Sains Malaysiana 53(11)(2024): 3673-3682
http://doi.org/10.17576/jsm-2024-5311-11
Unveiling the Cytotoxic and
Apoptosis-Inducing Abilities of an Edible Bird’s Nest Extract in Human
Hepatocellular Carcinoma Huh-7 Cells
(Mendedahkan Keupayaan Aruhan Sitotoksik dan Apoptosis Ekstrak Sarang Burung dalam Sel Karsinoma Hepatosel Manusia Huh-7)
THI-PHUONG NGUYEN1,§,
TANG VAN DUONG2,§,
HOANG DANG KHOA DO1, VAN MINH LE3, KIM NHUNG TA4 & KHOA THI NGUYEN1,*
1NTT Hi-Tech Institute, Nguyen Tat Thanh University, Ho Chi
Minh City 70000, Vietnam
2Vietnam National Museum of Nature, Vietnam Academy of
Science and Technology, 18 Hoang Quoc Viet, Nghia Do, Cau Giay, Vietnam
3Research Center of Ginseng and Medicinal Materials
(CGMM), National Institute of Medicinal Materials, Ho Chi Minh City 700000,
Vietnam
4VNU Vietnam Japan University,
Hanoi, Hanoi 100000, Vietnam
Received: 7
November 2023/Accepted: 19 September 2024
§These authors contributed
equally to the work
Abstract
Edible bird’s nest (EBN) is renowned
for diverse pharmacological properties; however, its ability to cause cancer cell death has
not been found in recent studies. This study investigates the phytochemical composition of EBN ethanol extract (EEE), the
cytotoxic and apoptosis-inducing effects of the extract on human hepatocellular
carcinoma Huh-7 cells. Qualitative and quantitative
tests showed that proteins and peptides were abundant in the EEE. In the MTT
assay, the extract exhibited cytotoxic activity against Huh-7 cells (IC50 = 58.57 ± 2.66 µg/mL, 53.15 ± 3.07 µg/mL, and 55.27 ±
1.85 µg/mL at 24 h, 48 h, and 72 h of incubation, respectively). Annexin-V/PI staining suggested that the extract
provoked apoptosis in Huh-7 cells by deregulating the expression patterns of
apoptotic markers including caspase 3, caspase
8, caspase 9, and survivin. These findings
demonstrate the potential of the EEE as a promising anticancer candidate for
further studies.
Keywords: Apoptosis; cytotoxicity; edible bird’s nest; human hepatocellular carcinoma
Abstrak
Sarang burung (EBN) terkenal dengan
sifat farmakologi yang pelbagai; walau bagaimanapun, keupayaannya untuk
menyebabkan kematian sel kanser masih tidak diketahui. Penyelidikan ini
mengkaji komposisi fitokimia ekstrak etanol EBN (EEE) dan kesan aruhan ekstrak
terhadap aktiviti sitotoksik dan apoptosis pada sel karsinoma hepatosel manusia
Huh-7. Ujian kualitatif dan kuantitatif menunjukkan bahawa protein dan peptida
banyak terdapat dalam EEE. Dalam ujian MTT, ekstrak menunjukkan aktiviti sitotoksik terhadap sel Huh-7 (IC50 = 58.57 ± 2.66 µg/mL, 53.15 ± 3.07 µg/mL dan 55.27 ± 1.85 µg/mL masing-masing
pada 24 jam, 48 jam dan 28 jam eraman). Pewarnaan Annexin-V / PI mencadangkan
bahawa ekstrak EEE mencetuskan apoptosis dalam sel Huh-7 dengan menyahkawal
atur corak pengekspresan penanda apoptosis termasuk caspase 3, caspase 8,
caspase 9 dan survivin. Penemuan ini menunjukkan potensi EEE sebagai calon
antikanser yang berpotensi untuk kajian lanjut.
Kata kunci: Apoptosis; karsinoma hepatosel manusia; kesitotoksikan; sarang burung
REFERENCES
Anthis, N.J. & Clore, G.M. 2013.
Sequence‐specific determination of protein and peptide concentrations by
absorbance at 205 nm. Protein Science 22(6): 851-858.
Auwal, M.S., Saka, S., Mairiga, I.A.,
Sanda, K.A., Shuaibu, A. & Ibrahim, A. 2014. Preliminary phytochemical and
elemental analysis of aqueous and fractionated pod extracts of Acacia nilotica
(Thorn mimosa). Veterinary
Research Forum 5(2): 95-100.
Bradford, M.M. 1976. A rapid and
sensitive method for the quantitation of microgram quantities of protein
utilizing the principle of protein-dye binding. Analytical Biochemistry 72(1-2): 248-254.
