Evaluation of Cytotoxicity of Gold Glyconanoparticles of Human Colon Adenocarcinoma Cells

Abstract

INTRODUCTION: Use of metal nanoparticles is a promising trend in oncotherapy.

AIM: To evaluate cytotoxicity of gold nanoparticles (GNPs) modified with fucose, lactose and galactose residues, on Caco-2 cells.

MATERIALS AND METHODS: Cells were cultured to a monolayer with GHPs in concentrations of 10 µg/ml–600 µg/ml for particles with fucose, 10 µg/ml–900 µg/ml for particles with lactose and 10 µg/ml–550 µg/ml for particles with galactose. As non-tumor cells, Caco-2 cells were used after differentiation into small-intestine enterocytes (incubation time — 21 days). The cytotoxicity of GNPs was evaluated in MTT-test after incubation with cells for 2 hours, 8 hours and 24 hours. A comparison drug was fluorouracil in concentrations of 50 µg/ml–2000 µg/ml. The half-maximal inhibitory concentration (IC₅₀) was calculated in GraphPad Prism 8.4.3 program.

RESULTS: IC₅₀ for GNPs with fucose for tumor cells was 582 ± 29 and 336 ± 36 µg/ml after incubation for 2 hours and 8 hours, respectively; for GNPs with lactose — 769 ± 50 and 515 ± 45 µg/ml, for GNPs with galactose — 467 ± 299 and 299 ± 28 µg/ml. For differentiated cells, IC₅₀ for all types of GNPs was higher than for tumor cells after incubation for 2 and 8 hours: for particles with fucose — 530 ± 3 and 410 ± 15 µg/ml; for particles with lactose — 831 ± 7 and 639 ± 14 µg/ml; for particles with galactose — 511 ± 21 (p = 0.018) and 376 ± 36 µg/ml, respectively. After 24-hour incubation of Caco-2 cells with fucose, IC50 did not differ for tumor and differentiated varieties. For other GNPs, IC₅₀ for tumor cells was higher than for differentiated cells. Fluorouracil showed cytotoxicity for tumor cells with IC₅₀ 2108 ± 19 and 1764 ± 193 µg/ml after 2 hours and 8 hours of incubation, respectively. For differentiated cells, IC₅₀ of fluorouracil was 1694 ± 102 µg/ml after 2-hour incubation and 1694 ± 102 µg/ml after 8-hour incubation. There were no differences between IC₅₀ of fluorouracil for differentiated and tumor cells with equal incubation time.

CONCLUSION: GNPs with fucose, lactose and galactose have a cytotoxic effect on human colon adenocarcinoma cells and in the activity are superior to fluorouracil.

Keywords

gold glyconanoparticles / human colon adenocarcinoma cells / Caco-2 / fluorourcil / МТТ-test

References

[1] Boytsov SA, Deev AD, Shalnova SA. Mortality and risk factors for non-communicable diseases in Russia: Specific features, trends, and prognosis. Terapevticheskii Arkhiv. 2017;89(1):5–13. (In Russ). doi: 10.17116/terarkh20178915-13

[2] Бойцов С.А., Деев А.Д., Шальнова С.А. Смертность и факторы риска неинфекционных заболеваний в России: особенности, динамика, прогноз // Терапевтический архив. 2017. Т. 89, № 1. С. 5–13. doi: 10.17116/terarkh20178915-13

[3] The 5th Anniversary International Forum of Oncology and Radiotherapy. Moscow, 19–23 September 2022. Available at: https://drive.google.com/file/d/1-wylUcOIASC8Rnsc1BAAPit8kuy9MEGJ/view. Accessed: 2022 November 11. (In Russ).

[4] V Юбилейный международный форум онкологии и радиотерапии. Москва, 23 сентября 2022 г. Доступно по: https://drive.google.com/file/d/1-wylUcOIASC8Rnsc1BAAPit8kuy9MEGJ/view. Ссылка активна на 11.11.2022.

