Resveratrol and siRNA in combination reduces Hsp27 expression and induces caspase-3 activity in human glioblastoma cells

  • Evren Önay UçarEmail author
  • Aslıhan Şengelen
Original Paper


GBM cells can easily gain resistance to conventional therapy, and therefore treatment of glioblastoma multiforme (GBM) is difficult. One of the hallmark proteins known to be responsible for this resistance is heat shock protein 27 (Hsp27) which has a key role in the cell survival. Resveratrol, a natural compound, exhibits antitumor effects against GBM, but there are no reports regarding its effect on Hsp27 expression in gliomas. The aim of the present study was to asses the effect of resveratrol on Hsp27 expression and apoptosis in non-transfected and transfected U-87 MG human glioblastoma cells. In order to block the Hsp27 expression, siRNA transfection was performed. Non-transfected and transfected cells were treated with either 10 or 15 μM resveratrol. The effects of resveratrol were compared with quercetin, a well-known Hsp27 inhibitor. Resveratrol was found to induce apoptosis more effectively than quercetin. Our data showed that resveratrol induces dose- and time-dependent cell death. We also determined that silencing of Hsp27 with siRNA makes the cells more vulnerable to apoptosis upon resveratrol treatment. The highest effect was observed in the 15 μM resveratrol and 25 nM siRNA combination group (suppressed Hsp27 expression by 93.4% and induced apoptosis by 101.2%). This study is the first report showing that resveratrol reduces Hsp27 levels, and siRNA-mediated Hsp27 silencing enhances the therapeutic effects of resveratrol in glioma cells. Our results suggest that resveratrol administration in combination with Hsp27 silencing has a potential to be used as a candidate for GBM treatment.


Resveratrol siRNA Combined therapy Hsp27 Glioma Apoptosis 



Protein kinase B (PKB)


Dimethyl sulfoxide


Endoplasmic reticulum


Glyceraldehyde 3-phosphate dehydrogenase


Glioblastoma multiforme


Heat shock factor


Heat shock protein


The half-maximal inhibitory concentration


Mitogen-activated protein kinase


Mammalian target of rapamycin


3-(4,5-Dimethylthiazol-2-yl) 2, 5-diphenyl-tetrazolium bromide


Nuclear factor kappa B


Phosphatidylinositol 3-kinase


RNA interference


Small interfering RNA

Sirtuin 1



Tumor necrosis factor alpha


TNF-related apoptosis-inducing ligand


Funding information

This work was supported by the Istanbul University Research Foundation, Turkey (Project numbers 57959 and BEK-24987).


