Anticancer Effect of Fluorouracil and Gum-Based Cerium Oxide Nanoparticles on Human Malignant Colon Carcinoma Cell Line (Caco2)

Document Type : Original Article

Authors

1 Division of Cell and Molecular Biology, Department of Biology, Faculty of Science, University of Zabol, Zabol, Iran

2 Department of Physics, Faculty of Science, University of Zabol, Zabol, Iran

3 Department of Medical Genetics, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

Abstract

Objective: We investigated whether co-incubation of 5-FU and gum-based cerium oxide nanoparticles (CeO2 NPs) would improve half-maximal inhibitory concentration (IC50) and apoptosis in the Caco-2 cancer cell line
Materials and Methods: In this experimental study, we synthesized Ceo-2-XG by the nano perception method.
X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), transmission electron microscopy
(TEM), dynamic light scattering (DLS) and vibrating sample magnetometer (VSM) techniques were employed
to characterize the synthesized nanoparticles. The Caco-2 cancer cells were cultured and treated with Ceo-2-
XG and 5-FU. Cytotoxicity analysis was carried out using MTT assay on Caco-2 cancer cells. CXCR1, CXCR2,
CXCL8, BAX, BCL-2, P53, CASPASE-3, CASPASE-8 and CASPASE-9 gene expression changes were assessed
by quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR). The Caco-2 cancer cell mortality
mechanism was analyzed using Annexin V-FITC/PI flow cytometry. Using the inverted microscope morphology
changes of the Caco-2 cancer cells was observed.
Results: With a sample size of roughly 11 nm, TEM analysis revealed spherical structures. Interestingly, after 72
hours, 400 μg/ml nanoparticles significantly lowered the IC50 of 5-FU from 101 to 71 μg/ml (P<000.1). Furthermore,
qRT-PCR analysis showed that BCL-2, CXCR1, CXCR2 and CXCR8 expressions were significantly decreased in
the 5-FU and Ceo-2-XG nanoparticles co-incubated group, compared to the 5-FU alone (P<0.001). Notably, gene
expressions of BAX, P53, CASPASE-3, CASPASE-8 and CASPASE-9 were significantly higher in the 5-FU and Ceo-
2-XG nanoparticles co-incubated group, compared to the 5-FU alone (P<0.001). The findings revealed that dead cells
owing to apoptosis were more than two times higher in 5-FU and Ceo-2-XG nanoparticles cancer cells than in 5-FU
alone treated cancer cells.
Conclusion: Co-incubation of 5-FU and Ceo-2-XG nanoparticles significantly increased apoptosis in the Caco-2
cancer cells. The antiproliferative activity of co-incubated 5-FU and Ceo-2-XG nanoparticles on Caco-2 cancer cells
was substantially higher than that of 5-FU alone.

