Top

BMC Complementary Medicine and Therapies

Published in:

Open Access 01-12-2021 | Breast Cancer | Research article

Thymoquinone inhibited vasculogenic capacity and promoted mesenchymal-epithelial transition of human breast cancer stem cells

Authors: Sanya Haiaty, Mohammad-Reza Rashidi, Maryam Akbarzadeh, Ahad Bazmani, Mostafa Mostafazadeh, Saba Nikanfar, Zohre Zibaei, Reza Rahbarghazi, Mohammad Nouri

Published in: BMC Complementary Medicine and Therapies | Issue 1/2021

Abstract

Background

Vasculogenic mimicry (VM) is characterized by the formation of tubular structure inside the tumor stroma. It has been shown that a small fraction of cancer cells, namely cancer stem cells (CSCs), could stimulate the development of vascular units in the tumor niche, leading to enhanced metastasis to the remote sites. This study aimed to study the inhibitory effect of phytocompound, Thymoquinone (TQ), on human breast MDA-MB-231 cell line via monitoring Wnt/PI3K signaling pathway.

Methods

MDA-MB-231 CSCs were incubated with different concentrations of TQ for 48 h. The viability of CSCs was determined using the MTT assay. The combination of TQ and PI3K and Wnt3a inhibitors was examined in CSCs. By using the Matrigel assay, we measured the tubulogenesis capacity. The percent of CD24⁻ CSCs and Rhodamine 123 efflux capacity was studied using flow cytometry analysis. Protein levels of Akt, p-Akt, Wnt3a, vascular endothelial-cadherin (VE-cadherin), and matrix metalloproteinases-2 and -9 (MMP-2 and -9) were detected by western blotting.

Results

TQ decreased the viability of CSCs in a dose-dependent manner. The combination of TQ with PI3K and Wnt3a inhibitors reduced significantly the survival rate compared to the control group (p < 0.05). TQ could blunt the stimulatory effect of vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), fibroblast growth factor (FGF) on CSCs (p < 0.05). The vasculogenic capacity of CSCs was reduced after being-exposed to TQ (p < 0.05). Western blotting revealed the decrease of CSCs metastasis by suppressing MMP-2 and -9. The protein level of VE-cadherin was also diminished in TQ-treated CSCs as compared to the control cell (p < 0.05), indicating inhibition of mesenchymal-endothelial transition (MendT). TQ could suppress Wnt3a and PI3K, which coincided with the reduction of the p-Akt/Akt ratio. TQ had the potential to decrease the number of CD24⁻ CSCs and Rhodamine 123 efflux capacity after 48 h.

Conclusion

TQ could alter the vasculogenic capacity and mesenchymal-epithelial transition of human breast CSCs in vitro. Thus TQ together with anti-angiogenic therapies may be a novel therapeutic agent in the suppression of VM in breast cancer.

Zamani ARN, Mashayekhi MR, Jadid MFS, Faridvand Y, Tajalli H, Rahbarghazi R. Photo-modulation of zinc phthalocyanine-treated breast cancer cell line ZR-75-1 inhibited the normal tumor activity in vitro. Lasers Med Sci. 2018;33(9):1969–78 https://doi.org/10.1007/s10103-018-2563-0.PubMedCrossRef

Maroufi NF, Amiri M, Dizaji BF, Vahedian V, Akbarzadeh M, Roshanravan N, Haiaty S, Nouri M, Rashidi MR. Inhibitory effect of melatonin on hypoxia-induced vasculogenic mimicry via suppressing epithelial-mesenchymal transition (EMT) in breast cancer stem cells. Eur J Pharmacol. 2020;881:173282 https://doi.org/10.1016/j.ejphar.2020.173282.PubMedCrossRef

Qi L, Song W, Liu Z, Zhao X, Cao W, Sun B. Wnt3a promotes the Vasculogenic mimicry formation of Colon Cancer via Wnt/beta-catenin signaling. Int J Mol Sci. 2015;16(8):18564–79 https://doi.org/10.3390/ijms160818564.PubMedPubMedCentralCrossRef

Cheraghi O, Dehghan G, Mahdavi M, Rahbarghazi R, Rezabakhsh A, Charoudeh HN, Iranshahi M, Montazersaheb S. Potent anti-angiogenic and cytotoxic effect of conferone on human colorectal adenocarcinoma HT-29 cells. Phytomedicine. 2016;23(4):398–405 https://doi.org/10.1016/j.phymed.2016.01.015.PubMedCrossRef

