Oridonin Could Inhibit Inflammation and T-cell Immunoglobulin and Mucin-3/Galectin-9 (TIM-3/Gal-9) Autocrine Loop in the Acute Myeloid Leukemia Cell Line (U937) as Compared to Doxorubicin
The T-cell immunoglobulin and mucin-3 (TIM-3)/galectin-9 (Gal-9) autocrine loop is an indispensable signaling in acute myeloid leukemia (AML) cells, which induces their self-renewal through activation of nuclear factor-kappa b (NF-kB) and β-catenin pathways. In this study, we evaluated the effects of oridonin and doxorubicin on the TIM-3/Gal-9 autocrine loop. We also evaluated oridonin anti-inflammatory and anti-cancer properties on U937 cells, as an AML cell line in comparison to doxorubicin as a common anthracycline drug for AML treatment. Cell counting kit-8 (CCK-8) was applied to evaluate the cytotoxicity of oridonin and doxorubicin on U937 cells and also to determine the impact of galectin-9 (Gal-9) on their proliferation. The effects of oridonin and doxorubicin on Gal-9, TIM-3, and interleukin-1β (IL-1β) gene expression were determined by real-time polymerase chain reaction (RT-PCR). The Gal-9 secretion level was measured by enzyme-linked immunosorbent assay (ELISA) and activation of NF-kB pathway was assessed by western blotting. In a dose-dependent manner, oridonin and doxorubicin were capable to eradicate U937 cells while Gal-9 expanded them. Following the treatment of U937 cells with oridonin, the expression of Gal-9, TIM-3, and IL-1β genes was down-regulated, and the Gal-9 secretion and NF-kB phosphorylation were diminished, whereas doxorubicin increased all of these factors. Doxorubicin is a common treatment agent in AML, but it may induce inflammation and up-regulate the TIM3/Gal-9 autocrine loop, consequently can enhance the possibility of disease relapse. Meanwhile, oridonin is capable to inhibit the essential signaling pathways in AML cells and reduce the inflammation and expansion of tumor cells and postpone AML recurrence.
2. Jones LM, Melgar K, Bolanos L, Hueneman K, Walker MM, Jiang J-K, et al. Targeting AML-associated FLT3 mutations with a type I kinase inhibitor. J Clin Invest. 2020;130(4).
3. De Kouchkovsky I, Abdul-Hay M. Acute myeloid leukemia: a comprehensive review and 2016 update. Blood Cancer J. 2016;6(7):e441–e441.
4. Naito K, Ohnishi K. [Current and new therapeutic strategies in acute myeloid leukemia]. Gan To Kagaku Ryoho. 2005;32(3):292–6.
5. Guo R, Wu K, Chen J, Mo L, Hua X, Zheng D, et al. Exogenous Hydrogen Sulfide Protects against Doxorubicin-Induced Inflammation and Cytotoxicity by Inhibiting p38MAPK/NFκB Pathway in H9c2 Cardiac Cells. Cell Physiol Biochem. 2013;32(6):1668–80.
6. Ibrahim A, Al-Hizab FA, Abushouk AI, Abdel-Daim MM. Nephroprotective Effects of Benzyl Isothiocyanate and Resveratrol Against Cisplatin-Induced Oxidative Stress and Inflammation. Front Pharmacol. 2018;9.
7. Debatin K-M. Apoptosis pathways in cancer and cancer therapy. Cancer Immunol Immunother. 2004;53(3):153–9.
8. Wang CY, Guttridge DC, Mayo MW, Baldwin AS. NF-kappaB induces expression of the Bcl-2 homologue A1/Bfl-1 to preferentially suppress chemotherapy-induced apoptosis. Mol Cell Biol. 1999;19(9):5923–9.
9. Chen R, Alvero AB, Silasi D-A, Mor G. Inflammation, Cancer and Chemoresistance: Taking Advantage of the Toll-Like Receptor Signaling Pathway. Am J Reprod Immunol. 2007;57(2):93–107.
10. Kikushige Y, Shima T, Takayanagi SI, Urata S, Miyamoto T, Iwasaki H, et al. A TIM-3/Gal-9 Autocrine Stimulatory Loop Drives Self-Renewal of Human Myeloid Leukemia Stem Cells and Leukemic Progression. Cell Stem Cell. 2010 Dec;7(6):708–17.
11. Progression L, Kikushige Y, Miyamoto T, Yuda J, Jabbarzadeh-tabrizi S, Shima T. A TIM-3 / Gal-9 Autocrine Stimulatory Loop Drives Self-Renewal of Human Myeloid Leukemia Stem Article A TIM-3 / Gal-9 Autocrine Stimulatory Loop Drives Self-Renewal of Human Myeloid Leukemia Stem Cells and Leukemic Progression. Stem Cell. 2015;1–12.
