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BMC Complementary Medicine and Therapies
Published in: BMC Complementary Medicine and Therapies 1/2024

Open Access 01-12-2024 | Acute Lymphoblastic Leukemia | Research

Indigofera suffruticosa aerial parts extract induce G2/M arrest and ATR/CHK1 pathway in Jurkat cells

Authors: Hong-Loan Tran, Kuei-Hung Lai, Hsun-Shuo Chang, Yi-Siao Chen, Hui-Chun Wang, Shuen-Shin Yang, Hsueh-Wei Chang, Chin-Mu Hsu, Chia-Hung Yen, Hui-Hua Hsiao

Published in: BMC Complementary Medicine and Therapies | Issue 1/2024

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Abstract

Background

Indigofera suffruticosa Mill. is used as a folk medicine for treating patients with leukemia, however very little is known regarding the molecular mechanism of its anti-leukemic activity and the chemical profile of the active extract. The present study aimed to reveal the molecular effect of I. suffruticosa aerial parts extract (ISAE) on leukemia cells and its chemical constituents.

Methods

Cytotoxicity of ISAE were determined by resazurin viability assay, multitox – Glo multiplex cytotoxicity assay, and Annexin V staining assay. Cell cycle profiles were revealed by propidium iodide staining assay. The effects of ISAE on G2/M arrest signaling and DNA damage were evaluated by Western blot assay and phospho-H2A.X staining assay. The chemical profile of ISAE were determined by tandem mass spectroscopy and molecular networking approach.

Results

We showed that the acute lymphoblastic leukemia cell line Jurkat cell was more responsive to ISAE treatment than other leukemia cell lines. In contrast, ISAE did not induce cytotoxic effects in normal fibroblast cells. Cell cycle analysis revealed that ISAE triggered G2/M arrest in Jurkat cells in dose- and time-dependent manners. Elevation of annexin V-stained cells and caspase 3/7 activity suggested ISAE-induced apoptosis. Furthermore, ISAE alone could increase the phosphorylation of CDK1 at Y15 and activate the ATR/CHK1/Wee1/CDC25C signaling pathway. However, the addition of caffeine, a widely used ATR inhibitor to ISAE, reduced the phosphorylation of ATR, CHK1, and CDK1, as well as G2/M arrest in Jurkat cells. Moreover, increased phospho-H2A.X stained cells indicated the involvement of DNA damage in the anti-leukemic effect of ISAE. Finally, qualitative analysis using UPLC-tandem mass spectroscopy and molecular networking revealed that tryptanthrin was the most abundant organoheterocyclic metabolite in ISAE. At equivalent concentrations to ISAE, tryptanthrin induced G2/M arrest of Jurkat cells, which can be prevented by caffeine.

