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  • Synergistic Effects of Parthenolide with Vitamin C for improving of apoptosis by Anti-Apoptotic-targeting miR-181b/17/125b-5p on NALM-6 cells
  • Gholamhossein Tamaddon,1,* Atefeh Bahmei,2
    1. 1 Division of Hematology and Blood bank, Department of Medical Laboratory Sciences, School of Paramedical Sciences, Shiraz university of medical Sciences, Shiraz, Iran. 2 Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of
    2. Division of Hematology and Blood bank, Department of Medical Laboratory Sciences, School of Paramedical Sciences, Shiraz university of medical Sciences, Shiraz, Iran.


  • Introduction: Acute Lymphoblastic Leukemia (ALL) is one of the most common hematological malignancies in children. With currently available therapies, 70 % of patients can survive this disease with lower survival in adults over 65 years old. Parthenolide and vitamin C, natural compounds for cell death that can induce the apoptosis of cancer cells, have been explored as therapeutic agents for cancers. Also, it has been reported that miR-181b, miR-125b-5p, and miR-181b-5p are a dis-regulator of apoptosis, limiting the potential clinical use of cancer therapy. It is unclear whether parthenolide and vitamin C can have apoptotic activity on NALM-6 cells, or whether the combination of these two compounds can synergistically and affect these miRNAs.
  • Methods: We used MTT assay, and flow cytometry (single and dual staining) to detect the effects of parthenolide alone or in combination with vitamin C on NALM-6 cells. The levels of miR-181b/17/125b-5p were analyzed by real-time PCR. The mechanisms of miRNA-induced apoptosis by targeting PI3K were also predicted by bioinformatics study, Cytoscape, RNAhybrid, and Cell signaling enrichment. Molecular Docking was applied to validate that both parthenolide and vitamin C can affect these miRNAs.
  • Results: We observed that parthenolide and vitamin C alone or in combination can inhibit the cell viability of NALM-6 cells induce apoptosis and exert synergized effects. Both parthenolide and vitamin C, at all concentrations used, increased the number of early apoptotic and dead cells (late apoptotic) compared to the appropriate controls after 48 hours of incubation. An increase in the number of early apoptotic and dead (late-apoptotic) cells was observed after the co-incubation of parthenolide with vitamin C at concentrations of 1.925µM and 0.5 mM in comparison to each compound alone which ends in 82.52±1.520 % cell death (early and late-apoptotic cells) and only 6.50 ± 1.697% of cells survived. Moreover, we observed the rate of early apoptotic cells tended to be reduced when combination therapy was applied (only, 3.92%) We identified miR-17-5p, miR-125b-5p, and miR-181b-5p as targets for both parthenolide and vitamin C, experimentally and computationally. All three treatments - parthenolide, vitamin C, and the combination - decreased (downregulated) expression of the three miRNAs. For miR-17-5p, vitamin C caused a larger decrease in expression than parthenolide. The combination therapy reduced miR-17-5p expression more than parthenolide alone but less than vitamin C alone. A diverse pattern was seen for miR-125b-5p. Parthenolide lowered expression more than vitamin C, while the combination had a powerful effect. A similar pattern was seen for miR-181b-5p. They decreased this miRNA more than the two others and the combination therapy decreased this miRNA near to zero. Overall, parthenolide demonstrated the strongest inhibitory effects on the expression of miR-125b-5p and miR-181b-5p individually. The combination of parthenolide and vitamin C had the greatest suppressive effect on miR-181b-5p levels. On the other hand, parthenolide and vitamin C could inhibit cell viability and increase apoptosis by affecting the level of miR-17-5p, miR-125b-5p, and miR-181b-5p in NALM-6 cells, thus inhibiting the TGF-B and PI3K pathway and NF-KB signaling, and inducing apoptosis of Acute Lymphoblastic cells in vitro.
  • Conclusion: This current study suggests a theoretical basis for the combination of parthenolide and vitamin C for the treatment of B-ALL. Considering the safety of parthenolide and vitamin C in vivo, it is suggested that this combination therapy should be used in treatment, especially in tumors resistant to apoptosis. that parthenolide and vitamin C can be more explored as potential therapeutic approaches for ALL.
  • Keywords: miRNAs, combination therapy, parthenolide, vitamin C, B-ALL