Dendrosomal nanocurcumin and p53 overexpression synergistically trigger apoptosis in glioblastoma cells

Document Type : Original Article

Authors

Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran

Abstract

Objective(s): Glioblastoma is the most lethal tumor of the central nervous system. Here, we aimed to evaluate the effects of exogenous delivery of p53 and a nanoformulation of curcumin called dendrosomal curcumin (DNC), alone and in combination, on glioblastoma tumor cells.
Materials and Methods: MTT assay was exploited to measure the viability of U87-MG cells against DNC treatment. Cells were separately subjected to DNC treatment and transfected with p53-containing vector and then were co-exposed to DNC and p53 overexpression. Annexin-V-FLUOS staining followed by flow cytometry and real-time PCR were applied to examine apoptosis and analyze the expression levels of the genes involved in cell cycle and oncogenesis, respectively.
Results: The results of cell viability assay through MTT indicated that DNC inhibits the proliferation of U87-MG cells in a time- and dose-dependent manner. Apoptosis evaluation revealed that p53 overexpression accompanied by DNC treatment can act in a synergistic manner to significantly enhance the number of apoptotic cells (90%) compared with their application alone (15% and 38% for p53 overexpression and DNC, respectively). Also, real-time PCR data showed that the concomitant exposure of cells to both DNC and p53 overexpression leads to an enhanced expression of GADD45 and a reduced expression of NF-κB and c-Myc.
Conclusion: The findings of the current study suggest that our combination strategy, which merges two detached gene (p53) and drug (curcumin) delivery systems into an integrated platform, may represent huge potential as a novel and efficient modality for glioblastoma treatment.

