fig7

Figure 7. In vitro and in vivo validation of the synergistic anti-AML effects of Bortezomib and Venetoclax. (A-C) Dose-response curves and matrices for Venetoclax and Bortezomib in OCI-AML3 (ZIP synergy score: 19.905), U937 (ZIP synergy score: 14.365), and MOLM13/Vene (ZIP synergy score: 24.186) cells, with treatment for 24 h before viability measurement using CCK-8; (D-F) Apoptosis of the AML cell lines in Cluster 3 (U937, OCI-AML3, MOLM13/VENE) detected by flow cytometry after treatment with 6 nM Bortezomib, 3 μM Venetoclax, and a combination of Bortezomib and Venetoclax; (G) Apoptosis of primary cells from refractory AML patients detected by flow cytometry after treatment with 8 nM Bortezomib, 1 μM Venetoclax, and a combination of Bortezomib and Venetoclax; (H) Expression levels of apoptotic protein caspase-3 and its cleaved form in primary AML patient samples following treatment with Bortezomib, Venetoclax, and their combination; (I) Dose-response curves and matrices for Venetoclax and Bortezomib in stem cell from healthy donor (ZIP synergy score: -1.78); (J) Establishment of the Venetoclax-resistant mouse model and the corresponding dosing regimen; (K) Spleen morphology and weight in mice after combination treatment with Bortezomib (1 mg/kg) and Venetoclax (50 mg/kg); (L and M) Tumor burden (hCD45⁺ cells) in the bone marrow (L) and spleen (M) of mice following combination treatment with Bortezomib and Venetoclax. Results are expressed as mean ± SD of at least three independent experiments. Statistical significance was determined using an unpaired Student’s t-test. P < 0.05 was considered statistically significant. ns indicates not significant, * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001. AML: Acute myeloid leukemia; ZIP: zero interaction potency; CCK-8: Cell Counting Kit-8; SD: standard deviation; BTZ: Bortezomib; IC₅₀: half-maximal inhibitory concentration; APC: allophycocyanin; SSC: side scatter.