Haiaty S, Rashidi MR, Akbarzadeh M, Maroufi NF, Yousefi B, Nouri M. Targeting vasculogenic mimicry by phytochemicals: a potential opportunity for cancer therapy. IUBMB Life. 2020;72(5):825–41 https://doi.org/10.1002/iub.2233.PubMedCrossRef

Rahbarghazi R, Nassiri SM, Khazraiinia P, Kajbafzadeh AM, Ahmadi SH, Mohammadi E, Molazem M, Zamani-Ahmadmahmudi M. Juxtacrine and paracrine interactions of rat marrow-derived mesenchymal stem cells, muscle-derived satellite cells, and neonatal cardiomyocytes with endothelial cells in angiogenesis dynamics. Stem Cells Dev. 2013;22(6):855–65 https://doi.org/10.1089/scd.2012.0377.PubMedCrossRef

Amini H, Rezaie J, Vosoughi A, Rahbarghazi R, Nouri M. Cardiac progenitor cells application in cardiovascular disease. J Cardiovasc Thorac Res. 2017;9(3):127–32.PubMedPubMedCentralCrossRef

Nemati S, Rezabakhsh A, Khoshfetrat AB, Nourazarian A, Avci ÇB, Bagca BG, Sardroud HA, Khaksar M, Ahmadi M, Delkhosh A, Sokullu E, Rahbarghazi R. Alginate-gelatin encapsulation of human endothelial cells promoted angiogenesis in in vivo and in vitro milieu. Biotechnol Bioeng. 2017;114(12):2920–30.PubMedCrossRef

Cao Z, Bao M, Miele L, Sarkar FH, Wang Z, Zhou Q. Tumour vasculogenic mimicry is associated with poor prognosis of human cancer patients: a systemic review and meta-analysis. Eur J Cancer. 2013;49(18):3914–23 https://doi.org/10.1016/j.ejca.2013.07.148.PubMedCrossRef

10.

Sun B, Zhang S, Zhao X, Zhang W, Hao X. Vasculogenic mimicry is associated with poor survival in patients with mesothelial sarcomas and alveolar rhabdomyosarcomas. Int J Oncol. 2004;25(6):1609–14.PubMed

11.

Shang B, Cao Z, Zhou Q. Progress in tumor vascular normalization for anticancer therapy: challenges and perspectives. Front Med. 2012;6(1):67–78 https://doi.org/10.1007/s11684-012-0176-8.PubMedCrossRef

12.

Dianat-Moghadam H, Heidarifard M, Jahanban-Esfahlan R, Panahi Y, Hamishehkar H, Pouremamali F, Rahbarghazi R, Nouri M. Cancer stem cells-emanated therapy resistance: implications for liposomal drug delivery systems. J Control Release. 2018;288:62–83 https://doi.org/10.1016/j.jconrel.2018.08.043.PubMedCrossRef

13.

Wang SS, Gao XL, Liu X, Gao SY, Fan YL, Jiang YP, Ma XR, Jiang J, Feng H, Chen QM, Tang YJ, Tang YL, Liang XH. CD133+ cancer stem-like cells promote migration and invasion of salivary adenoid cystic carcinoma by inducing vasculogenic mimicry formation. Oncotarget. 2016;7(20):29051–62 https://doi.org/10.18632/oncotarget.8665.PubMedPubMedCentralCrossRef

14.

Fathi F, Rezabakhsh A, Rahbarghazi R, Rashidi MR. Early-stage detection of VE-cadherin during endothelial differentiation of human mesenchymal stem cells using SPR biosensor. Biosens Bioelectron. 2017;96:358–66 https://doi.org/10.1016/j.bios.2017.05.018.PubMedCrossRef

15.

Lee CH, Wu YT, Hsieh HC, Yu Y, Yu AL, Chang WW. Epidermal growth factor/heat shock protein 27 pathway regulates vasculogenic mimicry activity of breast cancer stem/progenitor cells. Biochimie. 2014;104:117–26 https://doi.org/10.1016/j.biochi.2014.06.011.PubMedCrossRef

16.

Qiao L, Liang N, Zhang J, Xie J, Liu F, Xu D, Yu X, Tian Y. Advanced research on vasculogenic mimicry in cancer. J Cell Mol Med. 2015;19(2):315–26 https://doi.org/10.1111/jcmm.12496.PubMedPubMedCentralCrossRef

17.