Keywords


  1. Pourmoshir N, Motalleb GH, Vallian S. hsa-miR-423 rs6505162 is associated with the increased risk of breast cancer in isfahan central province of Iran. Cell J. 2020; 22 Suppl 1: 110-116.
  2. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2021; 71(3): 209-249.
  3. Munteanu I, Mastalier B. Genetics of colorectal cancer. J Med Life. 2014; 7(4): 507-511.
  4. Pandey S, De Klerk C, Kim J, Kang M, Fosso-Kankeu E. Eco friendly approach for synthesis, characterization and biological activities of milk protein stabilized silver nanoparticles. Polymers (Basel). 2020; 12(6): 1418.
  5. Li C, Shi X, Shen Q, Guo C, Hou Z, Zhang J. Hot topics and challenges of regenerative nanoceria in application of antioxidant therapy. J Nanomater. 2018; 2018: 1-12.
  6. Elmore S. Apoptosis: a review of programmed cell death. Toxicol Pathol. 2007; 35(4): 495-516.
  7. Singh S, Wu S, Varney M, Singh AP, Singh RK. CXCR1 and CXCR2 silencing modulates CXCL8-dependent endothelial cell proliferation, migration and capillary-like structure formation. Microvasc Res. 2011; 82(3): 318-325.
  8. Rahdar A, Aliahmad M, Hajinezhad MR, Samani M. Xanthan gumstabilized nano-ceria: green chemistry based synthesis, characterization, study of biochemical alterations induced by intraperitonealdoses of nanoparticles in rat. J Mol Struct. 2018; 1173; 166-172.
  9. Levit SL, Yang H, Tang C. Rapid self-assembly of polymer nanoparticles for synergistic codelivery of paclitaxel and lapatinib via flash nanoprecipitation. Nanomaterials (Basel). 2020; 10(3): 561.
  10. Suzuki R, Kang Y, Li X, Roife D, Zhang R, Fleming JB. Genistein potentiates the antitumor effect of 5-Fluorouracil by inducing apoptosis and autophagy in human pancreatic cancer cells. Anticancer Res. 2014; 34(9): 4685-4692.
  11. Sargazi S, Hajinezhad MR, Barani M, Rahdar A, Shahraki S, Karimi P, et al. Synthesis, characterization, toxicity and morphology assessments of newly prepared microemulsion systems for delivery of valproic acid. J Mol Liq. 2021; 338: 116625.
  12. Le Berre M, Gerlach JQ, Dziembała I, Kilcoyne M. Calculating half maximal inhibitory concentration (IC50) values from glycomics microarray data using GraphPad Prism. Methods Mol Biol. 2022; 2460: 89-111.
  13. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-delta delta C(T)) method. Methods. 2001; 25(4): 402-408.
  14. Koressaar T, Remm M. Enhancements and modifications of primer design program Primer3. Bioinformatics. 2007; 23(10): 1289-1291.
  15. Ye J, Coulouris G, Zaretskaya I, Cutcutache I, Rozen S, Madden TL. Primer-BLAST: a tool to design target-specific primers for polymerase chain reaction. BMC Bioinformatics. 2012; 13: 134.
  16. Shen YH, Wang LY, Zhang BB, Hu QM, Wang P, He BQ, et al. Ethyl rosmarinate protects high glucose-induced injury in human endothelial cells. Molecules. 2018; 23(12): 3372.
  17. Heidarzadeh S, Motalleb GH, Zorriehzahra MJ. Evaluation of tumor regulatory genes and apoptotic pathways in the cytotoxic effect of cytochalasin h on malignant human glioma cell line (U87MG). Cell J. 2019; 21(1): 62-69.
  18. Latchman J, Guastella A, Tofthagen C. 5-Fluorouracil toxicity and dihydropyrimidine dehydrogenase enzyme: implications for practice. Clin J Oncol Nurs. 2014; 18(5): 581-585.
  19. Gulbake A, Jain A, Jain A, Jain A, Jain SK. Insight to drug delivery aspects for colorectal cancer. World J Gastroenterol. 2016; 22(2): 582-599.
  20. Brar B, Ranjan K, Palria A, Kumar R, Ghosh M, Sihag S, et al. Nanotechnology in colorectal cancer for precision diagnosis and therapy. Front Nanosci.2021; 3: 699266.
  21. Sisubalan N, Ramkumar VS, Pugazhendhi A, Karthikeyan C, Indira K, Gopinath K, et al. ROS-mediated cytotoxic activity of ZnO and CeO2 nanoparticles synthesized using the Rubia cordifolia L. leaf extract on MG-63 human osteosarcoma cell lines. Environ Sci Pollut Res Int. 2018; 25(11): 10482-10492.
  22. Palaniraj A, Jayaraman V. Production, recovery and applications of xanthan gum by Xanthomonas campestris. J Food Eng. 2011; 106(1): 1-12.
  23. Stone V, Johnston H, Schins RP. Development of in vitro systems for nanotoxicology: methodological considerations. Crit Rev Toxicol. 2009; 39(7): 613-626.
  24. Lee WH, Loo CY, Traini D, Young PM. Inhalation of nanoparticlesbased drug for lung cancer treatment: advantages and challenges. Asian J Pharm Sci. 2015; 10: 481-489.
  25. Su Y, Hu J, Huang Z, Huang Y, Peng B, Xie N, et al. Paclitaxelloaded star-shaped copolymer nanoparticles for enhanced malignant melanoma chemotherapy against multidrug resistance. Drug Des Devel Ther. 2017; 11: 659-668.
  26. Danial NN, Korsmeyer SJ. Cell death: critical control points. Cell. 2004; 116(2): 205-219.
  27. Bauer JH, Helfand SL. New tricks of an old molecule: lifespan regulation by p53. Aging Cell. 2006; 5(5): 437-440.
  28. Naseri MH, Mahdavi M, Davoodi J, Tackallou SH, Goudarzvand M, Neishabouri SH. Up regulation of Bax and down regulation of Bcl2 during 3-NC mediated apoptosis in human cancer cells. Cancer Cell Int. 2015; 15: 55.
  29. Aali N, Motalleb G. The effect of nicotine on the expressions of the α7 nicotinic receptor gene and Bax and Bcl-2 proteins in the mammary gland epithelial-7 breast cancer cell line and its relationship to drug resistance. Cell Mol Biol Lett. 2015; 20(5): 948-964.
  30. Labi V, Erlacher M. How cell death shapes cancer. Cell Death Dis. 2015; 6(3): e1675.
  31. McIlwain DR, Berger T, Mak TW. Caspase functions in cell death and disease. Cold Spring Harb Perspect Biol. 2013; 5(4): a008656.
  32. Ha H, Debnath B, Neamati N. Role of the CXCL8-CXCR1/2 axis in cancer and inflammatory diseases. Theranostics. 2017; 7(6): 1543-1588.
  33. Erreni M, Bianchi P, Laghi L, Mirolo M, Fabbri M, Locati M, et al. Expression of chemokines and chemokine receptors in human colon cancer. Methods Enzymol. 2009; 460: 105-121.
  34. Wen Y, Giardina SF, Hamming D, Greenman J, Zachariah E, Bacolod MD, et al. GROalpha is highly expressed in adenocarcinoma of the colon and down-regulates fibulin-1. Clin Cancer Res. 2006; 12(20 Pt 1): 5951-5959.
  35. Ogata H, Sekikawa A, Yamagishi H, Ichikawa K, Tomita S, Imura J, et al. GROα promotes invasion of colorectal cancer cells. Oncol Rep. 2010; 24(6): 1479-1486.
  36. Sharma B, Nawandar DM, Nannuru KC, Varney ML, Singh RK. Targeting CXCR2 enhances chemotherapeutic response, inhibits mammary tumor growth, angiogenesis, and lung metastasis. Mol Cancer Ther. 2013; 12(5): 799-808.
  37. Nannuru KC, Sharma B, Varney ML, Singh RK. Role of chemokine receptor CXCR2 expression in mammary tumor growth, angiogenesis and metastasis. J Carcinog. 2011; 10: 40.
  38. Ning Y, Manegold PC, Hong YK, Zhang W, Pohl A, Lurje G, et al. Interleukin-8 is associated with proliferation, migration, angiogenesis and chemosensitivity in vitro and in vivo in colon cancer cell line models. Int J Cancer. 2011; 128(9): 2038-2049.