nach oben

Journal of Natural Medicines

Erschienen in:

01.10.2015 | Original Paper

Cucurbitacin B induces DNA damage and autophagy mediated by reactive oxygen species (ROS) in MCF-7 breast cancer cells

verfasst von: Guowen Ren, Tongye Sha, Jiajie Guo, Wenxue Li, Jinjian Lu, Xiuping Chen

Erschienen in: Journal of Natural Medicines | Ausgabe 4/2015

Einloggen, um Zugang zu erhalten

Abstract

Cucurbitacin B (Cuc B), a natural compound extracted from cucurbitaceous plants, demonstrated potent anticancer activities, while the underlying mechanisms remain unclear. We investigated the anticancer effect of Cuc B on MCF-7 breast cancer cells. Cuc B drastically decreased cell viability in a concentration-dependent manner. Cuc B treatment caused DNA damage, as shown by long tails in the comet assay and increased γH₂AX protein expression. Immunofluorescence staining showed that Cuc B treatment induced nuclear γH₂AX foci. Cuc B activated DNA damage pathways by phosphorylation of ATM/ATR [two large phosphatidylinositol-3-kinase-like kinase family (PIKKs) members]. Furthermore, it also induced autophagy, as evidenced by monodansylcadaverine (MDC) staining and autophagic protein expression. In addition, Cuc B treatment led to increased reactive oxygen species (ROS) formation, which was inhibited by N-acetyl-l-cysteine (NAC) pretreatment. NAC pretreatment inhibited Cuc-B-induced DNA damage and autophagy. Taken together, these results suggest that ROS-mediated Cuc-B-induced DNA damage and autophagy in MCF-7 cells, which provides new insights into the anticancer molecular mechanism of Cuc B.

Chen JC, Chiu MH, Nie RL, Cordell GA, Qiu SX (2005) Cucurbitacins and cucurbitane glycosides: structures and biological activities. Nat Prod Rep 22:386–399CrossRefPubMed

Chen X, Bao J, Guo J, Ding Q, Lu J, Huang M, Wang Y (2012) Biological activities and potential molecular targets of cucurbitacins: a focus on cancer. Anticancer Drugs 23:777–787CrossRefPubMed

Liu T, Zhang M, Zhang H, Sun C, Deng Y (2008) Inhibitory effects of cucurbitacin B on laryngeal squamous cell carcinoma. Eur Arch Otorhinolaryngol 265:1225–1232CrossRefPubMed

Thoennissen NH, Iwanski GB, Doan NB, Okamoto R, Lin P, Abbassi S, Song JH, Yin D, Toh M, Xie WD (2009) Cucurbitacin B induces apoptosis by inhibition of the JAK/STAT pathway and potentiates antiproliferative effects of gemcitabine on pancreatic cancer cells. Cancer Res 69:5876–5884CrossRefPubMed

Zhang M, Zhang H, Sun C, Shan X, Yang X, Li-Ling J, Deng Y (2009) Targeted constitutive activation of signal transducer and activator of transcription 3 in human hepatocellular carcinoma cells by cucurbitacin B. Cancer Chemother Pharmacol 63:635–642CrossRefPubMed

Chan KT, Meng FY, Li Q, Ho CY, Lam TS, To Y, Lee WH, Li M, Chu KH, Toh M (2010) Cucurbitacin B induces apoptosis and S phase cell cycle arrest in BEL-7402 human hepatocellular carcinoma cells and is effective via oral administration. Cancer Lett 294:118–124CrossRefPubMed

Zhang Y, Ouyang D, Xu L, Ji Y, Zha Q, Cai J, He X (2011) Cucurbitacin B induces rapid depletion of the G-actin pool through reactive oxygen species-dependent actin aggregation in melanoma cells. Acta Biochim Biophys Sin (Shanghai) 43:556–567CrossRef

Kausar H, Munagala R, Bansal SS, Aqil F, Vadhanam MV, Gupta RC (2013) Cucurbitacin B potently suppresses non-small-cell lung cancer growth: identification of intracellular thiols as critical targets. Cancer Lett 332:35–45CrossRefPubMed

Liu T, Peng H, Zhang M, Deng Y, Wu Z (2010) Cucurbitacin B, a small molecule inhibitor of the Stat3 signaling pathway, enhances the chemosensitivity of laryngeal squamous cell carcinoma cells to cisplatin. Eur J Pharmacol 641:15–22CrossRefPubMed

10.

Iwanski GB, Lee DH, En-Gal S, Doan NB, Castor B, Vogt M, Toh M, Bokemeyer C, Said JW, Thoennissen NH (2010) Cucurbitacin B, a novel in vivo potentiator of gemcitabine with low toxicity in the treatment of pancreatic cancer. Br J Pharmacol 160:998–1007PubMedCentralCrossRefPubMed

11.

Lee DH, Thoennissen NH, Goff C, Iwanski GB, Forscher C, Doan NB, Said JW, Phillip Koeffler H (2011) Synergistic effect of low-dose cucurbitacin B and low-dose methotrexate for treatment of human osteosarcoma. Cancer Lett 306:161–170CrossRefPubMed

12.

