The Effect of ZnO Nanoparticles Functionalized with Glutamic Acid and Conjugated with Thiosemicarbazide on the Expression of Bax, caspase-3, and bcl-2 Genes in Adenocarcinoma Gastric (AGS) Cell Line

Subject Areas : Journal of Animal Biology

Sadaf Beigi ¹ , Ali Salehzadeh ² , Hadi Habibbollahi ³ , Seyed Ataollah Sadat Shandiz ⁴ , Fariba Safa ⁵

1 - Department of Biology, Rasht Branch, Islamic Azad University, Rasht, Iran
2 - Department of Biology, Rasht Branch, Islamic Azad University, Rasht, Iran
3 - Department of Biology, Rasht Branch, Islamic Azad University, Rasht, Iran
4 - Department of Biology, Central Tehran Branch, Islamic Azad University, Tehran, Iran
5 - Department of Chemistry, Rasht Branch, Islamic Azad University, Rasht, Iran

Received: 2021-07-28 Accepted : 2021-11-24 Published : 2022-08-23

Keywords: gastric cancer, Thiosemicarbazide, anticancer, Zinc Oxide Nanoparticles,

Abstract :

Conjugation of metal nanoparticles (NPs) to thiosemicarbazide improves their stability and anticancer potential. This work was aimed at preparing ZnO NPs functionalized with glutamic acid conjugated to thiosemicarbazide (Zn@Glu/TSC). Then, the anticancer potential of nanoparticles was studied by evaluating their cytotoxicity and the expression of the genes involved in cell apoptosis in AGS cancer cells. Physicochemical properties of the NPs were characterized using FT-IR, SEM, and TEM imaging, and Zeta potential assays. Cytotoxicity and inhibitory concentration 50% value of the NPs was determined using the MTT assay. Moreover, the expression of the caspase-3, bax, and bcl-2 genes among NPs treated AGS cells was investigated using Real-time PCR assay. According to the results, the NPs were spherical with a size range of 10-90 nm. FT-IR assay confirmed the proper synthesis of the NPs and the zeta potential was determined -11.7 mV. The synthesized NPs showed significant cytotoxicity at concentrations ≥ 15.62 µg/mL and 50% inhibitory concentration was determined 90µg/mL. Besides, the expression of caspase-3 and bax genes in NPs treated cells increased by 1.67 and 1.80 folds, respectively, while the bcl-2 gene was regulated by 0.70 folds, compared to the control cells. 

References:

• Akhtar M. J., Ahamed M., Kumar S., Khan M. M., Ahmad J., Alrokayan, S. A. 2012. Zinc oxide nanoparticles selectively induce apoptosis in human cancer cells through reactive oxygen species. International Journal of Nanomedicine, 7: 845.


• Barrak H., Saied T., Chevallier P., Laroche G., M’nif A., Hamzaoui A. H. 2019. Synthesis, characterization, and functionalization of ZnO nanoparticles by N-(trimethoxysilylpropyl) ethylenediamine triacetic acid (TMSEDTA): Investigation of the interactions between Phloroglucinol and ZnO@ TMSEDTA. Arabian Journal of Chemistry, 12(8): 4340-4347.


• Bejarbaneh M., Moradi-Shoeili Z., Jalali, A. Salehzadeh A. 2020. Synthesis of Cobalt Hydroxide Nano-flakes Functionalized with Glutamic Acid and Conjugated with Thiosemicarbazide for Anticancer Activities Against Human Breast Cancer Cells. Biological Trace Element Research, 198(1): 98-108.


• Bendale Y., Bendale V., Paul S. 2017. Evaluation of cytotoxic activity of platinum nanoparticles against normal and cancer cells and its anticancer potential through induction of apoptosis. Integrative medicine research, 6(2): 141-148.


• Bigdeli R., Shahnazari M., Panahnejad E., Cohan R. A., Dashbolaghi A., Asgary V. 2019. Cytotoxic and apoptotic properties of silver chloride nanoparticles synthesized using Escherichia coli cell-free supernatant on human breast cancer MCF 7 cell line. Artificial Cells, Nanomedicine, and Biotechnology, 47(1): 1603-1609.


• Carcas L. P. 2014. Gastric cancer review. Journal of Carcinogenesis, 13: 14.


• Chen X., Yang Q., Chen J., Zhang P., Huang, Q., Zhang, X., Yang L., Xu D., Zhao C., Wang X., Liu, J. 2018. Inhibition of proteasomal deubiquitinase by silver complex induces apoptosis in non-small cell lung cancer cells. Cellular Physiology and Biochemistry, 49(2): 780-797.


• Farmanfarma, K.K, Mahdavifar, N., Hassanipour S., Salehiniya H. 2020. Epidemiologic Study of Gastric Cancer in Iran: A Systematic Review. Clinical and experimental gastroenterology, 13: 511–542.