Braekman, J.C., Daloze, D. &
Pasteels, J. 1998. Alkaloids in animals. In Alkaloids:
Biochemistry, Ecology, and Medicinal Applications, edited by Roberts, M.F.
& Wink, M. New York: Springer. pp. 349-378.
Ciftci, H.I., Can, M., Ellakwa, D.E.,
Suner, S.C., Ibrahim, M.A., Oral, A., Sekeroglu, N., Özalp, B., Otsuka, M.
& Fujita, M. 2020. Anticancer activity of Turkish marine extracts: A purple
sponge extract induces apoptosis with multitarget kinase inhibition activity. Investigational New Drugs 38: 1326-1333.
Das, B., Al-Amin, M., Russel, S.,
Kabir, S., Bhattacherjee, R. & Hannan, J. 2014. Phytochemical screening and
evaluation of analgesic activity of Oroxylum indicum. Indian Journal of Pharmaceutical Sciences 76(6): 571-575.
Fan, Q., Lian, J., Liu, X., Zou, F.,
Wang, X. & Chen, M. 2021. A study on the skin whitening activity of Digesta
from edible bird’s nest: A mucin glycoprotein. Gels 8(1): 24. https://doi.org/https://doi.org/10.3390/gels8010024
Gheraibia, S., Belattar, N. &
Abdel-Wahhab, M.A. 2020. HPLC analysis, antioxidant and cytotoxic activity of
different extracts of Costus speciosus against HepG-2 cell lines. South African Journal of Botany 131:
222-228.
Hsu, K.C., Li-Chan, E.C. & Jao,
C.L. 2011. Antiproliferative activity of peptides prepared from enzymatic
hydrolysates of tuna dark muscle on human breast cancer cell line MCF-7. Food Chemistry 126(2): 617-622.
Jiang, S., Liu, S., Zhao, C. &
Wu, C. 2016. Developing protocols of tricine-SDS-PAGE for separation of
polypeptides in the mass range 1-30 kDa with minigel electrophoresis system. International Journal of Electrochemical
Science 11: 640-649.
Kancherla, N., Dhakshinamoothi, A.,
Chitra, K. & Komaram, R.B. 2019. Preliminary analysis of phytoconstituents
and evaluation of anthelminthic property of Cayratia auriculata (in
vitro). Maedica 14(4): 350-356.
Khanavi, M., Nabavi, M., Sadati, N.,
Shams Ardekani, M., Sohrabipour, J., Nabavi, S.M.B., Ghaeli, P. & Ostad,
S.N. 2010. Cytotoxic activity of some marine brown algae against cancer cell
lines. Biological Research 43(1):
31-37.
Krelle, C.A., Okoli, S.A. &
Mendz, L.G. 2013. Huh-7 human liver cancer cells: A model system to understand
hepatocellular carcinoma and therapy. Journal
of Cancer Therapy 4(2): 606-631.
Kumar, P., Nagarajan, A. & Uchil,
P. 2018. Analysis of cell viability by the MTT assay. Cold Spring Harb Protoc. 2018: prot095505.
https://doi.org/https://doi.org10.1101/pdb.prot095505
Lee, T., Lau, T. & Ng, I. 2002.
Doxorubicin-induced apoptosis and chemosensitivity in hepatoma cell lines. Cancer Chemother. Pharmacol. 49: 78-86.
Lee, T.H., Wani, W.A., Lee, C.H.,
Cheng, K.K., Shreaz, S., Wong, S., Hamdan, N. & Azmi, N.A. 2021. Edible
bird's nest: The functional values of the prized animal-based bioproduct from
Southeast Asia-A review. Frontiers in
Pharmacology 12: 626233.
https://doi.org/https://doi.org/10.3389/fphar.2021.626233
Ma, F. & Liu, D. 2012. Sketch of
the edible bird's nest and its important bioactivities. Food Research International 48(2): 559-567.
Marcone, M.F. 2005. Characterization
of the edible bird’s nest the ‘Caviar of the East’. Food Research International 38(10): 1125-1134.
Meerloo, J.V., Kaspers, G.J. & Cloos, J. 2011. Cell sensitivity assays: The MTT
assay. Methods in Molecular Biology 731: 237-245.
Nguyen, T-P., Phan, H.N., Do, T.D.,
Do, G.D., Ngo, L.H., Do, H.D.K. & Nguyen, K.T. 2023. Polysaccharide and
ethanol extracts of Anoectochilus formosanus Hayata: Antioxidant,
wound-healing, antibacterial, and cytotoxic activities. Heliyon 9(3): e13559.
