Effects of 8-hydroxyquinoline-coated graphene oxide on cell death and apoptosis in MCF-7 and MCF-10 breast cell lines

Document Type: Original Article


1 Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran

2 Cancer Immunotherapy and Regenerative Medicine, Breast Cancer Research Center, Motamed Cancer Institute, ACECR, Tehran, Iran

3 School of Chemistry, College of Science, University of Tehran, Tehran, Iran


Objective(s): Breast cancer is a devastating disease related to women. The anticancer properties of 8-hydroxyquinoline (8HQ) and the increasing use of graphene oxide (GO), as a drug delivery system with anti-cancerous properties, led us to investigate the toxicity and apoptosis-induction capability of 8HQ-coated GO on breast cancer cells compared with normal breast cells.
Materials and Methods: Breast cancer (MCF-7) and normal breast (MCF-10) cell lines were treated with several doses of 8-HQ-coated GO for 12, 24, and 48 hr. The toxicity of the nanocomposite was measured using MTT assay and the effect of the nanocomposite on cell apoptosis was determined by examining the expression of P53, P21, Bax, and BCL2 genes, as well as Annexin-V /PI apoptosis assay.
Results: There were significantly increased cell deaths in nanocomposite-treated MCF-7 breast cancer cells, compared with treated normal breast cells. Significantly increased expression of P53, P21, and Bax genes and reduced expression of BCL2 gene were found in the treated breast cancer cell line compared with the normal cell line. Annexin-V/PI assay also illustrated significant induction of apoptosis in MCF-7 following nanocomposite treatment.
Conclusion: Overall, 8HQ-coated GO has toxicity for breast cancer cell lines and one of the mechanisms through which this nanocomposite can induce cell death is the induction of apoptosis. With complementary  in vitro and in vivo studies, this nanocomposite can be suggested as a nano-drug with anti-cancer properties.


1. Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA cancer J Clin 2018; 68:394-424.
2. Sharma GN, Dave R, Sanadya J, Sharma P, Sharma K. Various types and management of breast cancer: an overview. J Adv Pharm Technol Res 2010; 1:109-126.
3. Stewart BW. Mechanisms of apoptosis: integration of genetic, biochemical, and cellular indicators. J Natl Cancer Inst 1994; 86:1286-1296.
4. Dakubo GD. Mitochondrial genetics and cancer: Springer Science & Business Media; 2010.
5.El-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, et al. WAF1, a potential mediator of p53 tumor suppression. Cell 1993; 75:817-825.
6. Bunz F, Dutriaux A, Lengauer C, Waldman T, Zhou S, Brown J, et al. Requirement for p53 and p21 to sustain G2 arrest after DNA damage. Science 1998; 282:1497-1501.
7. Fulda S, Debatin K-M. Extrinsic versus intrinsic apoptosis pathways in anticancer chemotherapy. Oncogene 2006; 25:4798.
8. Pierson HO. Handbook of carbon, graphite, diamonds and fullerenes: processing, properties and applications: William Andrew; 2012.
9. Du W, Jiang X, Zhu L. From graphite to graphene: direct liquid-phase exfoliation of graphite to produce single-and few-layered pristine graphene. J Mater Chem A Mater 2013; 1:10592-10606.
10. Paulchamy B, Arthi G, Lignesh B. A simple approach to stepwise synthesis of graphene oxide nanomaterial. J Nanomed Nanotechnol 2015; 6:253-256.
11. Wei J, Vo T, Inam F. Epoxy/graphene nanocomposites–processing and properties: A review. RSC Adv 2015; 5:73510-73524.
12. Srivastava V, Negi AS, Kumar J, Gupta M, Khanuja SP. Plant-based anticancer molecules: a chemical and biological profile of some important leads. Bioorg Med Chem 2005; 13:5892-5908.
13. Afzal O, Kumar S, Haider MR, Ali MR, Kumar R, Jaggi M, et al. A review on anticancer potential of bioactive heterocycle quinoline. Eur J Med Chem 2015; 97:871-910.
14. Xu H, Chen W, Zhan P, Liu X. 8-Hydroxyquinoline: a privileged structure with a broad-ranging pharmacological potential. Med Chem Comm 2015; 6:61-74.
15. Bianco A. Graphene: safe or toxic? The two faces of the medal. Angew Chem Int Ed Engl 2013; 52:4986-4997.
16. Hummers Jr WS, Offeman RE. Preparation of graphitic oxide. J Am Chem Soc 1958; 80:1339-1339.
17. Badiei A, Goldooz H, Ziarani GM. A novel method for preparation of 8-hydroxyquinoline functionalized mesoporous silica: Aluminum complexes and photoluminescence studies. Appl Surf Sci 2011; 257:4912-4918.
18. Goldooz H, Badiei A, Shiravand G, Ghasemi JB, Ziarani GM. A highly selective Ag+ sensor based on 8-hydroxyquinoline functionalized graphene oxide-silica nanosheet and its logic gate behaviour. J Mater Sci 2019; 30:17693-17705.
19. Priyadarsini RV, Murugan RS, Maitreyi S, Ramalingam K, Karunagaran D, Nagini S. The flavonoid quercetin induces cell cycle arrest and mitochondria-mediated apoptosis in human cervical cancer (HeLa) cells through p53 induction and NF-κB inhibition. Eur J Pharmacol 2010; 649:84-91.
20. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. methods 2001; 25:402-408.
21. Lei H, Xie M, Zhao Y, Zhang F, Xu Y, Xie J. Chitosan/sodium alginate modificated graphene oxide-based nanocomposite as a carrier for drug delivery. Ceram Int 2016; 42:17798-17805.
22. Liu Z, Robinson JT, Sun X, Dai H. PEGylated nanographene oxide for delivery of water-insoluble cancer drugs. J AM Chem Soc 2008; 130:10876-10877.
23. Yadav N, Kumar N, Prasad P, Shirbhate S, Sehrawat S, Lochab B. Stable dispersions of covalently tethered polymer improved graphene oxide nanoconjugates as an effective vector for siRNA delivery. ACS Appl Mater Interfaces 2018; 10:14577-14593.
24. Chang Y, Yang S-T, Liu J-H, Dong E, Wang Y, Cao A, et al. In vitro toxicity evaluation of graphene oxide on A549 cells. Toxicol Lett 2011; 200:201-210.
25. Chen L, Hu P, Zhang L, Huang S, Luo L, Huang C. Toxicity of graphene oxide and multi-walled carbon nanotubes against human cells and zebrafish. Sci China Chem 2012; 55:2209-2216.
