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Fig. 1 | Journal of Hematology & Oncology

Fig. 1

From: Cyclin D1-CDK4 activity drives sensitivity to bortezomib in mantle cell lymphoma by blocking autophagy-mediated proteolysis of NOXA

Fig. 1

Cyclin D1-driven CDK4 activity is required for cell death induction of NOXA accumulating agents. a CDK4 inhibition by palbociclib antagonizes bortezomib-induced cell death and NOXA accumulation. MCL cell lines Jeko-1 and Granta-519 were treated with 300 nM and Mino with 100 nM palbociclib for 16 h and subsequently co-treated with 8 nM bortezomib for 24 h. Primary MCL cells were exposed to 50 ng/ml interleukin-10, 50 ng/ml B cell activating factor, 1 ng/ml of insulin-like growth factor-1, 1 ng/ml interleukin-6, and co-cultured with the CD40 ligand-expressing cell line 3T3. After 8 h of co-culture, cells were treated with 300 nM palbociclib for 16 h and subsequently co-treated with 10 nM bortezomib. The 3T3 cell line was irradiated with 30 Gy 1 day before co-culture. After 8 h, protein expression was analyzed (right), and after 24 h, cell death was assessed by AnnexinV-PI staining (left). b Knockdown of cyclin D1 and CDK4 antagonizes bortezomib-induced cell death and NOXA accumulation. MCL cell line Mino was transfected with siRNA targeting CCND1 and CDK4. Twenty-four hours after transfection, cells were treated with 8 nM bortezomib. After 8 h, protein expression was analyzed (right) and cell death was assessed by AnnexinV-PI staining (left) 24 h post-treatment. c CDK4 inhibition antagonizes cell death of NOXA inducing substances and NOXA accumulation. MCL cell line Mino was treated with 100 nM palbociclib for 16 h and subsequently co-treated with either 8 nM carfilzomib, 20 μM orlistat, or 500 μM hydrogen peroxide. After 8 h, protein expression was analyzed (right), and after 24 h, treatment cell death was assessed by AnnexinV-PI staining (left). d MCL cell line Mino was transfected with siRNA targeting PMAIP1 and MCL1 and treated with 100 nM palbociclib. Twenty-four hours after transfection, protein expression was analyzed (right) and cells were treated with 8 nM bortezomib. Cell death was assessed by AnnexinV-PI staining 24 h post-treatment (left). Data represent means ± S.D. from three independent experiments

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