• Habibzadeh S.Z., Salehzadeh A., Moradi-Shoeili Z., Shandiz S.A.S. 2020. A novel bioactive nanoparticle synthesized by conjugation of 3-chloropropyl trimethoxy silane functionalized Fe 3 O 4 and 1-((3-(4-chlorophenyl)-1-phenyl-1H-pyrazol-4-yl) methylene)-2-(4-phenylthiazol-2-yl) hydrazine: assessment on anti-cancer against gastric AGS cancer cells. Molecular Biology Reports, 47(3): 1637-1647.


• Hamrayev H., Shameli K., Yusefi M. 2020. Preparation of zinc oxide nanoparticles and its cancer treatment effects: A review paper. Journal of Advanced Research in Micro and Nano Engineering, 2(1): 1-11.


• Jalili A., Irani S., Mirfakhraie R. 2016. Combination of cold atmospheric plasma and iron nanoparticles in breast cancer: Gene expression and apoptosis study. OncoTargets and therapy, 9: 5911.


• Jamdagni P., Khatri P., Rana J. S. 2018. Green synthesis of zinc oxide nanoparticles using flower extract of Nyctanthes arbor-tristis and their antifungal activity. Journal of King Saud University-Science, 30(2): 168-175.


• Karuppaiya P., Satheeshkumar E., Tsay H. S. 2019. Biogenic synthesis of silver nanoparticles using rhizome extract of Dysosma pleiantha and its antiproliferative effect against breast and human gastric cancer cells. Molecular biology reports, 46(5): 4725-4734.


• Liao W., Lu Y., Fu J., Ning Z., Yang J., Ren J. 2015. Preparation and characterization of dictyophora indusiata polysaccharide–zinc complex and its augmented antiproliferative activity on human cancer cells. Journal of agricultural and food chemistry, 63(29): 6525-6534.


• Lodish H.F. 2000. Molecular cell biology(4th ed.) New York: W.H. Freeman.


• Ma Z. Y., Shao J., Bao W. G., Qiang Z. Y., Xu J. Y. 2015. A thiosemicarbazone copper (II) complex as a potential anticancer agent. Journal of Coordination Chemistry, 68(2): 277-294.


• Nutting C.M., Van Herpen C.M.L., Miah A.B., Bhide S.A., Machiels J.P., Buter , Kelly C., De Raucourt D., Harrington K.J. 2009. Phase II study of 3-AP Triapine in patients with recurrent or metastatic head and neck squamous cell carcinoma. Annals of oncology, 20(7): 1275-1279.


• Oliveira D.M., Lacava Z.G., Lima E.C., Morais P.C., Tedesco A.C. 2006. Zinc phthalocyanine/magnetic fluid complex: a promising dual nanostructured system for cancer treatment. Journal of nanoscience and nanotechnology, 6(8): 2432-2437.


• Pawlukojć A., Hołderna-Natkaniec K., Bator G., Natkaniec I. 2014. L-glutamine: Dynamical properties investigation by means of INS, IR, RAMAN, 1H NMR and DFT techniques. Chemical physics, 443: 17-25.


• Pelosi G. 2010. Thiosemicarbazone metal complexes: from structure to activity. The Open Crystallography Journal, 3(1).


• Rameshthangam P., Chitra J. P. 2018. Synergistic anticancer effect of green synthesized nickel nanoparticles and quercetin extracted from Ocimum sanctum leaf extract. Journal of materials science & technology, 34(3): 508-522.


• Richardson D. R., Sharpe P. C., Lovejoy D. B., Senaratne D., Kalinowski D. S., Islam M., Bernhardt P. V. 2006. Dipyridyl thiosemicarbazone chelators with potent and selective antitumor activity form iron complexes with redox activity. Journal of medicinal chemistry, 49(22): 6510-6521.


• Senthilkumar S. R., Sivakumar T. 2014. Green tea (Camellia sinensis) mediated synthesis of zinc oxide (ZnO) nanoparticles and studies on their antimicrobial activities. International Journal of Pharmacy and Pharmaceutical Sciences,6(6): 461-465.


• Shandiz S. A. S., Montazeri A., Abdolhosseini M., Shahrestani S. H., Hedayati M., Moradi-Shoeili Z., Salehzadeh A. 2018. Functionalization of Ag nanoparticles by glutamic acid and conjugation of Ag@ Glu by thiosemicarbazide enhances the apoptosis of human breast cancer MCF-7 cells. Journal of Cluster Science, 29(6): 1107-1114.


• Sinha S. K., Ram S., Lamba O. P. 1988. Vibrational (ir and Raman) spectra and conformational studies of 1-formyl-3- thiosemicarbazide. Spectrochimica Acta Part A: Molecular Spectroscopy, 44(7): 713-721.


• Xu Y., Liu L., Qiu X., Liu Z., Li H., Li Z., Luo W., Wang E. 2012. CCL21/CCR7 prevents apoptosis via the ERK pathway in human non-small cell lung cancer cells. PloS one, 7(3):