Advertisement

Celastrol enhances TRAIL-induced apoptosis in human glioblastoma via the death receptor pathway

  • Zhe Cha
  • Jianzhang Cheng
  • Hui Xiang
  • Jingjing Qin
  • Yujia He
  • Zhiping Peng
  • Jianhua Jia
  • Huarong YuEmail author
Original Article
  • 34 Downloads

Abstract

Purpose

Glioblastoma is the most common, malignant and devastating type of primary brain tumor. Tumor necrosis factor-related apoptosis-induced ligand (TRAIL) is characterized by its lethality to precancerous and cancerous cells. However, many kinds of tumor cells, including most glioma cells, tend to evade TRAIL-induced apoptosis. Celastrol is a pleiotropic compound from a traditional Chinese medicine that has proven to be useful as a sensitizer for TRAIL treatment. However, the underlying mechanism and role of celastrol in the sensitization of glioma cells remain to be elucidated.

Methods

The viability of glioma cell lines was examined by the CCK-8 assay. The expression of DR5 was detected by reverse transcriptase quantitative real-time PCR. The protein expression of DR5, cleaved caspase-8, cleaved caspase-3 and PARP were measured by western blot. The apoptosis rates and the sub-G1 population were detected by flow cytometry. The cellular morphological changes were assessed by TUNEL apoptosis and Hoechst 33258 staining assays. The knockdown of DR5 expression was conducted by siRNA.

Results

In this study, we observed that celastrol treatment inhibited cell viability in a dose-dependent manner, while glioma and normal human astroglial cell lines were resistant to TRAIL treatment. We also observed that the antiproliferative effects of TRAIL in combination with a noncytotoxic concentration of celastrol were significantly greater than those of celastrol or TRAIL alone. In addition, cell death induced by the combination treatment was apoptotic and occurred through the death receptor pathway via activation of caspase-8, caspase-3, and PARP. Furthermore, celastrol upregulated death receptor 5 (DR5) at the mRNA and protein levels, and siRNA-mediated DR5 knockdown reduced the killing effect of the combination drug treatment on glioma cells and reduced the activation of caspase-3, caspase-8 and PARP.

Conclusions

Taken together, the results of our study demonstrate that celastrol sensitizes glioma cells to TRAIL via the death receptor pathway and that DR5 plays an important role in the effects of this cotreatment. The results indicate that this cotreatment is a promising tumor-killing therapeutic strategy with high efficacy and low toxicity.

Keywords

Celastrol TRAIL U87-MG DR5 Apoptosis 

Abbreviations

TRAIL

Tumor necrosis factor-related apoptosis-induced ligand

DR5

Death receptor 5

FBS

Fetal bovine serum

PARP

Poly-ADP-ribose polymerase

CNS

Central nervous system

GBM

Glioblastoma multiforme

DR4

Death receptor 4

FADD

Fas-associated death domain

DISC

Death-inducing signaling complex

DMSO

Dimethylsulfoxide

STR

Short tandem repeat

CCK-8

Cell Counting Kit-8

FCM

Flow cytometry

SiRNA

Small interfering RNA

PVDF

Polyvinylidene fluoride

TBST

Tris-buffered saline with Tween

qRT-PCR

Reverse transcriptase quantitative real-time PCR

DcR

Decoy receptors

CDI

Coefficient of drug interaction

Notes

Funding

This study was funded by the Chongqing Fundamental Research Funds for nonprofit public scientific research institutions from the Chongqing Science and Technology Commission (Grant number 2015CSTC-JBKY-01702).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors. The manuscript does not contain clinical studies or patient data.