Aribi A, Gery S, Lee DH, Thoennissen NH, Thoennissen GB, Alvarez R, Ho Q, Lee K, Doan NB, Chan KT (2013) The triterpenoid cucurbitacin B augments the antiproliferative activity of chemotherapy in human breast cancer. Int J Cancer 132:2730–2737PubMedCentralCrossRefPubMed

13.

Wakimoto N, Yin D, O’Kelly J, Haritunians T, Karlan B, Said J, Xing H, Koeffler HP (2008) Cucurbitacin B has a potent antiproliferative effect on breast cancer cells in vitro and in vivo. Cancer Sci 99:1793–1797CrossRefPubMed

14.

Gupta P, Srivastava SK (2014) Inhibition of Integrin-HER2 signaling by Cucurbitacin B leads to in vitro and in vivo breast tumor growth suppression. Oncotarget 5:1812–1828PubMedCentralPubMed

15.

Duangmano S, Sae-Lim P, Suksamrarn A, Domann FE, Patmasiriwat P (2012) Cucurbitacin B inhibits human breast cancer cell proliferation through disruption of microtubule polymerization and nucleophosmin/B23 translocation. BMC Complement Altern Med 12:185PubMedCentralCrossRefPubMed

16.

Duangmano S, Dakeng S, Jiratchariyakul W, Suksamrarn A, Smith DR, Patmasiriwat P (2010) Antiproliferative effects of cucurbitacin B in breast cancer cells: down-regulation of the c-Myc/hTERT/telomerase pathway and obstruction of the cell cycle. Int J Mol Sci 11:5323–5338PubMedCentralCrossRefPubMed

17.

18.

Dakeng S, Duangmano S, Jiratchariyakul W, Up Y, Bogler O, Patmasiriwat P (2012) Inhibition of Wnt signaling by cucurbitacin B in breast cancer cells: reduction of Wnt-associated proteins and reduced translocation of galectin-3-mediated beta-catenin to the nucleus. J Cell Biochem 113:49–60PubMedCentralCrossRefPubMed

19.

Ray PD, Huang B-W, Tsuji Y (2012) Reactive oxygen species (ROS) homeostasis and redox regulation in cellular signaling. Cell Signal 24:981–990PubMedCentralCrossRefPubMed

20.

Zhang T, Li Y, Park KA, Byun HS, Won M, Jeon J, Lee Y, Seok JH, Choi S-W, Lee S-H (2012) Cucurbitacin induces autophagy through mitochondrial ROS production which counteracts to limit caspase-dependent apoptosis. Autophagy 8:559–576CrossRefPubMed

21.

Yasuda S, Yogosawa S, Izutani Y, Nakamura Y, Watanabe H, Sakai T (2010) Cucurbitacin B induces G2 arrest and apoptosis via a reactive oxygen species-dependent mechanism in human colon adenocarcinoma SW480 cells. Mol Nutr Food Res 54:559–565CrossRefPubMed

22.

Filomeni G, De Zio D, Cecconi F (2014) Oxidative stress and autophagy: the clash between damage and metabolic needs. Cell Death Differ 22:377–388PubMedCentralCrossRefPubMed

23.

Guo J, Wu G, Bao J, Hao W, Lu J, Chen X (2014) Cucurbitacin B induced ATM-mediated DNA damage causes G2/M cell cycle arrest in a ROS-dependent manner. PLoS One 9:e88140PubMedCentralCrossRefPubMed

24.

Guo J, Zhao W, Hao W, Ren G, Lu J, Chen X (2014) Cucurbitacin B induces DNA damage, G2/M phase arrest, and apoptosis mediated by reactive oxygen species (ROS) in leukemia K562 cells. Anticancer Agents Med Chem 14:1146–1153CrossRefPubMed

25.

Munafo DB, Colombo MI (2001) A novel assay to study autophagy: regulation of autophagosome vacuole size by amino acid deprivation. J Cell Sci 114:3619–3629PubMed

26.

Lord CJ, Ashworth A (2012) The DNA damage response and cancer therapy. Nature 481:287–294CrossRefPubMed

27.

Garcia-Canton C, Anadon A, Meredith C (2012) gammaH2AX as a novel endpoint to detect DNA damage: applications for the assessment of the in vitro genotoxicity of cigarette smoke. Toxicol In Vitro 26:1075–1086CrossRefPubMed

28.

Mah LJ, El-Osta A, Karagiannis TC (2010) gammaH2AX: a sensitive molecular marker of DNA damage and repair. Leukemia 24:679–686CrossRefPubMed

29.

Maréchal A, Zou L (2013) DNA damage sensing by the ATM and ATR kinases. Cold Spring Harb Perspect Biol 5(9)

30.

Smith J, Tho LM, Xu N, Gillespie DA (2010) The ATM-Chk2 and ATR-Chk1 pathways in DNA damage signaling and cancer. Adv Cancer Res 108:73–112CrossRefPubMed

31.