Keywords


1. Meng J, Li P, Zhang Q, Yang Z, Fu S. A radiosensitivity gene signature in predicting glioma prognostic via EMT pathway. Oncotarget 2014; 5:4683-4693.
2. Chinot OL, Wick W, Mason W, Henriksson R, Saran F, Nishikawa R, et al. Bevacizumab plus radiotherapy-temozolomide for newly diagnosed glioblastoma. N Engl J  Med 2014;370:709-722.
3. Sa JK, Yoon Y, Kim M, Kim Y, Cho HJ, Lee JK, et al. In vivo RNAi screen identifies NLK as a negative regulator of mesenchymal activity in glioblastoma. Oncotarget 2015; 6:20145-59.
4. Tang Y, Zhao W, Chen Y, Zhao Y, Gu W. Acetylation is indispensable for p53 activation. Cell 2008; 133:612-626.
5. Badie B, Goh CS, Klaver J, Herweijer H, Boothman DA. Combined radiation and p53 gene therapy of malignant glioma cells. Cancer Gene Ther 1999; 6:155-162.
6. Peller S, Rotter V. TP53 in hematological cancer: low incidence of mutations with significant clinical relevance. Hum Mutat 2003; 21:277-284.
7. Blons H, Laurent-Puig P. TP53 and head and neck neoplasms. Hum Mutat 2003;21:252-257.
8. Iacopetta B. TP53 mutation in colorectal cancer. Hum Mutat 2003; 21:271-276.
9. Schuijer M, Berns EM. TP53 and ovarian cancer. Hum Mutat 2003; 21:285-291.
10. Sun Y. E3 ubiquitin ligases as cancer targets and biomarkers. Neoplasia 2006; 8:645-654.
11. Lee SJ, Krauthauser C, Maduskuie V, Fawcett PT, Olson JM, Rajasekaran SA. Curcumin-induced HDAC inhibition and attenuation of medulloblastoma growth in vitro and in vivo. BMC Cancer 2011; 11:144.
12. Choi BH, Kim CG, Bae YS, Lim Y, Lee YH, Shin SY. p21 Waf1/Cip1 expression by curcumin in U-87MG human glioma cells: role of early growth response-1 expression. Cancer Res 2008; 68:1369-1377.
13. Alizadeh AM, Sadeghizadeh M, Najafi F, Ardestani SK, Erfani-Moghadam V, Khaniki M, et al. Encapsulation of curcumin in diblock copolymer micelles for cancer therapy. Biomed Res Int 2015; 2015:824746.
14. Kunnumakkara AB, Anand P, Aggarwal BB. Curcumin inhibits proliferation, invasion, angiogenesis and metastasis of different cancers through interaction with multiple cell signaling proteins. Cancer Lett 2008; 269:199-225.
15. Dudas J, Fullar A, Romani A, Pritz C, Kovalszky I, Hans Schartinger V, et al. Curcumin targets fibroblast-tumor cell interactions in oral squamous cell carcinoma. Exp Cell Res 2013; 319:800-809.
16. Tahmasebi Mirgani M, Isacchi B, Sadeghizadeh M, Marra F, Bilia AR, Mowla SJ, et al. Dendrosomal curcumin nanoformulation downregulates pluripotency genes via miR-145 activation in U87MG glioblastoma cells. Int J Nanomed 2014; 9:403-417.
17. Orr WS, Denbo JW, Saab KR, Myers AL, Ng CY, Zhou J, et al. Liposome-encapsulated curcumin suppresses neuroblastoma growth through nuclear factor-kappa B inhibition. Surgery 2012; 151:736-744.
18. Bisht S, Feldmann G, Soni S, Ravi R, Karikar C, Maitra A, et al. Polymeric nanoparticle-encapsulated curcumin ("nanocurcumin"): a novel strategy for human cancer therapy. J Nanobiotechnol 2007; 5:3.
19. Babaei E, Sadeghizadeh M, Hassan ZM, Feizi MA, Najafi F, Hashemi SM. Dendrosomal curcumin significantly suppresses cancer cell proliferation in vitro and in vivo. Int Immunopharmacol 2012; 12:226-234.
20. Sadeghizadeh M, Ranjbar B, Damaghi M, Khaki L, Sarbolouki MN, Najafi F, et al. Dendrosomes as novel gene porters‐III. J Chem Technol Biotechnol 2008; 83:912-920.
21. Sarbolouki MN, Sadeghizadeh M, Yaghoobi MM, Karami A, Lohrasbi T. Dendrosomes: a novel family of vehicles for transfection and therapy. J Chem Technol Biotechnol 2000; 75:919-922.
22. Mirgani MT, Isacchi B, Sadeghizadeh M, Marra F, Bilia AR, Mowla SJ, et al. Dendrosomal curcumin nanoformulation downregulates pluripotency genes via miR-145 activation in U87MG glioblastoma cells. Int J Nanomed 2014; 9:403-417.
23. Alizadeh AM, Khaniki M, Azizian S, Mohaghgheghi MA, Sadeghizadeh M, Najafi F. Chemoprevention of azoxymethane-initiated colon cancer in rat by using a novel polymeric nanocarrier--curcumin. Eur J Pharmacol 2012; 689:226-232.
24. Farhangi B, Alizadeh AM, Khodayari H, Khodayari S, Dehghan MJ, Khori V, et al. Protective effects of dendrosomal curcumin on an animal metastatic breast tumor. Eur J Pharmacol 2015; 758:188-196.
25. Mohajeri M, Sadeghizadeh M, Najafi F, Javan M. Polymerized nano-curcumin attenuates neurological symptoms in EAE model of multiple sclerosis through down regulation of inflammatory and oxidative processes and enhancing neuroprotection and myelin repair. Neuropharmacology 2015; 99:156-167.
