Oleanolic Acid Inhibits Malignant Progression of Leukemia K562 Cells Through The miR-18a-5p/STK4 Axis
Chronic myeloid leukemia(CML)is a disease caused by the malignant proliferation of hema-topoietic stem cells in the bone marrow.Although tyrosine kinase inhibitors(TKIs)have made significant progress in the treatment of chronic myeloid leukemia,the issue of drug resistance remains unresolved.Therefore,finding new therapeutic drugs and targets is crucial.Studies have shown that miRNAs are closely associated with various tumors,including chronic myeloid leukemia,and oleanolic acid(OA)has a good inhibitory effect on leukemia cells.Through transcriptome sequencing,it was found that oleanolic acid can significantly upregulate the level of serine/threonine protein kinase 4(STK4),and miRla-5p is the target miRNA of STK4.Therefore,this article aims to investigate whether oleanolic acid can affect the malignant progression of K562 cells through miR-18a-5p/STK4.The differentially expressed genes in K562 cells were enriched in apoptosis related pathways after treatment with oleanolic acid.The EdU ex-periment and CCK-8 experiment results showed that oleanolic acid reduced the proliferation ability of K562 cells(P＜0.05).qRT-PCR,immunofluorescence,and Western blot results showed that after treat-ment with oleanolic acid,the expression of STK4 protein was significantly upregulated(P＜0.05)and miR-18a-5p was downregulated in K562 cells(P＜0.05).Transfection of miR-18a-5p mimics into cells significantly inhibited the expression of STK4(P＜0.05);Transfection with miR-18a-5p inhibitor signifi-cantly increased the expression of STK4(P＜0.05).The results of reactive oxygen species(ROS)detec-tion and mitochondrial membrane potential detection showed that oleanolic acid can promote the increase of ROS in K562 cells and reduce mitochondrial membrane potential.After overexpression of STK4,the mitochondrial membrane potential of the cells decreased compared to the Vector group;Knocking down STK4 can reverse the decrease in mitochondrial membrane potential caused by the oleanolic acid group.Flow cytometry analysis showed that oleanolic acid can significantly promote the occurrence of cell apopto-sis,while the apoptosis rate significantly decreased after transfection with miR-18a-5p mimics(P＜0.05);Overexpression of STK4 can promote the transformation of cells from early apoptosis to late apop-tosis,while simultaneous transfection of miR-18a-5p mimics can inhibit this phenomenon.Our research results confirm that oleanolic acid can promote apoptosis of K562 cells by maintaining low expression of miR-18a-5p and high expression of STK4.
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