Fig. 8.

NSC-622608 overcomes TKI resistance via downregulation of the AKT/mTOR and JAK2/STAT5 pathways. (A) PCA results before batch removal for multiple datasets. Different colors represent different datasets. As shown in the schematic diagram, the three CML datasets (GSE130404, GSE44589, and GSE33224) (n=183) were separated without any intersection. PCA results after batch removal, as shown in the schematic diagram, showing the intersection of the three datasets, which can be used in subsequent analysis. There were 124 patients who achieved a drug response (IM-Rs), and 59 patients had no response (IM-NRs). The relative expression of STAT5 in the IM-NR group was greater than that in the IM-R group (p<0.001). (B) The correlations between the expression of VSIR and that of STAT5A and STAT5B were analyzed with Spearman’s correlation. The density curve on the right represents the trend in the distribution of STAT5A and STAT5B, and the upper density curve represents the trend in the distribution of VSIR expression. The abscissa and ordinate represent genes, and different colors represent different correlation coefficients (red represents a negative correlation); the darker the color is, the stronger the correlation. (C) Western blots showing decreased expression levels of the JAK2, STAT5, p-STAT5, AKT, p-AKT, p-mTOR, 4EBP1 and p70s6k proteins after treatment with NSC-622608 (0, 5, 10, 20 μM) in KBM5 and KBM5-T315I cells for 48 h. (D) At 48 h, the mitochondria-induced apoptosis pathway was increased in a concentration-dependent manner after treatment with NSC-622608 (0, 5, 10, or 20 μM) in KBM5 and KBM5-T315I cells. (E) qPCR revealed a decrease in JAK2 and STAT5 mRNA expression following VSIR knockdown. (F) The AKT/mTOR and JAK2/STAT5 pathways were more strongly activated in K562-R cells than in K562 cells according to Western blot analysis. Asterisks (*) indicate significance, *** indicates p<0.001, ** indicates p<0.01, * indicates p<0.05, and NS indicates no significance.