  1. Banerjee Mustafi S, Chakraborty PK, Raha S (2010) Modulation of Akt and ERK1/2 pathways by resveratrol in chronic myelogenous leukemia (CML) cells results in the downregulation of Hsp70. PLoS One 5(1):e8719. Google Scholar
  2. Beher D, Wu J, Cumine S, Kim KW, Lu SC, Atangan L, Wang M (2009) Resveratrol is not a direct activator of SIRT1 enzyme activity. Chem Biol Drug Des 74(6):619–624. Google Scholar
  3. Benitez DA, Hermoso MA, Pozo-Guisado E, Fernández-Salguero PM, Castellón EA (2009) Regulation of cell survival by resveratrol involves inhibition of NFκB-regulated gene expression in prostate cancer cells. Prostate 69(10):1045–1054. Google Scholar
  4. Bickenbach KA, Veerapong J, Shao MY, Mauceri HJ, Posner MC, Kron SJ, Weichselbaum RR (2008) Resveratrol is an effective inducer of CArG-driven TNF-alpha gene therapy. Cancer Gene Ther 15(3):133–139. Google Scholar
  5. Borra MT, Smith BC, Denu JM (2005) Mechanism of human SIRT1 activation by resveratrol. J Biol Chem 280(17):17187–17195. Google Scholar
  6. Buhrmann C, Shayan P, Kraehe P, Popper B, Goel A, Shakibaei M (2015) Resveratrol induces chemosensitization to 5-fluorouracil through up-regulation of intercellular junctions, epithelial-to-mesenchymal transition and apoptosis in colorectal cancer. Biochem Pharmacol 98(1):51–68. Google Scholar
  7. Buhrmann C, Shayan P, Popper B, Goel A, Shakibaei M (2016) Sirt1 is required for resveratrol-mediated Chemopreventive effects in colorectal Cancer cells. Nutrients 8(3):145. Google Scholar
  8. Calderwood SK (2018) Heat shock proteins and cancer: intracellular chaperones or extracellular signalling ligands? Philos Trans R Soc Lond Ser B Biol Sci 373(1738):20160524. Google Scholar
  9. Catalgol B, Batirel S, Taga Y, Ozer NK (2012) Resveratrol: French paradox revisited. Front Pharmacol 3:141. Google Scholar
  10. Chai R, Fu H, Zheng Z, Liu T, Ji S, Li G (2017) Resveratrol inhibits proliferation and migration through SIRT1 mediated post-translational modification of PI3K/AKT signaling in hepatocellular carcinoma cells. Mol Med Rep 16(6):8037–8044. Google Scholar
  11. Chakraborty PK, Mustafi SB, Ganguly S, Chatterjee M, Raha S (2008) Resveratrol induces apoptosis in K562 (chronic myelogenous leukemia) cells by targeting a key survival protein, heat shock protein 70. Cancer Sci 99(6):1109–1116. Google Scholar
  12. Chan MM (2002) Antimicrobial effect of resveratrol on dermatophytes and bacterial pathogens of the skin. Biochem Pharmacol 63(2):99–104. Google Scholar
  13. Chatterjee S, Burns TF (2017) Targeting heat shock proteins in cancer: a promising therapeutic approach. Int J Mol Sci 18(9):E1978. Google Scholar
  14. Chen S, Xiao X, Feng X, Li W, Zhou N, Zheng L, Sun Y, Zhang Z, Zhu W (2012) Resveratrol induces Sirt1-dependent apoptosis in 3T3-L1 preadipocytes by activating AMPK and suppressing AKT activity and survivin expression. J Nutr Biochem 23(9):1100–1112. Google Scholar
  15. Chen X, Dong XS, Gao HY, Jiang YF, Jin YL, Chang YY, Chen LY, Wang JH (2016) Suppression of HSP27 increases the anti-tumor effects of quercetin in human leukemia U937 cells. Mol Med Rep 13(1):689–696. Google Scholar
  16. Concannon CG, Gorman AM, Samali A (2003) On the role of Hsp27 in regulating apoptosis. Apoptosis 8(1):61–70Google Scholar
  17. Dayalan Naidu S, Sutherland C, Zhang Y, Risco A, de la Vega L, Caunt CJ, Hastie CJ, Lamont DJ, Torrente L, Chowdhry S, Benjamin IJ, Keyse SM, Cuenda A, Dinkova-Kostova AT (2016) Heat shock factor 1 is a substrate for p38 mitogen-activated protein kinases. Mol Cell Biol 36(18):2403–2417. Google Scholar
  18. Del Follo-Martinez A, Banerjee N, Li X, Safe S, Mertens-Talcott S (2013) Resveratrol and quercetin in combination have anticancer activity in colon cancer cells and repress oncogenic microRNA-27a. Nutr Cancer 65(3):494–504. Google Scholar
