by imparing the @entity467 activity induces lysosomal-dependent @entity39 . Similarly, @entity2857 by defective lysosomial degradation triggers ROS- and mitochondrial-dependent apoptosis. As regard to @entity6219 , we first demonstrate that treatment of @entity150 cells for 72 hrs (20   M) induces autophagic Type II cell death, which was markedly reversed in a dose-dependent manner by @entity8836 and @entity503 autophagic inhibitors. Finally, @entity6219 upregulates the mRNA expression of a-glucosidase (GAA) and @entity9486 belonging to the @entity876 @entity5 suppressor genes. CONCLUSIONS: Overall, our results showing different TKI-induced cell death mechanisms provide the rationale for the sequential use of these agents and the biological basis for novel molecularly targeted approaches.

Different effects of @entity2240 , @entity2857 , and XXXX on inducing @entity5 cell death: The role of autophagy.

@entity9486 :: ('7161', 'Gene') :: ['TP73'] @entity6219 :: ('MESH:C516667', 'Chemical') :: ['pazopanib'] @entity2240 :: ('MESH:C473478', 'Chemical') :: ['sunitinib'] @entity39 :: ('MESH:D009336', 'Disease') :: ['necrosis', 'Necrosis'] @entity5 :: ('MESH:D009369', 'Disease') :: ['tumor', 'cancer'] @entity4404 :: ('308', 'Gene') :: ['Annexin-V'] @entity467 :: ('1508', 'Gene') :: ['cathepsin B'] @entity2857 :: ('MESH:C471405', 'Chemical') :: ['sorafenib'] @entity10407 :: ('23636', 'Gene') :: ['p62'] @entity21184 :: ('MESH:C029569', 'Chemical') :: ['DCFDA'] @entity8836 :: ('C025946', 'Chemical') :: ['3MA'] @entity876 :: ('7157', 'Gene') :: ['p53'] @entity1358 :: ('MESH:D002292', 'Disease') :: ['renal cell carcinoma'] @entity150 :: ('MESH:D001749', 'Disease') :: ['BC', 'bladder cancer'] @entity98 :: ('CHEBI:53233', 'Chemical') :: ['MTT'] @entity9186 :: ('MESH:D000165', 'Chemical') :: ['acridine orange'] @entity503 :: ('MESH:D002738', 'Chemical') :: ['chloroquine'] @entity4149 :: ('D014443', 'Chemical') :: ['Tyrosine']
@entity6219 :: (MESH:C516667,Chemical) :: ['pazopanib']