References

  1. 1.
    Louis DN, Perry A, Reifenberger G, von Deimling A, Figarella-Branger D, Cavenee WK, Ohgaki H, Wiestler OD, Kleihues P, Ellison DW (2016) The 2016 World Health Organization classification of tumors of the central nervous system: a summary. Acta Neuropathol 131:803–820.  https://doi.org/10.1007/s00401-016-1545-1 CrossRefGoogle Scholar
  2. 2.
    Wiley SR, Schooley K, Smolak PJ, Din WS, Huang CP, Nicholl JK, Sutherland GF, Smith TD, Rauch C, Smith CA, Goodwin RG (1995) Identification and characterization of a new member of the TNF family that induces apoptosis. Immunity 3:673–682.  https://doi.org/10.1016/1074-7613(95)90057-8 CrossRefGoogle Scholar
  3. 3.
    Sheridan JP, Marsters SA, Pitti RM, Gurney A, Skubatch M, Baldwin D, Ramakrishnan L, Gray CL, Baker K, Wood WI, Goddard AD, Godowski P, Ashkenazi A (1997) Control of TRAIL-induced apoptosis by a family of signaling and decoy receptors. Science 277:818–821.  https://doi.org/10.1126/science.277.5327.818 CrossRefGoogle Scholar
  4. 4.
    Pan G, Ni J, Wei Y-F, Yu G, Gentz R, Dixit VM (1997) An antagonist decoy receptor and a death domain-containing receptor for TRAIL. Science 277:815–818.  https://doi.org/10.1126/science.277.5327.815 CrossRefGoogle Scholar
  5. 5.
    Wang S, El-Deiry WS (2003) TRAIL and apoptosis induction by TNF-family death receptors. Oncogene 22:8628–8633.  https://doi.org/10.1038/sj.onc.1207232 CrossRefGoogle Scholar
  6. 6.
    Hao C, Beguinot F, Condorelli G, Trencia A, Van Meir EG, Yong VW, Parney IF, Roa WH, Petruk KCJCR (2001) Induction and intracellular regulation of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) mediated apoptosis in human malignant glioma cells. Cancer Res 61:1162–1170.  https://doi.org/10.1097/00002820-200102000-00011 Google Scholar
  7. 7.
    Siegelin MD, Reuss DE, Habel A, Rami A, von Deimling A (2009) Quercetin promotes degradation of survivin and thereby enhances death-receptor-mediated apoptosis in glioma cells. Neuro Oncol 11:122–131.  https://doi.org/10.1215/152285172008-085 CrossRefGoogle Scholar
  8. 8.
    Zhu H, Liu XW, Ding WJ, Xu DQ, Zhao YC, Lu W, He QJ, Yang B (2010) Up-regulation of death receptor 4 and 5 by celastrol enhances the anti-cancer activity of TRAIL/Apo-2L. Cancer Lett 297:155–164.  https://doi.org/10.1016/j.canlet.2010.04.030 CrossRefGoogle Scholar
  9. 9.
    Jin CY, Park C, Hwang HJ, Kim GY, Choi BT, Kim WJ, Choi YH (2011) Naringenin up-regulates the expression of death receptor 5 and enhances TRAIL-induced apoptosis in human lung cancer A549 cells. Mol Nutr Food Res 55:300–309.  https://doi.org/10.1002/mnfr.201000024 CrossRefGoogle Scholar
  10. 10.
    Hwang JS, Lee YY, Lee DH, Kwon KH (2017) DATS sensitizes glioma cells to TRAIL-mediated apoptosis by up-regulation of death receptor 5 via ROS. Food Chem Toxicol 106:514–521.  https://doi.org/10.1016/j.fct.2017.05.056 CrossRefGoogle Scholar
  11. 11.
    Ashkenaz A, Dixit VM (1998) Death receptors: signaling and modulation. Science 281:1305–1308.  https://doi.org/10.1126/science.281.5381.1305 CrossRefGoogle Scholar
  12. 12.