Seftor RE, Hess AR, Seftor EA, Kirschmann DA, Hardy KM, Margaryan NV, Hendrix MJ. Tumor cell vasculogenic mimicry: from controversy to therapeutic promise. Am J Pathol. 2012;181(4):1115–25 https://doi.org/10.1016/j.ajpath.2012.07.013.PubMedPubMedCentralCrossRef

18.

Pouyafar A, Heydarabad MZ, Abdolalizadeh J, Zade JA, Rahbarghazi R, Talebi M. Modulation of lipolysis and glycolysis pathways in cancer stem cells changed multipotentiality and differentiation capacity toward endothelial lineage. Cell Biosci. 2019;9:30 https://doi.org/10.1186/s13578-019-0293-z.PubMedPubMedCentralCrossRef

19.

Peng L, Liu A, Shen Y, Xu HZ, Yang SZ, Ying XZ, Liao W, Liu HX, Lin ZQ, Chen QY, Cheng SW, Shen WD. Antitumor and anti-angiogenesis effects of thymoquinone on osteosarcoma through the NF-kappaB pathway. Oncol Rep. 2013;29(2):571–8 https://doi.org/10.3892/or.2012.2165.PubMedCrossRef

20.

ElKhoely A, Hafez HF, Ashmawy AM, Badary O, Abdelaziz A, Mostafa A, Shouman SA. Chemopreventive and therapeutic potentials of thymoquinone in HepG2 cells: mechanistic perspectives. J Nat Med. 2015;69(3):313–23 https://doi.org/10.1007/s11418-015-0895-7.PubMedCrossRef

21.

Paramasivam A, Kalaimangai M, Sambantham S, Anandan B, Jayaraman G. Anti-angiogenic activity of thymoquinone by the down-regulation of VEGF using zebrafish (Danio rerio) model. Biomed Prev Nutr. 2012;2(3):169–73 https://doi.org/10.1016/j.bionut.2012.03.011.CrossRef

22.

Abdelwahab SI, Sheikh BY, Taha MM, How CW, Abdullah R, Yagoub U, El-Sunousi R, Eid EE. Thymoquinone-loaded nanostructured lipid carriers: preparation, gastroprotection, in vitro toxicity, and pharmacokinetic properties after extravascular administration. Int J Nanomedicine. 2013;8:2163–72 https://doi.org/10.2147/IJN.S44108.PubMedPubMedCentralCrossRef

23.

Elmowafy M, Samy A, Raslan MA, Salama A, Said RA, Abdelaziz AE, El-Eraky W, El Awdan S, Viitala T. Enhancement of bioavailability and Pharmacodynamic effects of Thymoquinone via nanostructured lipid carrier (NLC) formulation. AAPS PharmSciTech. 2016;17(3):663–72 https://doi.org/10.1208/s12249-015-0391-0.PubMedCrossRef

24.

Fraveto A, Cardinale V, Bragazzi MC, Giuliante F, De Rose AM, Grazi GL, Napoletano C, Semeraro R, Lustri AM, Costantini D, Nevi L, Di Matteo S, Renzi A, Carpino G, Gaudio E, Alvaro D. Sensitivity of human intrahepatic Cholangiocarcinoma subtypes to chemotherapeutics and molecular targeted agents: a study on primary cell cultures. PLoS One. 2015;10(11):e0142124 https://doi.org/10.1371/journal.pone.0142124.PubMedPubMedCentralCrossRef

25.

Iskender B, Izgi K, Canatan H. Novel anti-cancer agent myrtucommulone-a and thymoquinone abrogate epithelial-mesenchymal transition in cancer cells mainly through the inhibition of PI3K/AKT signalling axis. Mol Cell Biochem. 2016;416(1–2):71–84 https://doi.org/10.1007/s11010-016-2697-y.PubMedCrossRef

26.

Rezabakhsh A, Montazersaheb S, Nabat E, Hassanpour M, Montaseri A, Malekinejad H, Movassaghpour AA, Rahbarghazi R, Garjani A. Effect of hydroxychloroquine on oxidative/nitrosative status and angiogenesis in endothelial cells under high glucose condition. Bioimpacts. 2017;7(4):219–26 https://doi.org/10.15171/bi.2017.26.PubMedPubMedCentralCrossRef

27.

Kundu J, Choi BY, Jeong CH, Kundu JK, Chun KS. Thymoquinone induces apoptosis in human colon cancer HCT116 cells through inactivation of STAT3 by blocking JAK2- and Srcmediated phosphorylation of EGF receptor tyrosine kinase. Oncol Rep. 2014;32(2):821–8 https://doi.org/10.3892/or.2014.3223.PubMedCrossRef

28.