[5] Raguzin EV, Yudin MA, Glushenko DD, et al. Analysis and evaluation of modern approaches to development of medical drugs using micro- and nanotechnologies. I. P. Pavlov Russian Medical Biological Herald. 2022;30(3):397–410. (In Russ). doi: 10.17816/PAVLOVJ104787

[6] Рагузин Е.В., Юдин М.А., Глушенко Н.Г., и др. Анализ и оценка современных подходов к созданию лекарственных средств с использованием микро- и нанотехнологий // Российский медико-биологический вестник имени академика И. П. Павлова. 2022. Т. 3, № 3. С. 397–410. doi: 10.17816/PAVLOVJ104787

[7] Ershov AYu, Kopanitsa MA, Korotkova NV, et al. Antioxidant activity of silver glyconanoparticles based on mercaptopropionylhydrazones of mono- and disaccharides. Nauka Molodykh (Eruditio Juvenium). 2019;7(2):247–54. (In Russ). doi: 10.23888/HMJ201972247-254

[8] Ершов А.Ю., Копаница М.А., Короткова Н.В., и др. Антиоксидантная активность гликонаночастиц серебра на основе меркаптопропионилгидразонов моно- и дисахаридов // Наука Молодых (Eruditio Juvenium). 2019. Т. 7, № 2. С. 247–254. doi: 10.23888/HMJ201972247-254

[9] Kurapov PB, Bakhtenko EYu. Gold nanoparticles in the diagnosis and treatment of cancer. Bulletin of RSMU. 2018;(6):86−93. (In Russ). doi: 10.24075/vrgmu.2018.090

[10] Курапов П.Б., Бахтенко Е.Ю. Наночастицы золота для диагностики и терапии онкологических заболеваний // Вестник РГМУ. 2018. № 6. С. 86−93. doi: 10.24075/vrgmu.2018.090

[11] Ershov AYu, Martynenkov AA, Lagoda IV, et al. Synthesis of aldose 11-mercaptoundecanoyl hydrazones as promising glycoligands of noble metal nanoparticles. ChemistrySelect. 2019;4(44):12938–41. doi: 10.1002/slct.201902942

[12] Ershov A.Yu., Martynenkov A.A., Lagoda I.V., et al. Synthesis of aldose 11-mercaptoundecanoyl hydrazones as promising glycoligands of noble metal nanoparticles // ChemistrySelect. 2019. Vol. 4, No. 44. P. 12938–12941. doi: 10.1002/slct.201902942

[13] Ershov AYu, Martynenkov AA, Lagoda IV, et al. Synthesis of the Gold Glyconanoparticles Based on 6-Deoxyand 2-(Acetylamino)aldoses 3-Mercaptopropionylhydrazones. Russian Journal of General Chemistry. 2021;91(2):260−8. (In Russ). doi: 10.31857/S0044460X21020098

[14] Ершов А.Ю., Мартыненков А.А., Лагода И.В., и др. Синтез гликонаночастиц золота на основе 3-меркаптопропионилгидразонов 6-дезокси- и 2-(ацетиламино)альдоз // Журнал общей химии. 2021. Т. 91, № 2. С. 260−268. doi: 10.31857/S0044460X21020098

[15] Blondy S, David V, Verdier M, et al. 5-Fluorouracil resistance mechanisms in colorectal cancer: From classical pathways to promising processes. Cancer Sci. 2020;111(9):3142–54. doi: 10.1111/cas.14532

[16] Blondy S., David V., Verdier M., et al. 5-Fluorouracil resistance mechanisms in colorectal cancer: From classical pathways to promising processes // Cancer Science. 2020. Vol. 111, No. 9. P. 3142–3154. doi: 10.1111/cas.14532

[17] Sambuy Y, De Angelis I, Ranaldi G, et al. The Caco-2 cell line as a model of the intestinal barrier: influence of cell and culture-related factors on Caco-2 cell functional characteristics. Cell Biol Toxicol. 2005;21(1):1−26. doi: 10.1007/s10565-005-0085-6

[18] Sambuy Y., De Angelis I., Ranaldi G., et al. The Caco-2 cell line as a model of the intestinal barrier: influence of cell and culture-related factors on Caco-2 cell functional characteristics // Cell Biology and Toxicology. 2005. Vol. 21, No. 1. P. 1−26. doi: 10.1007/s10565-005-0085-6

[19] Catanzaro E, Calcabrini C, Bishayee A, et al. Antitumor potential of marine and freshwater lectins. Mar Drugs. 2019;18(1):11. doi: 10.3390/md18010011

[20] Catanzaro E., Calcabrini C., Bishayee A., et al. Antitumor Potential of Marine and Freshwater Lectins // Marine Drugs. 2019. Vol. 18, No. 1. P. 11. doi: 10.3390/md18010011

[21] Setyawati MI, Tay CY, Bay BH, et al. Gold nanoparticles induced endothelial leakiness depends on particle size and endothelial cell origin. ACS Nano. 2017;11(5):5020−30. doi: 10.1021/acsnano.7b01744