Nagelkerke A, Bussink J, Geurts-Moespot A, Sweep FC, Span PN (2014) Therapeutic targeting of autophagy in cancer. Part II: pharmacological modulation of treatment-induced autophagy. Semin Cancer Biol 31:99–105CrossRefPubMed

32.

Thorburn A, Thamm DH, Gustafson DL (2014) Autophagy and cancer therapy. Mol Pharmacol 85:830–838PubMedCentralCrossRefPubMed

33.

Zhang T, Li Y, Park KA, Byun HS, Won M, Jeon J, Lee Y, Seok JH, Choi SW, Lee SH, Man Kim J, Lee JH, Son CG, Lee ZW, Shen HM, Hur GM (2012) Cucurbitacin induces autophagy through mitochondrial ROS production which counteracts to limit caspase-dependent apoptosis. Autophagy 8:559–576CrossRefPubMed

34.

He J, Wang Y, Xu LH, Qiao J, Ouyang DY, He XH (2013) Cucurbitacin IIa induces caspase-3-dependent apoptosis and enhances autophagy in lipopolysaccharide-stimulated RAW 264.7 macrophages. Int Immunopharmacol 16:27–34CrossRefPubMed

35.

Zhu JS, Ouyang DY, Shi ZJ, Xu LH, Zhang YT, He XH (2012) Cucurbitacin B induces cell cycle arrest, apoptosis and autophagy associated with G actin reduction and persistent activation of cofilin in Jurkat cells. Pharmacology 89:348–356CrossRefPubMed

36.

Yuan G, Yan SF, Xue H, Zhang P, Sun JT, Li G (2014) Cucurbitacin I induces protective autophagy in glioblastoma in vitro and in vivo. J Biol Chem 289:10607–10619PubMedCentralCrossRefPubMed

37.

Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, Kominami E, Ohsumi Y, Yoshimori T (2000) LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J 19:5720–5728PubMedCentralCrossRefPubMed

38.

Mizushima N, Yoshimori T, Levine B (2010) Methods in mammalian autophagy research. Cell 140:313–326PubMedCentralCrossRefPubMed

39.

Vázquez CL, Colombo MI. (2009) Chapter 6 assays to assess autophagy induction and fusion of autophagic vacuoles with a degradative compartment, using monodansylcadaverine (MDC) and DQ‐BSA. In: Daniel JK (ed) Methods in enzymology. Academic Press, pp 85–95

40.

Rubinsztein DC, Codogno P, Levine B (2012) Autophagy modulation as a potential therapeutic target for diverse diseases. Nat Rev Drug Discov 11:709–730PubMedCentralCrossRefPubMed

41.

Kang R, Zeh HJ, Lotze MT, Tang D (2011) The Beclin 1 network regulates autophagy and apoptosis. Cell Death Differ 18:571–580PubMedCentralCrossRefPubMed

42.

Myeku N, Figueiredo-Pereira ME (2011) Dynamics of the degradation of ubiquitinated proteins by proteasomes and autophagy: association with sequestosome 1/p62. J Biol Chem 286:22426–22440PubMedCentralCrossRefPubMed

43.

Scherz-Shouval R, Elazar Z (2007) ROS, mitochondria and the regulation of autophagy. Trends Cell Biol 17:422–427CrossRefPubMed

44.

Azad MB, Chen Y, Gibson SB (2009) Regulation of autophagy by reactive oxygen species (ROS): implications for cancer progression and treatment. Antioxid Redox Signal 11:777–790CrossRefPubMed

45.

Kurz EU, Lees-Miller SP (2004) DNA damage-induced activation of ATM and ATM-dependent signaling pathways. DNA Repair 3:889–900CrossRefPubMed

46.

Titel: Cucurbitacin B induces DNA damage and autophagy mediated by reactive oxygen species (ROS) in MCF-7 breast cancer cells
verfasst von: Guowen Ren
Tongye Sha
Jiajie Guo
Wenxue Li
Jinjian Lu
Xiuping Chen
Publikationsdatum: 01.10.2015
Verlag: Springer Japan
Erschienen in: Journal of Natural Medicines / Ausgabe 4/2015
Print ISSN: 1340-3443
Elektronische ISSN: 1861-0293
DOI: https://doi.org/10.1007/s11418-015-0918-4

Springer Medizin

Abstract

Bitte loggen Sie sich ein, um Zugang zu diesem Inhalt zu erhalten

Weitere Artikel der Ausgabe 4/2015

Dipeptidyl peptidase-IV inhibitory activity of dimeric dihydrochalcone glycosides from flowers of Helichrysum arenarium

Myrtucommulone-A treatment decreases pluripotency- and multipotency-associated marker expression in bladder cancer cell line HTB-9

New norditerpenoid alkaloids from Aconitum vilmorinianum Komarov

Acyl flavonoids, biflavones, and flavonoids from Cephalotaxus harringtonia var. nana

Hedgehog/GLI-mediated transcriptional activity inhibitors from Crinum asiaticum

Chikungunya virus (CHIKV) inhibitors from natural sources: a medicinal chemistry perspective