Sethi G, Ahn KS, Aggarwal BB. Targeting nuclear factor-kappa B activation pathway by thymoquinone: role in suppression of antiapoptotic gene products and enhancement of apoptosis. Mol Cancer Res. 2008;6(6):1059–70 https://doi.org/10.1158/1541-7786.MCR-07-2088.PubMedCrossRef

29.

Yi T, Cho SG, Yi Z, Pang X, Rodriguez M, Wang Y, Sethi G, Aggarwal BB, Liu M. Thymoquinone inhibits tumor angiogenesis and tumor growth through suppressing AKT and extracellular signal-regulated kinase signaling pathways. Mol Cancer Ther. 2008;7(7):1789–96 https://doi.org/10.1158/1535-7163.MCT-08-0124.PubMedPubMedCentralCrossRef

30.

Begicevic RR. Falasca M. ABC transporters in Cancer stem cells: beyond Chemoresistance. Int J Mol Sci. 2017;18(11) https://doi.org/10.3390/ijms18112362.

31.

Polakis P. Wnt signaling in cancer. Cold Spring Harb Perspect Biol. 2012;4(5) https://doi.org/10.1101/cshperspect.a008052.

32.

Shi X, Wang J, Lei Y, Cong C, Tan D, Zhou X. Research progress on the PI3K/AKT signaling pathway in gynecological cancer (review). Mol Med Rep. 2019;19(6):4529–35 https://doi.org/10.3892/mmr.2019.10121.PubMedPubMedCentral

33.

Banerjee S, Padhye S, Azmi A, Wang Z, Philip PA, Kucuk O, Sarkar FH, Mohammad RM. Review on molecular and therapeutic potential of thymoquinone in cancer. Nutr Cancer. 2010;62(7):938–46 https://doi.org/10.1080/01635581.2010.509832.PubMedPubMedCentralCrossRef

34.

Moitra K. Overcoming multidrug resistance in Cancer stem cells. Biomed Res Int. 2015;2015:635745 https://doi.org/10.1155/2015/635745.PubMedPubMedCentralCrossRef

35.

Park JW, Jung KH, Byun Y, Lee JH, Moon SH, Cho YS, Lee KH. ATP-binding cassette transporters substantially reduce estimates of ALDH-positive Cancer cells based on Aldefluor and AldeRed588 assays. Sci Rep. 2019;9(1):6462 https://doi.org/10.1038/s41598-019-42954-9.PubMedPubMedCentralCrossRef

36.

Ashour AE, Abd-Allah AR, Korashy HM, Attia SM, Alzahrani AZ, Saquib Q, Bakheet SA, Abdel-Hamied HE, Jamal S, Rishi AK. Thymoquinone suppression of the human hepatocellular carcinoma cell growth involves inhibition of IL-8 expression, elevated levels of TRAIL receptors, oxidative stress and apoptosis. Mol Cell Biochem. 2014;389(1–2):85–98 https://doi.org/10.1007/s11010-013-1930-1.PubMedCrossRef

37.

Mostofa AGM, Hossain MK, Basak D, Bin Sayeed MS. Thymoquinone as a potential adjuvant therapy for Cancer treatment: evidence from preclinical studies. Front Pharmacol. 2017;8:295 https://doi.org/10.3389/fphar.2017.00295.PubMedPubMedCentralCrossRef

38.

Dean M, Fojo T, Bates S. Tumour stem cells and drug resistance. Nat Rev Cancer. 2005;5(4):275–84 https://doi.org/10.1038/nrc1590.PubMedCrossRef

39.

Jiang ZS, Sun YZ, Wang SM, Ruan JS. Epithelial-mesenchymal transition: potential regulator of ABC transporters in tumor progression. J Cancer. 2017;8(12):2319–27 https://doi.org/10.7150/jca.19079.PubMedPubMedCentralCrossRef

40.

Lang M, Borgmann M, Oberhuber G, Evstatiev R, Jimenez K, Dammann KW, Jambrich M, Khare V, Campregher C, Ristl R, Gasche C. Thymoquinone attenuates tumor growth in ApcMin mice by interference with Wnt-signaling. Mol Cancer. 2013;12(1):41 https://doi.org/10.1186/1476-4598-12-41.PubMedPubMedCentralCrossRef

41.