26. Gou M, Men K, Shi H, Xiang M, Zhang J, Song J, et al. Curcumin-loaded biodegradable polymeric micelles for colon cancer therapy in vitro and in vivo. Nanoscale 2011; 3:1558-1567.
27. Lai Lh, Fu Qh, Liu Y, Jiang K, Guo Qm, Chen Qy, et al. Piperine suppresses tumor growth and metastasis in vitro and in vivo in a 4T1 murine breast cancer model. Acta Pharmacol Sin 2012; 33:523-5230.
28. Massumi M, Ziaee AA, Sarbolouki MN. Apoptosis induction in human lymphoma and leukemia cell lines by transfection via dendrosomes carrying wild-type p53 cDNA. Biotechnol Lett 2006;28(1):61-6.
29. Dehghan Esmatabadi MJ, Farhangi B, Safari Z, Kazerooni H, Shirzad H, Zolghadr F, et al. Dendrosomal curcumin inhibits metastatic potential of human SW480 colon cancer cells through Down-regulation of Claudin1, Zeb1 and Hef1-1 gene expression. Asian Pac J Cancer Prev 2015; 16:2473-2481.
30. Sun Y. p53 and its downstream proteins as molecular targets of cancer. Mol Carcinog 2006; 45:409-415.
31. Fridman JS, Lowe SW. Control of apoptosis by p53. Oncogene 2003; 22:9030-9040.
32. Amaral JD, Xavier JM, Steer CJ, Rodrigues CM. The role of p53 in apoptosis. Discov Med 2010; 9:145-152.
33. Benchimol S. p53-dependent pathways of apoptosis. Cell Death Differ 2001; 8:1049-10451.
34. Khaw AK, Hande MP, Kalthur G, Hande MP. Curcumin inhibits telomerase and induces telomere shortening and apoptosis in brain tumour cells. J Cell Bbiochem 2013; 114:1257-1270.
35. Noorafshan A, Ashkani-Esfahani S. A review of therapeutic effects of curcumin. Curr Pharm Des 2013; 19:2032-2046.
36. Reuter S, Eifes S, Dicato M, Aggarwal BB, Diederich M. Modulation of anti-apoptotic and survival pathways by curcumin as a strategy to induce apoptosis in cancer cells. Biochem Pharmacol 2008; 76:1340-1351.
37. Jee SH, Shen SC, Kuo ML, Tseng CR, Chiu HC. Curcumin induces a p53-dependent apoptosis in human basal cell carcinoma cells.J Invest Dermatol 1998; 111:656-661.
38. Gomez-Manzano C, Fueyo J, Kyritsis AP, Steck PA, Roth JA, McDonnell TJ, et al. Adenovirus-mediated transfer of the p53 gene produces rapid and generalized death of human glioma cells via apoptosis. Cancer Res 1996; 56:694-699.
39. Sun Y. E3 ubiquitin ligases as cancer targets and biomarkers. Neoplasia 2006; 8:645-654.
40. Goel A, Kunnumakkara AB, Aggarwal BB. 
Curcumin as “Curecumin”: from kitchen to clinic. Biochem Pharmacol 2008; 75:787-809.
41. Zerbini LF, Libermann TA. Life and death in cancer GADD45 α and γ are critical regulators of NF-κB mediated escape from programmed cell death. Cell Cycle 2005; 4:18-20.
42. Bales KR, Du Y, Dodel RC, Yan GM, Hamilton-Byrd E, Paul SM. The NF-κB/Rel family of proteins mediates Aβ-induced neurotoxicity and glial activation. Brain Res Mol Brain Res 1998; 57:63-72.
43. Guo Q, Robinson N, Mattson MP. Secreted β-amyloid precursor protein counteracts the proapoptotic action of mutant presenilin-1 by activation of NF-κB and stabilization of calcium homeostasis. J Biol Chem 1998; 273:12341-12351.
44. Gerondakis S, Grossmann M, Nakamura Y, Pohl T, Grumont R. Genetic approaches in mice to understand Rel/NF-kappaB and IkappaB function: transgenics and knockouts. Oncogene 1999; 18:6888-6895.
45. Mattson MP, Camandola S. NF-κB in neuronal plasticity and neurodegenerative disorders. J Clin Invest 2001; 107:247-254.
46. Chen F, Castranova V, Shi X. New insights into the role of nuclear factor-κB in cell growth regulation. Am J Pathol 2001; 159:387-397.
47. Kastan MB, Zhan Q, El-Deiry WS, Carrier F, Jacks T, Walsh WV, et al. A mammalian cell cycle checkpoint pathway utilizing p53 and GADD45 is defective in ataxia-telangiectasia. Cell 1992; 71:587-597.
48. Salvador JM, Brown-Clay JD, Fornace Jr AJ. Gadd45 in stress signaling, cell cycle control, and apoptosis. Adv Exp Med Biol 2013; 793:1-19.
49. Zerbini LF, Wang Y, Correa RG, Cho JY, Libermann TA. Blockage of NF-kappaB induces serine 15 phosphorylation of mutant p53 by JNK kinase in prostate cancer cells. Cell Cycle 2005; 4:1247-1253.
50. Hildesheim J, Bulavin DV, Anver MR, Alvord WG, Hollander MC, Vardanian L, et al. Gadd45a protects against UV irradiation-induced skin tumors, and promotes apoptosis and stress signaling via MAPK and p53. Cancer Res 2002; 62:7305-7315.
51. Tong T, Ji J, Jin S, Li X, Fan W, Song Y, et al. Gadd45a expression induces Bim dissociation from the cytoskeleton and translocation to mitochondria. Mol Cell Biol 2005; 25:4488-4500.