Objective: To explore the pathogenic role of transcription factor zinc finger protein 326 (ZNF326) in T-cell acute lymphoblastic leukemia (T-ALL). Methods: The expression of ZNF326 in T-ALL tissues was analyzed using the TARGET database. Small interfering RNA (siRNA) technology was used to knock down the expression of the ZNF326 and serpin family B member 2 (SERPINB2) genes in Jurkat cells, and lentiviral transfection was performed to overexpress ZNF326 gene in Jurkat cells. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blot were used to verify the knockdown and overexpression efficiency. CCK-8 cell proliferation assay was performed to detect the proliferative capacity of cells after ZNF326 knockdown and overexpression, respectively, and to verify the effect of knocking down its downstream target gene SERPINB2 on cell proliferation. Flow cytometry was used to detect the effect of ZNF326 knockdown on apoptosis and cell cycle in T-ALL cells. RNA sequencing (RNA-seq) was performed to identify the downstream target genes of ZNF326 overexpression. Results: Knockdown of ZNF326 significantly inhibited the proliferation of T-ALL cells and induced G1 phase cell cycle arrest. Knockdown of ZNF326 downregulated the expression of SERPINB2, and specific knockdown of SERPINB2 also effectively inhibited the proliferation of T-ALL cells. Conclusion: ZNF326 may promote the proliferation of T-ALL cells by regulating the expression of SERPINB2, and may serve as a novel therapeutic target for T-ALL.