Ndreshkjana B, Capci A, Klein V, Chanvorachote P, Muenzner JK, Huebner K, Steinmann S, Erlenbach-Wuensch K, Geppert CI, Agaimy A, Ballout F, El-Baba C, Gali-Muhtasib H, Roehe AV, Hartmann A, Tsogoeva SB, Schneider-Stock R. Combination of 5-fluorouracil and thymoquinone targets stem cell gene signature in colorectal cancer cells. Cell Death Dis. 2019;10(6):379 https://doi.org/10.1038/s41419-019-1611-4.PubMedPubMedCentralCrossRef

42.

Rajput S, Kumar BN, Sarkar S, Das S, Azab B, Santhekadur PK, Das SK, Emdad L, Sarkar D, Fisher PB, Mandal M. Targeted apoptotic effects of thymoquinone and tamoxifen on XIAP mediated Akt regulation in breast cancer. PLoS One. 2013;8(4):e61342 https://doi.org/10.1371/journal.pone.0061342.PubMedPubMedCentralCrossRef

43.

Xu D, Ma Y, Zhao B, Li S, Zhang Y, Pan S, Wu Y, Wang J, Wang D, Pan H, Liu L, Jiang H. Thymoquinone induces G2/M arrest, inactivates PI3K/Akt and nuclear factor-kappaB pathways in human cholangiocarcinomas both in vitro and in vivo. Oncol Rep. 2014;31(5):2063–70 https://doi.org/10.3892/or.2014.3059.PubMedCrossRef

44.

Mojzis J, Varinska L, Mojzisova G, Kostova I, Mirossay L. Antiangiogenic effects of flavonoids and chalcones. Pharmacol Res. 2008;57(4):259–65 https://doi.org/10.1016/j.phrs.2008.02.005.PubMedCrossRef

45.

Pouyafar A, Rezabakhsh A, Rahbarghazi R, Heydarabad MZ, Shokrollahi E, Sokullu E, Khaksar M, Nourazarian A, Avci ÇB. Treatment of cancer stem cells from human colon adenocarcinoma cell line HT-29 with resveratrol and sulindac induced mesenchymal-endothelial transition rate. Cell Tissue Res. 2019;376(3):377–88 https://doi.org/10.1007/s00441-019-02998-9.PubMedCrossRef

46.

Kim HS, Won YJ, Shim JH, Kim HJ, Kim BS, Hong HN. Role of EphA2-PI3K signaling in vasculogenic mimicry induced by cancer-associated fibroblasts in gastric cancer cells. Oncol Lett. 2019;18(3):3031–8 https://doi.org/10.3892/ol.2019.10677.PubMedPubMedCentral

47.

Lu XS, Sun W, Ge CY, Zhang WZ, Fan YZ. Contribution of the PI3K/MMPs/Ln-5γ2 and EphA2/FAK/Paxillin signaling pathways to tumor growth and vasculogenic mimicry of gallbladder carcinomas. Int J Oncol. 2013;42(6):2103–15 https://doi.org/10.3892/ijo.2013.1897.PubMedCrossRef

Title: Thymoquinone inhibited vasculogenic capacity and promoted mesenchymal-epithelial transition of human breast cancer stem cells
Authors: Sanya Haiaty
Mohammad-Reza Rashidi
Maryam Akbarzadeh
Ahad Bazmani
Mostafa Mostafazadeh
Saba Nikanfar
Zohre Zibaei
Reza Rahbarghazi
Mohammad Nouri
Publication date: 01-12-2021
Publisher: BioMed Central
Keywords: Breast Cancer
Breast Cancer
Published in: BMC Complementary Medicine and Therapies / Issue 1/2021
Electronic ISSN: 2662-7671
DOI: https://doi.org/10.1186/s12906-021-03246-w

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Thymoquinone inhibited vasculogenic capacity and promoted mesenchymal-epithelial transition of human breast cancer stem cells

Abstract

Background

Methods

Results

Conclusion

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 1/2021

Piperlongumine inhibits the growth of non-small cell lung cancer cells via the miR-34b-3p/TGFBR1 pathway

A systems pharmacology approach based on oncogenic signalling pathways to determine the mechanisms of action of natural products in breast cancer from transcriptome data

Problems with the outcome measures in randomized controlled trials of traditional Chinese medicine in treating chronic heart failure caused by coronary heart disease: a systematic review

Protective effect of 6-paradol in acetic acid-induced ulcerative colitis in rats

The use of complementary and alternative medicine during pregnancy: a cross-sectional study from Palestine

The use of complementary and alternative medicine by adults with allergies: a Czech national representative survey