Conclusions

ISAE causes G2/M arrest via activating ATR/CHK1/CDK1 pathway and tryptanthrin is one of the active components of ISAE. Our findings provide subtle support to the traditional use of I. suffruitcosa in leukemia management in folk medicine.
Appendix
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Literature
1.
2.
Katz AJ, Chia VM, Schoonen WM, Kelsh MA. Acute lymphoblastic leukemia: an assessment of international incidence, survival, and disease burden. Cancer Causes Control. 2015;26(11):1627–42.PubMedCrossRef
3.
4.
Samra B, Jabbour E, Ravandi F, Kantarjian H, Short NJ. Evolving therapy of adult acute lymphoblastic leukemia: state-of-the-art treatment and future directions. J Hematol Oncol. 2020;13(1):70.PubMedPubMedCentralCrossRef
5.
Newman DJ, Cragg GM. Natural products as sources of new drugs over the nearly four decades from 01/1981 to 09/2019. J Nat Prod. 2020;83(3):770–803.PubMedCrossRef
6.
Williamson EM, Liu X, Izzo AA. Trends in use, pharmacology, and clinical applications of emerging herbal nutraceuticals. Br J Pharmacol. 2020;177(6):1227–40.PubMedPubMedCentralCrossRef
7.
Campos JKL, Araujo T, Brito T, da Silva APS, da Cunha RX, Martins MB, da Silva NH, Dos Santos BS, da Silva CA, Lima VLM. Indigofera suffruticosa Mill. (Anil): plant profile, phytochemistry, and pharmacology review. Adv Pharmacol Sci. 2018;2018:8168526.PubMedPubMedCentral
8.
Ou JC, Hsieh WC, Lin IH, Chang YS, Chen IS. The catalogue of medicinal plant resources in Taiwan. Taiwan: Department of Health, Executive Yuan, Taiwan; 2003.
9.
Yen CH, Lu YC, Li CH, Lee CM, Chen CY, Cheng MY, Huang SF, Chen KF, Cheng AL, Liao LY, et al. Functional characterization of glycine N-methyltransferase and its interactive protein DEPDC6/DEPTOR in hepatocellular carcinoma. Mol Med. 2012;18(1):286–96.PubMedCrossRef
10.
Liu W, Hsu YY, Tang JY, Cheng YB, Chuang YT, Jeng JH, Yen CH, Chang HW. Methanol extract of commelina plant inhibits oral cancer cell proliferation. Antioxidants. 2022;11(9):1813.PubMedPubMedCentralCrossRef
11.
Chen YS, Chang HS, Hsiao HH, Chen YF, Kuo YP, Yen FL, Yen CH. Identification of Beilschmiedia Tsangii root extract as a liver cancer cell-normal keratinocyte dual-selective NRF2 regulator. Antioxidants. 2021;10(4):544.PubMedPubMedCentralCrossRef
12.
Wang SH, Chen YS, Lai KH, Lu CK, Chang HS, Wu HC, Yen FL, Chen LY, Lee JC, Yen CH. Prinsepiae nux extract activates NRF2 activity and protects UVB-Induced damage in keratinocyte. Antioxidants. 2022;11(9):1755.PubMedPubMedCentralCrossRef
13.
14.
Tej G, Nayak PK. Mechanistic considerations in chemotherapeutic activity of caffeine. Biomed Pharmacother. 2018;105:312–9.PubMedCrossRef
15.
Blackford AN, Jackson SP. ATM, ATR, and DNA-PK: the trinity at the heart of the DNA damage response. Mol Cell. 2017;66(6):801–17.PubMedCrossRef
16.
Sedelnikova OA, Pilch DR, Redon C, Bonner WM. Histone H2AX in DNA damage and repair. Cancer Biol Ther. 2003;2(3):233–5.PubMedCrossRef
17.
18.
Molino R, Junio HA. Profiling the Philippine Blue: liquid chromatography/mass spectrometry-based metabolomics study on Philippine Indigofera. Rapid Commun Mass Spectrom. 2021;35(7):e9037.PubMedCrossRef
19.