  19. Delmas D, Rebe C, Lacour S, Filomenko R, Athias A, Gambert P, Cherkaoui-Malki M, Jannin B, Dubrez-Daloz L, Latruffe N, Solary E (2003) Resveratrol-induced apoptosis is associated with Fas redistribution in the rafts and the formation of a death-inducing signaling complex in colon cancer cells. J Boil Chem 278(42):41482–41490. Google Scholar
  20. Díaz-Chávez J, Fonseca-Sánchez MA, Arechaga-Ocampo E, Flores-Pérez A, Palacios-Rodríguez Y, Domínguez-Gómez G, Marchat LA, Fuentes-Mera L, Mendoza-Hernández G, Gariglio P, López-Camarillo C (2013) Proteomic profiling reveals that resveratrol inhibits HSP27 expression and sensitizes breast cancer cells to doxorubicin therapy. PLoS One 8(5):e64378. Google Scholar
  21. Docherty JJ, Fu MM, Stiffler BS, Limperos RJ, Pokabla CM, DeLucia AL (1999) Resveratrol inhibition of herpes simplex virus replication. Antivir Res 43(3):145–155. Google Scholar
  22. Elbashir SM, Harborth J, Lendeckel W, Yalcin A, Weber K, Tuschl T (2001) Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 411(6836):494–498. Google Scholar
  23. Elshaer M, Chen Y, Wang XJ, Tang X (2018) Resveratrol: an overview of its anti-cancer mechanisms. Life Sci 207:340–349. Google Scholar
  24. Frazzi R, Valli R, Tamagnini I, Casali B, Latruffe N, Merli F (2013) Resveratrol-mediated apoptosis of hodgkin lymphoma cells involves SIRT1 inhibition and FOXO3a hyperacetylation. Int J Cancer 132(5):1013–1021. Google Scholar
  25. Ganapathy S, Chen Q, Singh KP, Shankar S, Srivastava RK (2010) Resveratrol enhances antitumor activity of TRAIL in prostate cancer xenografts through activation of FOXO transcription factor. PLoS One 5(12):e15627. Google Scholar
  26. Garrido C, Brunet M, Didelot C, Zermati Y, Schmitt E, Kroemer G (2006) Heat shock proteins 27 and 70: anti-apoptotic proteins with tumourigenic properties. Cell Cycle 5(22):2592–2601. Google Scholar
  27. He X, Wang Y, Zhu J, Orloff M, Eng C (2011) Resveratrol enhances the anti-tumor activity of the mTOR inhibitor rapamycin in multiple breast cancer cell lines mainly by suppressing rapamycin-induced AKT signaling. Cancer Lett 301(2):168–176. Google Scholar
  28. Holmes B, Benavides-Serrato A, Freeman RS, Landon KA, Bashir T, Nishimura RN, Gera J (2018) mTORC2/AKT/HSF1/HuR constitute a feed-forward loop regulating Rictor expression and tumor growth in glioblastoma. Oncogene 37(6):732–743. Google Scholar
  29. Huang J, Gan Q, Han L, Li J, Zhang H, Sun Y, Zhang Z, Tong T (2008) SIRT1 overexpression antagonizes cellular senescence with activated ERK/S6K1 signaling in human diploid fibroblasts. PLoS One 3(3):e1710. Google Scholar
  30. Jakubowicz-Gil J, Langner E, Bądziul D, Wertel I, Rzeski W (2013a) Apoptosis induction in human glioblastoma multiforme T98G cells upon temozolomide and quercetin treatment. Tumour Biol 34(4):2367–2378. Google Scholar
  31. Jakubowicz-Gil J, Langner E, Bądziul D, Wertel I, Rzeski W (2013b) Silencing of Hsp27 and Hsp72 in glioma cells as a tool for programmed cell death induction upon temozolomide and quercetin treatment. Toxicol Appl Pharmacol 273(3):580–589. Google Scholar
  32. Jakubowicz-Gil J, Langner E, Wertel I, Piersiak T, Rzeski W (2010) Temozolomide, quercetin and cell death in the MOGGCCM astrocytoma cell line. Chem Biol Interact 188(1):190–203. Google Scholar
  33. Jiang H, Shang X, Wu H, Gautam SC, Al-Holou S, Li C, Kuo J, Zhang L, Chopp M (2009) Resveratrol downregulates PI3K/Akt/mTOR signaling pathways in human U251 glioma cells. J Exp Ther Oncol 8(1):25–33Google Scholar
  34. Juan ME, Wenzel U, Daniel H, Planas JM (2008) Resveratrol induces apoptosis through ROS-dependent mitochondria pathway in HT-29 human colorectal carcinoma cells. J Agric Food Chem 56(12):4813–4818. Google Scholar
  35. Khalil AA (2007) Biomarker discovery: a proteomic approach for brain cancer profiling. Cancer Sci 98(2):201–213. Google Scholar
  36. Khalil AA, Kabapy NF, Deraz SF, Smith C (2011) Heat shock proteins in oncology: diagnostic biomarkers or therapeutic targets? Biochim Biophys Acta 1816(2):89–104. Google Scholar