[22] Setyawati M.I., Tay C.Y., Bay B.H., et al. Gold nanoparticles induced endothelial leakiness depends on particle size and endothelial cell origin // ACS Nano. 2017. Vol. 11, No. 5. P. 5020−5030. doi: 10.1021/acsnano.7b01744

[23] Pagliaccia MC, Spinozzia F, Miglioratib G, et al. Genistein inhibits tumour cell growth in vitro but enhances mitochondrial reduction of tetrazolium salts: A further pitfall in the use of the MTT assay for evaluating cell growth and survival. Eur J Cancer. 1993;29A(11):1573–7. doi: 10.1016/0959-8049(93)90297-s

[24] Pagliaccia M.C., Spinozzia F., Miglioratib G., et al. Genistein inhibits tumour cell growth in vitro but enhances mitochondrial reduction of tetrazolium salts: A further pitfall in the use of the MTT assay for evaluating cell growth and survival // European Journal of Cancer. 1993. Vol. 29A, No. 11. P. 1573–1577. doi: 10.1016/0959-8049(93)90297-s

[25] Vellonen K–S, Honkakoski P, Urtti A. Substrates and inhibitors of efflux proteins interfere with the MTT assay in cells and may lead to underestimation of drug toxicity. Eur J Pharm Sci. 2004;23(2):181–8. doi: 10.1016/j.ejps.2004.07.006

[26] Vellonen K.–S., Honkakoski P., Urtti A. Substrates and inhibitors of efflux proteins interfere with the MTT assay in cells and may lead to underestimation of drug toxicity // European Journal of Pharmaceutical Sciences. 2004. Vol. 23, No. 2. P. 181–188. doi: 10.1016/j.ejps.2004.07.006

[27] Osuga T, Takimoto R, Ono M, et al. Relationship between increased fucosylation and metastatic potential in colorectal cancer. J Natl Cancer Inst. 2016;108(8):djw21. doi: 10.1093/jnci/djw21

[28] Osuga T., Takimoto R., Ono M., et al. Relationship Between Increased Fucosylation and Metastatic Potential in Colorectal Cancer // Journal of the National Cancer Institute. 2016. Vol. 108, No. 8. P. djw038. doi: 10.1093/jnci/djw21

[29] Yoshida M, Takimoto R, Murase K, et al. Targeting anticancer drug delivery to pancreatic cancer cells using a fucose-bound nanoparticle approach. PLoS One. 2012;7(7):e39545. doi: 10.1371/journal.pone.0039545

[30] Yoshida M., Takimoto R., Murase K., et al. Targeting anticancer drug delivery to pancreatic cancer cells using a fucose-bound nanoparticle approach // PLoS One. 2012. Vol. 7, No. 7. P. e39545. doi: 10.1371/journal.pone.0039545

[31] Kavunja HW, Voss PG, Wang JL, et al. Identification of lectins from metastatic cancer cells through magnetic glyconanoparticles. Isr J Chem. 2015;55(3−4):423–36. doi: 10.1002/IJCH.201400156

[32] Kavunja H.W., Voss P.G., Wang J.L., et al. Identification of lectins from metastatic cancer cells through magnetic glyconanoparticles // Israel Journal of Chemistry. 2015. Vol. 55, No. 3−4. P. 423–436. doi: 10.1002/IJCH.201400156

[33] Casale J, Patel P. Fluorouracil. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2022. Available at: https://www.ncbi.nlm.nih.gov/books/NBK549808/. Accessed: 2022 November 11.

[34] Casale J., Patel P. Fluorouracil. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2023. Доступно по: https://www.ncbi.nlm.nih.gov/books/NBK549808/. Ссылка активна на 11.11.2022.

[35] Prince GT, Cameron MC, Fathi R, et al. Topical 5-fluorouracil in dermatologic disease. Int J Dermatol. 2018;57(10):1259–64. doi: 10.1111/ijd.14106

[36] Prince G.T., Cameron M.C., Fathi R., et al. Topical 5-fluorouracil in dermatologic disease // International Journal of Dermatology. 2018. Vol. 57, No. 10. P. 1259–1264. doi: 10.1111/ijd.14106

RIGHTS & PERMISSIONS

Chernykh I.V., Kopanitsa M.A., Shchul'kin A.V., Yakusheva E.N., Ershov A.Y., Martynenkov A.A., Lagoda I.V., Volkova A.M.