Catanzaro E, Betari N, Arencibia JM, Montanari S, Sissi C, De Simone A, Vassura I, Santini A, Andrisano V, Tumiatti V, et al. Targeting topoisomerase II with trypthantrin derivatives: discovery of 7-((2-(dimethylamino)ethyl)amino)indolo[2,1-b]quinazoline-6,12-dione as an antiproliferative agent and to treat cancer. Eur J Med Chem. 2020;202:112504.PubMedCrossRef
20.
Song J, Kitamatsu M, Imamura K, Ohmori H, Watanabe K, Nakanishi K. On the preparation of indoxyl red from indican and some new characteristics. Bioorg Med Chem Lett. 2013;23(3):627–9.PubMedCrossRef
21.
Sun Q, Leng J, Tang L, Wang L, Fu C. A comprehensive review of the chemistry, pharmacokinetics, pharmacology, clinical applications, adverse events, and quality control of Indigo Naturalis. Front Pharmacol. 2021;12:664022.PubMedPubMedCentralCrossRef
22.
Tang W, Eisenbrand G. Qingdai. In: Tang W, Eisenbrand G, editors. Chinese drugs of plant origin. Berlin: Springer Berlin Heidelberg; 1992. p. 805–12.CrossRef
23.
Speranza J, Miceli N, Taviano MF, Ragusa S, Kwiecien I, Szopa A, Ekiert H. Isatis tinctoria L. (woad): a review of its botany, ethnobotanical uses, phytochemistry, biological activities, and biotechnological studies. Plants (Basel). 2020;9(3):298.PubMedCrossRef
24.
Yang L, Li X, Huang W, Rao X, Lai Y. Pharmacological properties of indirubin and its derivatives. Biomed Pharmacother. 2022;151:113112.PubMedCrossRef
25.
Wu X, Chen X, Dan J, Cao Y, Gao S, Guo Z, Zerbe P, Chai Y, Diao Y, Zhang L. Characterization of anti-leukemia components from indigo naturalis using comprehensive two-dimensional K562/cell membrane chromatography and in silico target identification. Sci Rep. 2016;6:25491.PubMedPubMedCentralCrossRef
26.
Hoessel R, Leclerc S, Endicott JA, Nobel ME, Lawrie A, Tunnah P, Leost M, Damiens E, Marie D, Marko D, et al. Indirubin, the active constituent of a Chinese antileukaemia medicine, inhibits cyclin-dependent kinases. Nat Cell Biol. 1999;1(1):60–7.PubMedCrossRef
27.
Eisenbrand G, Hippe F, Jakobs S, Muehlbeyer S. Molecular mechanisms of indirubin and its derivatives: novel anticancer molecules with their origin in traditional Chinese phytomedicine. J Cancer Res Clin Oncol. 2004;130(11):627–35.PubMedCrossRef
28.
Marko D, Schatzle S, Friedel A, Genzlinger A, Zankl H, Meijer L, Eisenbrand G. Inhibition of cyclin-dependent kinase 1 (CDK1) by indirubin derivatives in human tumour cells. Br J Cancer. 2001;84(2):283–9.PubMedPubMedCentralCrossRef
29.
Nam S, Scuto A, Yang F, Chen W, Park S, Yoo HS, Konig H, Bhatia R, Cheng X, Merz KH, et al. Indirubin derivatives induce apoptosis of chronic myelogenous Leukemia cells involving inhibition of Stat5 signaling. Mol Oncol. 2012;6(3):276–83.PubMedPubMedCentralCrossRef
30.
Chan HL, Yip HY, Mak NK, Leung KN. Modulatory effects and action mechanisms of tryptanthrin on murine myeloid leukemia cells. Cell Mol Immunol. 2009;6(5):335–42.PubMedPubMedCentralCrossRef
31.
Pathania AS, Kumar S, Guru SK, Bhushan S, Sharma PR, Aithagani SK, Singh PP, Vishwakarma RA, Kumar A, Malik F. The synthetic tryptanthrin analogue suppresses STAT3 signaling and induces caspase dependent apoptosis via ERK up regulation in human leukemia HL-60 cells. PLoS ONE. 2014;9(11):e110411.PubMedPubMedCentralCrossRef
32.
Gao JY, Chang CS, Lien JC, Chen TW, Hu JL, Weng JR. Synthetic tryptanthrin derivatives induce cell cycle arrest and apoptosis via akt and MAPKs in human hepatocellular carcinoma cells. Biomedicines. 2021;9(11):1527.PubMedPubMedCentralCrossRef