    Wajant H, Gerspach J, Pfizenmaier K (2005) Tumor therapeutics by design: targeting and activation of death receptors. Cytokine Growth Factor Rev 16:55–76.  https://doi.org/10.1016/j.cytogfr.2004.12.001 CrossRefGoogle Scholar
  13. 13.
    Pinna GF, Fiorucci M, Reimund JM, Taquet N, Arondel Y, Muller CD (2004) Celastrol inhibits pro-inflammatory cytokine secretion in Crohn’s disease biopsies. Biochem Biophys Res Commun 322:778–786.  https://doi.org/10.1016/j.bbrc.2004.07.186 CrossRefGoogle Scholar
  14. 14.
    Zhao J, Sun Y, Shi P, Dong JN, Zuo LG, Wang HG, Gong JF, Li Y, Gu LL, Li N, Li JS, Zhu WM (2015) Celastrol ameliorates experimental colitis in IL-10 deficient mice via the up-regulation of autophagy. Int Immunopharmacol 26:221–228.  https://doi.org/10.1016/j.intimp.2015.03.033 CrossRefGoogle Scholar
  15. 15.
    Xin W, Wang Q, Zhang D, Wang C (2017) A new mechanism of inhibition of IL-1beta secretion by celastrol through the NLRP3 inflammasome pathway. Eur J Pharmacol 814:240–247.  https://doi.org/10.1016/j.ejphar.2017.08.036 CrossRefGoogle Scholar
  16. 16.
    Allison AC, Cacabelos R, Lombardi VR, Alvarez XA, Vigo C (2001) Celastrol, a potent antioxidant and anti-inflammatory drug, as a possible treatment for Alzheimer’s disease. Prog Neuro Psychopharmacol Biol Psychiatry 25:1341–1357.  https://doi.org/10.1016/S0278-5846(01)00192-0 CrossRefGoogle Scholar
  17. 17.
    Yang H, Chen D, Cui QC, Yuan X, Dou QP (2006) Celastrol, a triterpene extracted from the Chinese “Thunder of God Vine,” is a potent proteasome inhibitor and suppresses human prostate cancer growth in nude mice. Cancer Res 66:4758–4765.  https://doi.org/10.1158/0008-5472.CAN-05-4529 CrossRefGoogle Scholar
  18. 18.
    Lee JH, Won YS, Park KH, Lee MK, Tachibana H, Yamada K, Seo KI (2012) Celastrol inhibits growth and induces apoptotic cell death in melanoma cells via the activation ROS-dependent mitochondrial pathway and the suppression of PI3 K/AKT signaling. Apoptosis 17:1275–1286.  https://doi.org/10.1007/s10495-012-0767-5 CrossRefGoogle Scholar
  19. 19.
    Li HY, Zhang J, Sun LL, Li BH, Gao HL, Xie T, Zhang N, Ye ZM (2015) Celastrol induces apoptosis and autophagy via the ROS/JNK signaling pathway in human osteosarcoma cells: an in vitro and in vivo study. Cell Death Dis 6:e1604.  https://doi.org/10.1038/cddis.2014.543 CrossRefGoogle Scholar
  20. 20.
    Boridy S, Le P, Petrecca K, Maysinger D (2015) Celastrol targets proteostasis and acts synergistically with a heat-shock protein 90 inhibitor to kill human glioblastoma cells. Cell Death Dis 5:e1216.  https://doi.org/10.1038/cddis.2014.182 CrossRefGoogle Scholar
  21. 21.
    Zhu H, Ding WJ, Wu R, Weng QJ, Lou JS, Jin RJ, Lu W, Yang B, He QJ (2010) Synergistic anti-cancer activity by the combination of TRAIL/APO-2L and celastrol. Cancer Invest 28:23–32.  https://doi.org/10.3109/07357900903095664 CrossRefGoogle Scholar
  22. 22.
    Khan M, Bi Y, Qazi JI, Fan L, Gao H (2015) Evodiamine sensitizes U87 glioblastoma cells to TRAIL via the death receptor pathway. Mol Med Rep 11:257–262.  https://doi.org/10.3892/mmr.2014.2705 CrossRefGoogle Scholar
  23. 23.