26. Alibolandi M, Mohammadi M, Taghdisi SM, Ramezani M, Abnous K. Fabrication of aptamer decorated dextran coated nano-graphene oxide for targeted drug delivery. Carbohydr Polym 2017; 155:218-229.
27. Liu Y, Zhong H, Qin Y, Zhang Y, Liu X, Zhang T. Non-covalent hydrophilization of reduced graphene oxide used as a paclitaxel vehicle. RSC Adv 2016; 6:30184-30193.
28. Chaudhari NS, Pandey AP, Patil PO, Tekade AR, Bari SB, Deshmukh PK. Graphene oxide based magnetic nanocomposites for efficient treatment of breast cancer. Mater Sci Eng C 2014; 37:278-285.
29. Hu W, Peng C, Luo W, Lv M, Li X, Li D, et al. Graphene-based antibacterial paper. ACS Nano 2010; 4:4317-4323.
30. Krajnović T, Maksimović-Ivanić D, Mijatović S, Drača D, Wolf K, Edeler D, et al. Drug delivery system for emodin based on mesoporous silica SBA-15. Nanomaterials 2018; 8:322-337.
31. Wang D, Huang J, Wang X, Yu Y, Zhang H, Chen Y, et al. The eradication of breast cancer cells and stem cells by 8-hydroxyquinoline-loaded hyaluronan modified mesoporous silica nanoparticle-supported lipid bilayers containing docetaxel. Biomaterials 2013; 34:7662-7673.
32. Dreaden EC, Alkilany AM, Huang X, Murphy CJ, El-Sayed MA. The golden age: gold nanoparticles for biomedicine. Chem Soc Rev 2012; 41:2740-2779.
33. Liu W, Li X, Wong Y-S, Zheng W, Zhang Y, Cao W, et al. Selenium nanoparticles as a carrier of 5-fluorouracil to achieve anticancer synergism. Acs Nano 2012; 6:6578-6591.
34. Lv Y, Tao L, Bligh SA, Yang H, Pan Q, Zhu L. Targeted delivery and controlled release of doxorubicin into cancer cells using a multifunctional graphene oxide. Mater Sci Eng C 2016; 59:652-660.
35. Jafarizad A, Aghanejad A, Sevim M, Metin Ö, Barar J, Omidi Y, et al. Gold nanoparticles and reduced graphene oxide‐gold nanoparticle composite materials as covalent drug delivery systems for breast cancer treatment. ChemistrySelect 2017; 2:6663-6672.
36. Fan L, Ge H, Zou S, Xiao Y, Wen H, Li Y, et al. Sodium alginate conjugated graphene oxide as a new carrier for drug delivery system. Int J Biol Macromol 2016; 93:582-590.
37. Fiorillo M, Verre AF, Iliut M, Peiris-Pagés M, Ozsvari B, Gandara R, et al. Graphene oxide selectively targets cancer stem cells, across multiple tumor types: Implications for non-toxic cancer treatment, via “differentiation-based nano-therapy”. Oncotarget 2015; 6:3553-3562.
38. Yoon HH, Bhang SH, Kim T, Yu T, Hyeon T, Kim BS. Dual roles of graphene oxide in chondrogenic differentiation of adult stem cells: Cell‐adhesion substrate and growth factor‐delivery carrier. Adv Func Mater 2014; 24:6455-6464.
39. Prachayasittikul V, Prachayasittikul S, Ruchirawat S, Prachayasittikul V. 8-Hydroxyquinolines: a review of their metal chelating properties and medicinal applications. Drug Des Devel Ther 2013; 7:1157.
40. Zhai S, Yang L, Cui QC, Sun Y, Dou QP, Yan B. Tumor cellular proteasome inhibition and growth suppression by 8-hydroxyquinoline and clioquinol requires their capabilities to bind copper and transport copper into cells. J Biol Inorg Chemi 2010; 15:259-269.
41. Wang N, Świtalska M, Wu MY, Imai K, Ngoc TA, Pang CQ, et al. Synthesis and in vitro cytotoxic effect of 6-amino-substituted 11H-and 11Me-indolo [3, 2-c] quinolines. Eur J Med Chem 2014; 78:314-323.
42. Xiao Z, Lei F, Chen X, Wang X, Cao L, Ye K, et al. Design, synthesis, and antitumor evaluation of quinoline‐imidazole derivatives. Arch Pharm 2018; 351:1700407.
43. Chen C, Hou X, Wang G, Pan W, Yang X, Zhang Y, et al. Design, synthesis and biological evaluation of quinoline derivatives as HDAC class I inhibitors. Eur J Med Chem 2017; 133:11-23.
44. Qin Q-P, Chen Z-F, Qin J-L, He X-J, Li Y-L, Liu Y-C, et al. Studies on antitumor mechanism of two planar platinum (II) complexes with 8-hydroxyquinoline: synthesis, characterization, cytotoxicity, cell cycle and apoptosis. Eur J Med Chem 2015; 92:302-313.
45. Chan SH, Chui CH, Chan SW, Kok SHL, Chan D, Tsoi MYT, et al. Synthesis of 8-hydroxyquinoline derivatives as novel antitumor agents. ACS Med Chemlett 2012; 4:170-174.