33.
Yang M, Tian X, Fan Z, Yu W, Li Z, Zhou J, Zhang W, Liang A. Targeting RAD51 enhances chemosensitivity of adult T–cell leukemia–lymphoma cells by reducing DNA double–strand break repair. Oncol Rep. 2019;42(6):2426–34.PubMedPubMedCentral
34.
Yu W, Li L, Wang G, Zhang W, Xu J, Liang A. KU70 inhibition impairs both non-homologous end joining and homologous recombination DNA damage repair through SHP-1 induced dephosphorylation of SIRT1 in T-cell acute lymphoblastic leukemia (T-ALL) [corrected]. Cell Physiol Biochem. 2018;49(6):2111–23.PubMedCrossRef
35.
Kimoto T, Hino K, Koya-Miyata S, Yamamoto Y, Takeuchi M, Nishizaki Y, Micallef MJ, Ushio S, Iwaki K, Ikeda M, et al. Cell differentiation and apoptosis of monocytic and promyelocytic leukemia cells (U-937 and HL-60) by tryptanthrin, an active ingredient of Polygonum Tinctorium Lour. Pathol Int. 2001;51(5):315–25.PubMedCrossRef
36.
Di Ghelli Luserna A, Ghetti M, Ledda L, Ferrari A, Bocconcelli M, Padella A, Napolitano R, Fontana MC, Liverani C, Imbrogno E, et al. Exploring the ATR-CHK1 pathway in the response of doxorubicin-induced DNA damages in acute lymphoblastic leukemia cells. Cell Biol Toxicol. 2023;39(3):795–811.CrossRef
37.
Hur J, Ghosh M, Kim TH, Park N, Pandey K, Cho YB, Hong SD, Katuwal NB, Kang M, An HJ, et al. Synergism of AZD6738, an ATR inhibitor, in combination with belotecan, a camptothecin analogue, in chemotherapy-resistant ovarian cancer. Int J Mol Sci. 2021;22(3):1223.PubMedPubMedCentralCrossRef
38.
Wengner AM, Siemeister G, Lucking U, Lefranc J, Wortmann L, Lienau P, Bader B, Bomer U, Moosmayer D, Eberspacher U, et al. The novel ATR inhibitor BAY 1895344 is efficacious as monotherapy and combined with DNA damage-inducing or repair-compromising therapies in preclinical cancer models. Mol Cancer Ther. 2020;19(1):26–38.PubMedCrossRef
39.
Jo U, Senatorov IS, Zimmermann A, Saha LK, Murai Y, Kim SH, Rajapakse VN, Elloumi F, Takahashi N, Schultz CW, et al. Novel and highly potent ATR inhibitor M4344 kills cancer cells with replication stress, and enhances the chemotherapeutic activity of widely used DNA damaging agents. Mol Cancer Ther. 2021;20(8):1431–41.PubMedPubMedCentralCrossRef
40.
Bradbury A, Hall S, Curtin N, Drew Y. Targeting ATR as cancer therapy: a new era for synthetic lethality and synergistic combinations? Pharmacol Ther. 2020;207:107450.PubMedCrossRef
41.
Kansagra A, Dahiya S, Litzow M. Continuing challenges and current issues in acute lymphoblastic leukemia. Leuk Lymphoma. 2018;59(3):526–41.PubMedCrossRef
42.
Samra B, Alotaibi AS, Short NJ, Khoury JD, Ravandi F, Garris R, Jain N, Konopleva M, Kantarjian H, Jabbour E. Outcome of adults with relapsed/refractory T-cell acute lymphoblastic leukemia or lymphoblastic lymphoma. Am J Hematol. 2020;95(9):E245-247.PubMedCrossRef
Metadata
Title
Indigofera suffruticosa aerial parts extract induce G2/M arrest and ATR/CHK1 pathway in Jurkat cells
Authors
Hong-Loan Tran
Kuei-Hung Lai
Hsun-Shuo Chang
Yi-Siao Chen
Hui-Chun Wang
Shuen-Shin Yang
Hsueh-Wei Chang
Chin-Mu Hsu
Chia-Hung Yen
Hui-Hua Hsiao
Publication date
01-12-2024
Publisher
BioMed Central
Published in
BMC Complementary Medicine and Therapies / Issue 1/2024
Electronic ISSN: 2662-7671
DOI
https://doi.org/10.1186/s12906-023-04325-w

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