  37. Khan F, Niaz K, Maqbool F, Ismail Hassan F, Abdollahi M, Nagulapalli Venkata KC, Nabavi SM, Bishayee A (2016) Molecular targets underlying the anticancer effects of quercetin: an update. Nutrients 8(9):529–548. Google Scholar
  38. Kim YA, Lee WH, Choi TH, Rhee SH, Park KY, Choi YH (2003) Involvement of p21WAF1/CIP1, pRB, Bax and NF-κB in induction of growth arrest and apoptosis by resveratrol in human lung carcinoma A549 cells. Int J Oncol 23(4):1143–1149Google Scholar
  39. Ko JH, Sethi G, Um JY, Shanmugam MK, Arfuso F, Kumar AP, Bishayee A, Ahn KS (2017) The role of resveratrol in cancer therapy. Int J Mol Sci 18(12):E2589. Google Scholar
  40. Li J, Tang C, Li L, Li R, Fan Y (2016) Quercetin blocks t-AUCB-induced autophagy by Hsp27 and Atg7 inhibition in glioblastoma cells in vitro. J Neuro-Oncol 129(1):39–45. Google Scholar
  41. Li L, Qiu RL, Lin Y, Cai Y, Bian Y, Fan Y, Gao XJ (2018) Resveratrol suppresses human cervical carcinoma cell proliferation and elevates apoptosis via the mitochondrial and p53 signaling pathways. Oncol Lett 15(6):9845–9851. Google Scholar
  42. Lin CJ, Lee CC, Shih YL, Lin TY, Wang SH, Lin YF, Shih CM (2012) Resveratrol enhances the therapeutic effect of temozolomide against malignant glioma in vitro and in vivo by inhibiting autophagy. Free Radic Biol Med 52(2):377–391. Google Scholar
  43. Narayanan NK, Nargi D, Randolph C, Narayanan BA (2009) Liposome encapsulation of curcumin and resveratrol in combination reduces prostate cancer incidence in PTEN knockout mice. Int J Cancer 125(1):1–8. Google Scholar
  44. Nylandsted J, Rohde M, Brand K, Bastholm L, Elling F, Jäättelä M (2000) Selective depletion of heat shock protein 70 (Hsp70) activates a tumor-specific death program that is independent of caspases and bypasses Bcl-2. Proc Natl Acad Sci U S A 97(14):7871–7876Google Scholar
  45. Önay Uçar E, Pekmez M, Arda N (2017) Targeting of heat shock proteins by natural products in cancer. In: Farooqi A, Ismail M (eds) Molecular oncology: underlying mechanisms and translational advancements. Springer, Cham, pp 173–192 ISBN: 978-3-319-53081-9. Google Scholar
  46. Önay Uçar E, Şengelen A, Mertoğlu E, Pekmez M, Arda N (2018) Suppression of HSP70 expression by quercetin and its therapeutic potential against cancer. In: Asea A, Kaur P (eds) HSP70 in Human diseases and disorders. heat shock proteins, 14th edn. Springer, Cham, pp 361–379 ISBN: 978–3–319-89550-5. doi:
  47. Önay-Uçar E (2015) Heat shock proteins and cancer: plant based therapy. In: Asea AAA, Calderwoo SK (eds) Heat shock protein-based therapies, 9th edn. Springer, Cham, pp 27–48 ISBN: 978–3–319-17210-1. Google Scholar
  48. Önay-Uçar E, Şengelen A, Güngör E, Mertoğlu E, Pekmez M, Arda N (in press) Can hsp targeted gene therapy be a new hope for gliomas? In: Asea A, Kaur P (eds) Heat shock proteins in neuroscience, 20th edn. SpringerGoogle Scholar
  49. Parekh P, Motiwale L, Naik N, Rao KV (2011) Downregulation of cyclin D1 is associated with decreased levels of p38 MAP kinases, Akt/PKB and Pak1 during chemopreventive effects of resveratrol in liver cancer cell. Exp Toxicol Pathol 63(1–2):167–173. Google Scholar
  50. Pervaiz S, Holme AL (2009) Resveratrol: its biologic targets and functional activity. Antioxid Redox Signal 11(11):2851–2897. Google Scholar
  51. Rajesh Y, Pal I, Banik P, Chakraborty S, Borkar SA, Dey G, Mukherjee A, Mandal M (2017) Insights into molecular therapy of glioma: current challenges and next generation blueprint. Acta Pharmacol Sin 38(5):591–613. Google Scholar
  52. Ren Z, Wang L, Cui J, Huoc Z, Xue J, Cui H, Mao Q, Yang R (2013) Resveratrol inhibits NF-kB signaling through suppression of p65 and IkappaB kinase activities. Pharmazie 68(8):689–694Google Scholar
  53. Rocha-González HI, Ambriz-Tututi M, Granados-Soto V (2008) Resveratrol: a natural compound with pharmacological potential in neurodegenerative diseases. CNS Neurosci Ther 14(3):234–247. Google Scholar
