Objective To explore the influence of Sinomenine（SIN）on the regulation of mitochondria-dependent cell apoptosis by modulating the sphingosine kinase 2（SPHK2）/myeloid cell leukemia 1 （MCL1） pathway in the context of acute lymphoblastic leukemia（ALL）progression.Methods ALL cell lines Jurkat were treated with different concentrations of SIN. Cell viability was measured using the CCK-8 assay，apoptosis was assessed using flow cytometry，intracellular ROS levels were detected using the DCFH-DA probe，and mitochondrial membrane potential（MMP）was evaluated using the JC-1 assay. Network pharmacology analysis was conducted to identify the targets of SIN in the treatment of ALL. An SPHK2 overexpression model was constructed to validate the mechanism of action of SIN in ALL. Additionally，a xenograft tumor model was established to assess the impact of SIN on the in vivo growth of Jurkat cells. Results Compared to the control group，treatment with SIN significantly suppressed ALL cell proliferation and induced apoptosis（P＜0.05）. The exposure of Jurkat cells to various concentrations of SIN resulted in an elevation of intracellular ROS levels and a reduction in MMP（P＜0.05）. Network pharmacology analysis identified SPHK2 as a key target of SIN in the treatment of ALL，which was further confirmed by experimental findings illustrating that SIN treatment inhibited the expression of both SPHK2 and MCL1（P＜0.05）. Moreover，overexpression of SPHK2 attenuated the anticancer effect of SIN in ALL（P＜0.05）. In vivo studies demonstrated that SIN treatment inhibited tumor growth and upregulated the expression of cleaved Caspase-3（P＜0.05）. Conclusion Sinomenine induces mitochondria-dependent apoptosis by inhibiting the SPHK2/MCL1 pathway，thereby suppressing the progression of ALL