Nursid, M., Marraskuranto, E. &
Chasanah, E. 2019. Cytotoxicity and apoptosis induction of sea cucumber Holothuria
atra extracts. Pharmacognosy Research 11: 41-46.
https://doi.org/https://doi.org/10.4103/pr.pr_3_18
Pagliara, P., Scarano, A., Barca, A.,
Zuppone, S., Verri, T. & Caroppo, C. 2015. Ostreopsis cf. ovata induces
cytoskeletal disorganization, apoptosis, and gene expression disregulation on
HeLa cells. Journal of Applied Phycology 27: 2321-2332.
Permatasari, H.K., Permatasari, Q.I.,
Taslim, N.A., Subali, D., Kurniawan, R., Surya, R., Qhabibi, F.R., Tanner,
M.J., Batubara, S.C., Mayulu, N., Gunawan, W.B., Syauki, A.Y., Salindeho, N.,
Park, M.N., Lele, J.A.J.M.N., Tjandrawinata, R.R., Kim, B. & Nurkolis, F.
2023. Revealing edible bird nest as novel
functional foods in combating metabolic syndrome: Comprehensive in silico, in vitro, and in vivo studies. Nutrients 15(18): 3886.
Quek, M.C., Chin, N.L., Yusof, Y.A.,
Law, C.L. & Tan, S.W. 2018. Characterization of edible bird's nest of
different production, species and geographical origins using nutritional
composition, physicochemical properties and antioxidant activities. Food Research International 109: 35-43.
Rashed, A.A. & Nazaimoon, W.W.
2010. Effect of edible bird’s nest on Caco-2 cell proliferation. Journal of Food Science and Technology 8(3): 126-130.
Roy, M.K., Kuwabara, Y., Hara, K.,
Watanabe, Y. & Tamai, Y. 2002. Peptides from the N-terminal end of bovine
lactoferrin induce apoptosis in human leukemic (HL-60) cells. Journal of Dairy Science 85(9):
2065-2074.
Schmittgen, T.D. & Livak, K.J.
2008. Analyzing real-time PCR data by the comparative CT method. Nature Protocols 3(6): 1101-1108.
Serna, N., Sánchez-García, L.,
Unzueta, U., Díaz, R., Vázquez, E., Mangues, R. & Villaverde, A. 2018.
Protein-based therapeutic killing for cancer therapies. Trends in Biotechnology 36(3): 318-335.
Shim, E.K., Chandra, G.F., Pedireddy,
S. & Lee, S-Y. 2016. Characterization of swiftlet edible bird nest, a mucin
glycoprotein, and its adulterants by Raman microspectroscopy. Journal of Food Science 53: 3602-3608.
Truong, D-H., Nguyen, D.H., Ta,
N.T.A., Bui, A.V., Do, T.H. & Nguyen, H.C. 2019. Evaluation of the use of
different solvents for phytochemical constituents, antioxidants, and in
vitro anti-inflammatory activities of Severinia buxifolia. Journal of Food Quality 2019: 8178294.
https://doi.org/https://doi.org/10.1155/2019/8178294.
Wang, B., Li, Z-R., Chi, C-F., Zhang,
Q-H. & Luo, H-Y. 2012. Preparation and evaluation of antioxidant peptides
from ethanol-soluble proteins hydrolysate of Sphyrna lewini muscle. Peptides 36(2): 240-250.
Wang, L., Dong, C., Li, X., Han, W.
& Su, X. 2017. Anticancer potential of bioactive peptides from animal
sources. Oncology Reports 38(2):
637-651.
Wang, Y., Zhong, J., Bai, J., Tong,
R., An, F., Jiao, P., He, L., Zeng, D., Long, E. & Yan, J. 2018. The
application of natural products in cancer therapy by targeting apoptosis
pathways. Current Drug Metabolism 19(9): 739-749.
Yew, M.Y., Koh, R.Y., Chye, S.M.,
Othman, I. & Ng, K.Y. 2014. Edible bird’s nest ameliorates oxidative
stress-induced apoptosis in SH-SY5Y human neuroblastoma cells. BMC Complementary Medicine and Therapies 14: 391.
Yida, Z., Imam, M.U. & Ismail, M. 2014. In vitro bioaccessibility and antioxidant properties of edible bird's
nest following simulated human gastro-intestinal digestion. BMC Complementary Medicine and Therapies 14: 468-475.
Zhao, R., Li, G., Kong, X.J., Huang,
X.Y., Li, W., Zeng, Y.Y. & Lai, X.P. 2016. The improvement effects of
edible bird's nest on proliferation and activation of B lymphocyte and its
antagonistic effects on immunosuppression induced by cyclophosphamide. Drug Design, Development and Therapy 10:
371-381.
*Corresponding author; email: khoant@ntt.edu.vn