  54. Rubiolo JA, Vega FV (2008) Resveratrol protects primary rat hepatocytes against necrosis induced by reactive oxygen species. Biomed Pharmacother 62(9):606–612. Google Scholar
  55. Şengelen A, Önay-Uçar E (2018) Rosmarinic acid and siRNA combined therapy represses Hsp27 (HSPB1) expression and induces apoptosis in human glioma cells. Cell Stress Chaperones 23(5):885–896. Google Scholar
  56. Sengottuvelan M, Deeptha K, Nalini N (2009) Influence of dietary resveratrol on early and late molecular markers of 1,2-dimethylhydrazine-induced colon carcinogenesis. Nutrition 25(11–12):1169–1176. Google Scholar
  57. Shim G, Kim D, Le QV, Park GT, Kwon T, Oh YK (2018) Nonviral delivery systems for cancer gene therapy: strategies and challenges. Curr Gene Ther 18(1):3–20. Google Scholar
  58. Shimizu T, Nakazato T, Xian MJ, Sagawa M, Ikeda Y, Kizaki M (2006) Resveratrol induces apoptosis of human malignant B cells by activation of caspase-3 and p38 MAP kinase pathways. Biochem Pharmacol 71(6):742–750. Google Scholar
  59. Staedler D, Idrizi E, Kenzaoui BH, Juillerat-Jeanneret L (2011) Drug combinations with quercetin: doxorubicin plus quercetin in human breast cancer cells. Cancer Chemother Pharmacol 68(5):1161–1172. Google Scholar
  60. Uçar EÖ, Arda N, Aitken A (2012) Extract from mistletoe, Viscum album L., reduces Hsp27 and 14-3-3 protein expression and induces apoptosis in C6 rat glioma cells. Genet Mol Res 11(3):2801–2813. Google Scholar
  61. Wang G, Dai F, Yu K, Jia Z, Zhang A, Huang Q, Kang C, Jiang H, Pu P (2015) Resveratrol inhibits glioma cell growth via targeting oncogenic microRNAs and multiple signaling pathways. Int J Oncol 46(4):1739–1747Google Scholar
  62. Wang X, Khaleque MA, Zhao MJ, Zhong R, Gaestel M, Calderwood SK (2006) Phosphorylation of HSF1 by MAPK-activated protein kinase 2 on serine 121, inhibits transcriptional activity and promotes HSP90 binding. J Biol Chem 281(12):782–791. Google Scholar
  63. Wang Z, Raoi DD, Senzer N, Nemunaitis J (2011) RNA interference and cancer therapy. Pharm Res 28(12):2983–2995. Google Scholar
  64. Westerheide SD, Anckar J, Stevens SM Jr, Sistonen L, Morimoto RI (2009) Stress-inducible regulation of heat shock factor 1 by the deacetylase SIRT1. Science 323(5917):1063–1066. Google Scholar
  65. Westphal M, Lamszus K (2011) The neurobiology of gliomas: from cell biology to the development of therapeutic approaches. Nat Rev Neurosci 12(9):495–508. Google Scholar
  66. Yang Q, Wang B, Zang W, Wang X, Liu Z, Li W, Jia J (2013) Resveratrol inhibits the growth of gastric cancer by inducing G1 phase arrest and senescence in a Sirt1-dependent manner. PLoS One 8(11):e70627. Google Scholar
  67. Zhang L, Guo X, Xie W, Li Y, Ma M, Yuan T, Luo B (2015) Resveratrol exerts an anti-apoptotic effect on human bronchial epithelial cells undergoing cigarette smoke exposure. Mol Med Rep 11(3):1752–1758. Google Scholar
  68. Zhang R, Tremblay TL, McDermid A, Thibault P, Stanimirovic D (2003) Identification of differentially expressed proteins in human glioblastoma cell lines and tumors. Glia 42(2):194–208. Google Scholar
  69. Zhao Y, Tang H, Zeng X, Ye D, Liu J (2018) Resveratrol inhibits proliferation, migration and invasion via Akt and ERK1/2 signaling pathways in renal cell carcinoma cells. Biomed Pharmacother 98:36–44. Google Scholar
  70. Zhou F, Huang X, Pan Y, Cao D, Liu C, Liu Y, Chen A (2018) Resveratrol protects HaCaT cells from ultraviolet B-induced photoaging via upregulation of HSP27 and modulation of mitochondrial caspase-dependent apoptotic pathway. Biochem Biophys Res Commun 499(3):662–668. Google Scholar

Copyright information

© Cell Stress Society International 2019

Authors and Affiliations

  1. 1.Department of Molecular Biology and Genetics, Faculty of ScienceIstanbul UniversityIstanbulTurkey
  2. 2.Department of Molecular Biology and Genetics, Institute of Graduate Studies in SciencesIstanbul UniversityIstanbulTurkey

Personalised recommendations