Isatin-mediated inhibition of the PI3K/AKT/EMT signaling axis confers antitumor efficacy in Ehrlich solid carcinoma-bearing mice

Despite advances, detailed insight into cancer’s biological mechanisms remains needed. Isatin, an indole-1 H-2,3-dione, is an attractive agent for new drug discovery. Here, the anticancer efficacy and potential toxicity of isatin in combination with doxorubicin (Dox) in a mouse Ehrlich solid carcinoma (ESC) model were examined by modulating the PI3K/AKT/EMT pathway. Five groups of fifty female mice were prepared: negative control, positive ESC-bearing mice, isatin-treated ESC group, Dox-treated ESC group, and isatin and Dox combination-treated ESC group. Treatments began after tumor formation. Inflammatory markers, kidney and liver enzyme activities, and EMT markers levels (vimentin and E-cadherin) were assessed by ELISA. HIF 1 α, NF-κB, PI3K, AKT, MMP2, and MMP9 gene expression was evaluated using qRT-PCR. Protein expression of MMP-9 and E-cadherin was assessed by IHC. Compared with the positive control, the combined treatment reduced tumor growth by 51%. Additionally, combined treatment showed a significant increase in the amount of necrotic tissue. Mechanistically, isatin exerted its therapeutic effect by increasing anti-inflammatory (IL-10) cytokine levels and suppressing pro-inflammatory (IL-6) cytokine levels, along with reductions in HIF-1α, NF-κB, PI3K, AKT, MMP2, and MMP9 mRNA levels. Also, a combined group of isatin and Dox showed reduced vimentin levels and elevated E-cadherin levels by ELISA. Co-administration of isatin and Dox significantly increased E-cadherin expression and attenuated MMP-9 expression compared with the untreated group, as assessed by IHC. Our results indicate that isatin, in combination with doxorubicin, exerts a synergistic anticancer effect against ESC, offering a promising therapeutic approach to enhance treatment efficacy.