    Jung EM, Lim JH, Lee TJ, Park JW, Choi KS, Kwon TK (2005) Curcumin sensitizes tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis through reactive oxygen species-mediated upregulation of death receptor 5 (DR5). Carcinogenesis 26:1905–1913.  https://doi.org/10.1093/carcin/bgi167 CrossRefGoogle Scholar
  24. 24.
    Hombach-Klonisch S, Mehrpour M, Shojaei S, Harlos C, Pitz M, Hamai A, Siemianowicz K, Likus W, Wiechec E, Toyota BD, Hoshyar R, Seyfoori A, Sepehri Z, Ande SR, Khadem F, Akbari M, Gorman AM, Samali A, Klonisch T, Ghavami S (2018) Glioblastoma and chemoresistance to alkylating agents: involvement of apoptosis, autophagy, and unfolded protein response. Pharmacol Ther 184:13–41.  https://doi.org/10.1016/j.pharmthera.2017.10.017 CrossRefGoogle Scholar
  25. 25.
    Cuello M, Ettenberg SA, Nau MM, Lipkowitz S (2001) Synergistic induction of apoptosis by the combination of trail and chemotherapy in chemoresistant ovarian cancer cells. Gynecol Oncol 81:380–390.  https://doi.org/10.1006/gyno.2001.6194 CrossRefGoogle Scholar
  26. 26.
    Ray S, Shyam S, Fraizer GC, Almasan A (2007) S-phase checkpoints regulate Apo2 ligand/TRAIL and CPT-11-induced apoptosis of prostate cancer cells. Mol Cancer Ther 6:1368–1378.  https://doi.org/10.1158/1535-7163.MCT-05-0414 CrossRefGoogle Scholar
  27. 27.
    Chen SR, Dai Y, Zhao J, Lin L, Wang Y, Wang Y (2018) A mechanistic overview of triptolide and celastrol, natural products from Tripterygium wilfordii Hook F. Front Pharmacol 9:104.  https://doi.org/10.3389/fphar.2018.00104 CrossRefGoogle Scholar
  28. 28.
    Almasan A, Ashkenazi A (2003) Apo2L/TRAIL: apoptosis signaling, biology, and potential for cancer therapy. Cytokine Growth Factor Rev 14:337–348.  https://doi.org/10.1016/S1359-6101(03)00029-7 CrossRefGoogle Scholar
  29. 29.
    Kuijlen JM, Mooij JJ, Platteel I, Hoving EW, van der Graaf WT, Span MM, Hollema H, den Dunnen WF (2006) TRAIL-receptor expression is an independent prognostic factor for survival in patients with a primary glioblastoma multiforme. J Neuro Oncol 78:161–171.  https://doi.org/10.1007/s11060-005-9081-1 CrossRefGoogle Scholar
  30. 30.
    Mert U, Sanlioglu AD (2017) Intracellular localization of DR5 and related regulatory pathways as a mechanism of resistance to TRAIL in cancer. Mol Life Sci 74:245–255.  https://doi.org/10.1007/s00018-016-2321-z CrossRefGoogle Scholar
  31. 31.
    Hetschko H, Voss V, Horn S, Seifert V, Prehn JH, Kogel D (2008) Pharmacological inhibition of Bcl-2 family members reactivates TRAIL-induced apoptosis in malignant glioma. J Neuro Oncol 86:265–272.  https://doi.org/10.1007/s11060-007-9472-6 CrossRefGoogle Scholar
  32. 32.
    Wang D, Wang Z, Tian B, Li X, Li S, Tian Y (2008) Two hour exposure to sodium butyrate sensitizes bladder cancer to anticancer drugs. Int J Urol 15(5):435–441.  https://doi.org/10.1111/j.1442-2042.2008.02025.x CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Research Center of NeuroscienceChongqing Medical UniversityChongqingChina
  2. 2.Laboratory of Radiological MedicineChongqing Medical UniversityChongqingChina

Personalised recommendations