Objective To investigate the anti-cancer effect of Xanthatin on leukemia cells, and further to explore whether its mechanism is related to the regulation of long non-coding RNA(lncRNA) lysyl oxidase 1 antisense RNA1(LOXL1-AS1) and microRNA(MiR)-520d-5p expression. Methods Experimental groups: control group, Xanthatin 6 μmol/L group, Xanthatin 20 μmol/L group, Xanthatin 60 μmol/L group, si-NC group, si-LOXL1-AS1 group, and Xanthatin+pcDNA-LOXL1-AS1 group. Cell proliferation and apoptosis were detected by cell counting kit(CCK-8) method and flow cytometry. LOXL1-AS1 and miR-520d-5p expression was calculated using real-time quantitative PCR(RT-qPCR); Expression of B-cell lymphoma(Bcl)-2 and Bcl-related x protein(Bax) proteins were analyzed by Western blotting. The LOXL1-AS1 small interfering RNA and LOXL1-AS1 overexpression vector were respectively transfected into K-562 cells, and the effect of Xanthatin on cell proliferation, apoptosis, and expression of Bcl-2 and Bax proteins were detected using CCK-8, flow cytometry and Western blotting. The interaction between LOXL1-AS1 and miR-520d-5p was confirmed using dual luciferase reporter experiment. Results The cell proliferation inhibition rate, apoptosis rate, Bax protein expression, miR-520d-5p expression of K-562 cells were notably increased after Xanthatin treatment (P＜0.05), while Bcl-2 protein expression and LOXL1-AS1 expression were notably decreased(P＜0.05). After interference with LOXL1-AS1 expression, the cell proliferation inhibition rate, apoptosis rate, Bax protein expression, miR-520d-5p expression of K-562 cells were notably increased(P＜0.05), while Bcl-2 protein expression was notably decreased(P＜0.05). LOXL1-AS1 overexpression notably attenuated the effects of Xanthatin on K-562 cell proliferation, apoptosis and the expression of Bcl-2 and Bax proteins. LOXL1-AS1 directly bound to miR-520d-5p. Conclusion Xanthatin inhibits the proliferation and induce apoptosis of leukemia cells, the mechanism may be associated with the inhibition of LOXL1-AS1/miR-520d-5p axis.