Research Paper Volume 11, Issue 10 pp 3376—3391

Figure 5. MiR-335 contributed to leukemogenesis through PI3K/Akt signaling pathway. (A) Expression heatmap of top 50 differentially expressed genes (DEGs) between ID4^low and ID4^high AML patients among TCGA datasets (FDR<0.05, P<0.05 and |log2 FC|>1.5). (B) Volcano plot of DEGs. (C) Significantly enriched pathway terms of DEGs in AML. DEGs functional and signaling pathway enrichment was conducted using online website of STRING (http://string-db.org). (D) GSEA showed the significantly enriched in genes down-regulated in mouse prostate by transgenic expression of human AKT1 gene in ID4^high AML. (E) The association between ID4 expression and AKT1/AKT2 expression in AML among TCGA datasets. A negative correlation was observed between ID4 and AKT1/AKT2 expression. The correlation analysis conducted through online web GEPIA (http://gepia.cancer-pku.cn/detail.php?clicktag=correlation) using Spearman test. (F) The expression of Akt and pAkt affected by miR-335 overexpression and ID4 restoration. Akt and pAkt protein expression was significantly increased after miR-335 overexpression in both K562 and HL60 cell-lines, and was decreased after ID4 restoration. (G–H) The effect of miR-335 on proliferation of K562 and HL60 cell-lines with/without Akt inhibitor MK2206 2HCL. MK2206 2HCL remarkably reversed and impaired the pro-proliferative effect in K562 and HL60 cell-lines. (I–J) The effect of miR-335 on apoptosis of K562 and HL60 cell-lines with/without Akt inhibitor MK2206 2HCL. MK2206 2HCL remarkably reversed and impaired the pro-proliferative effect in K562 and HL60 cell-lines. *, P<0.05; **, P<0.01; ***, P<0.001.