B-cell acute lymphocytic leukemia (B-ALL) is a highly aggressive malignancy with poor prognosis. Developing diagnostic markers and therapeutic targets to identify and treat B-ALL early would improve the outcomes of B-ALL patients. Here, we conducted RNA next-generation sequencing using bone marrow (BM) specimens obtained from 7 B-ALL patients and 7 healthy donors. We found cysteine and glycine-rich protein 2 (CSRP2) upregulated in B-ALL. Down-regulation of CSRP2 resulted in suppressed cell proliferation and enhanced cell apoptosis in B-ALL. In addition, inhibition of CSRP2 increased cell ferroptosis in B-ALL cells. Mechanically, we revealed that transcription factor early B cell factor 1 (EBF1) regulated CSRP2 levels in B-ALL, and inhibition of EBF1 decreased CSRP2 levels in B-ALL. In conclusion, the dysregulation EBF1 led to CSRP2 upregulation and resulting in progression of B-ALL. The EBF1/CSRP2 axis could be of great potential as therapeutic targets for B-ALL treatment.