Background: With the development of TKIs, the prognosis of Ph+ALL has been significantly improved. However, Ph+ALL especially with IKZF1 deletion are more likely to relapse and drug resistance. Huaier is a tranditional Chinese medicine, which has been shown an effective adjuvant of cancer therapy. .Our study aims to explore the effect and mechanism of Huaier on the chemosensitivity of IKZF1del-Ph+-ALL.
Methods: Cell viability was analysed by CCK8 and High-throughput Drug Sensitivity Analysis Strategy. Flow cytometry was used to detect cell apoptosis, cell cycle and adhesion markers. RNAseq, RT-PCR and Western blot were conducted to discover the related gene and protein expresson of stemness pathway.
Results: Our studies demonstrated that Huaier enhanced the efficacy of chemotherapy. Using the concentration of GI25, GI50 and GI75 of Huaier combined with VDLP, dasatinib, mercaptopurine and methotrexate, it was found that Huaier could improve the cell inhibition rate of these chemotherapy drugs. Huaier combined with dasatinib could significantly increased the apoptosis of Sup-B15 cells (Control group vs. dasatinib group vs. Huaier group vs. Huaier + dasatinib group: 7.70%±0.67% vs. 58.70%±0.35% vs. 60.97%±0.55% vs. 73.17%±0.44%, p < 0.0001), arrested the cell cycle in the G0/G1 phase (Control group vs. dasatinib group vs. Huaier group vs. Huaier + dasatinib group: 46.70%±0.05% vs. 53.74%±0.82% vs. 58.63%±0.01% vs. 63.70%±0.69%, p < 0.0001) and reduced the expression of adhesion-related molecules, such as the mRNA expression of ITGA4, ITGA8, THY-1, ITGA9 (p < 0.0001), ITGA5 (p = 0.0231), and the protein expression of FAK (p < 0.0001) and phosphorylated FAK (p = 0.0026).
The result of RNAseq showed that the differentially expressed genes were enriched in stemness-related pathways, among which Wnt/β-catenin and YAP were significantly down-regulated. The mRNA expression of wnt3, wnt3a, DVL-3, c-myc, FZ2, TCF-1, c-jun and cyclinD1 was significantly decreased (P < 0.0001). Also, the WNT pathways downstream of the key moleculars protein expression of β-catenin, c-myc, c-jun, TCF1, TCF3, TCF4 (P < 0.01) and corresponding phosphorylated proteins (P < 0.05) were all decreased. Also the mRNA (P < 0.0001) and protein expression of YAP, CTGF and survivin were significantly decreased (P < 0.01) after Huaier treatment.
Conclusions: In our study, we proved that Huaier can inhibit the Wnt/β-catenin and YAP pathway to enhance the chemotherapy sensitivity of Ph+ALL.Therefore, Huaier has great potential for clinical application in the treatment of refractory/relapsed Ph+ALL.
No relevant conflicts of interest to declare.
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