12. Thorn CF, Oshiro C, Marsh S, Hernandez-Boussard T, McLeod H, Klein TE, et al. Doxorubicin pathways: Pharmacodynamics and adverse effects. Pharmacogenet Genomics. 2011;21(7):440–6.
13. Sauter KAD, Wood LJ, Wong J, Iordanov M, Magun BE. Doxorubicin and daunorubicin induce processing and release of interleukin-1β through activation of the NLRP3 inflammasome: Progress at a snail’s pace. Cancer Biol Ther. 2011;11(12):1008–16.
14. Wu J, Ding Y, Chen C-H, Zhou Z, Ding C, Chen H, et al. Wu, J., et al., A new oridonin analog suppresses triple-negative breast cancer cells and tumor growth via the induction of death receptor 5. Cancer letters, 2016. 380(2):393-402.
15. Wang S-Q, Wang C, Chang L-M, Zhou K-R, Wang J-W, Ke Y, et al. Geridonin and paclitaxel act synergistically to inhibit the proliferation of gastric cancer cells through ROS-mediated regulation of the PTEN/PI3K/Akt pathway. Oncotarget. 2016;7(45):72990–3002.
16. Shang C, Zhang Q, Zhou J. Oridonin Inhibits Cell Proliferation and Induces Apoptosis in Rheumatoid Arthritis Fibroblast-Like Synoviocytes. Inflammation. 2016;39(2):873–80.
17. Leung CH, Grill SP, Lam W, Han Q Bin, Sun HD, Cheng YC. Novel mechanism of inhibition of nuclear factor-κB DNA-binding activity by diterpenoids isolated from Isodon rubescens. Mol Pharmacol. 2005;68(2):286–97.
18. Yi S, Chen Y, Wen L, Yang L, Cui G. Downregulation of nucleoporin 88 and 214 induced by oridonin may protect OCIM2 acute erythroleukemia cells from apoptosis through regulation of nucleocytoplasmic transport of NF-κB. Int J Mol Med. 2012;30(4):877–83.
19. Döhner H, Weisdorf DJ, Bloomfield CD. Acute myeloid leukemia. N Engl J Med. 2015;373(12):1136–52.
20. Chung HY, Kim DH, Lee EK, Chung KW, Chung S, Lee B, et al. Redefining chronic inflammation in aging and age-related diseases: Proposal of the senoinflammation concept. Vol. 10, Aging and Disease. International Society on Aging and Disease; 2019.8(14):367–82.
21. Liu T, Zhang L, Joo D, Sun SC. NF-κB signaling in inflammation. Signal Transduct Target Ther. 2017;2.
22. Park MH, Hong JT. Roles of NF-κB in Cancer and Inflammatory Diseases and Their Therapeutic Approaches. Cells. 2016;5(2).
23. Lewis AM, Varghese S, Xu H, Alexander HR. Interleukin-1 and cancer progression : the emerging role of interleukin-1 receptor antagonist as a novel therapeutic agent in cancer treatment. 2006;12:1–12.
24. Voronov E, Carmi Y, Apte RN. The role IL-1 in tumor-mediated angiogenesis. 2014;5:1–12.
25. Carey A, V DKE, Eide CA, Bagby GC, Mcweeney SK, Carey A, et al. Identification of Interleukin-1 by Functional Screening as a Key Mediator of Cellular Expansion and Disease Progression in Acute Myeloid Leukemia. Cell Reports. 2017;18(13):3204–18.
26. Sakhnevych SS, Yasinska IM, Bratt AM, Benlaouer O, Silva IG, Hussain R, et al. Cortisol facilitates the immune escape of human acute myeloid leukemia cells by inducing latrophilin 1 expression. Cell Mol Immunol. 2018;4–7.
27. Gonçalves Silva I, Yasinska IM, Sakhnevych SS, Fiedler W, Wellbrock J, Bardelli M, et al. The Tim-3-galectin-9 Secretory Pathway is Involved in the Immune Escape of Human Acute Myeloid Leukemia Cells. EBioMedicine. 2017;22:44–57.
28. Nagahara K, Arikawa T, Oomizu S, Kontani K, Nobumoto A, Tateno H, et al.Galectin-9 Increases Tim-3+ Dendritic Cells and CD8+ T Cells and Enhances Antitumor Immunity via Galectin-9-Tim-3 Interactions . J Immunol. 2008;181(11):7660–9.
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