TY  - JOUR
T1  - Betulinic Acid Induces Apoptosis and G2/M Cell Cycle Arrest in SH-SY5Y Neuroblastoma Cells via EAG1 Suppression
TT  - Betülinik asit EAG1 Baskılanması Yoluyla SH-SY5Y Nöroblastom Hücrelerinde Apoptoz ve G2/M Hücre Döngüsü Durmasını İndükler
AU  - Çarhan, Ahmet
AU  - Yalçınkaya, Tuğba
AU  - Kaya, Şeyda
AU  - Yildiz, Hatice Kubra
PY  - 2025
DA  - August
Y2  - 2025
DO  - 10.29058/mjwbs.1743485
JF  - Medical Journal of Western Black Sea
JO  - Med J West Black Sea
PB  - Zonguldak Bülent Ecevit Üniversitesi
WT  - DergiPark
SN  - 2822-4302
SP  - 274
EP  - 284
VL  - 9
IS  - 2
LA  - en
AB  - Aim: Neuroblastoma is the most common extracranial solid tumour in children and is associated with high mortality, particularly in advancedstages. Given the limitations of current treatment approaches, such as recurrence and drug resistance, novel agents with tumour-specificeffects and minimal toxicity to normal cells are needed. Betulinic Acid (BA), a naturally occurring pentacyclic triterpene, has gained attentiondue to its selective antitumor properties.Material and Methods: This study investigated the cytotoxic and molecular effects of BA on SH-SY5Y neuroblastoma and epithelialcell lines, Vero. Cell viability was assessed using the MTT assay. Microscopic observations were conducted to evaluate morphologicalchanges. Quantitative real-time PCR (qPCR) was performed to analyse the gene expression changes associated with apoptosis (Bax, Bcl-2,Caspase-3, p53, p21) and the cell cycle (CDK1, Cyclin B1, Ki-67, EAG1).Results: BA significantly reduced cell viability in both cell lines in a time- and dose-dependent manner, with SH-SY5Y cells showing greatersensitivity (IC₅₀=75 μM at 48 hours). Morphological changes such as cell shrinkage and reduced cell density were more pronounced inSH-SY5Y cells. In this cell line, BA treatment significantly upregulated pro-apoptotic genes (Bax, Caspase-3, p53, p21) and downregulatedanti-apoptotic and proliferation-related genes (Bcl-2, CDK1, Cyclin B1, Ki-67, EAG1) (p
KW  - Betulinic acid
KW  - neuroblastoma
KW  - SH-SY5Y
KW  - apoptosis
KW  - cell cycle
KW  - EAG1
N2  - Amaç: Nöroblastoma, çocuklarda en sık görülen ekstrakraniyal solid tümör olup özellikle ileri evrelerde yüksek mortaliteye sahiptir.Tekrarlama ve ilaç direnci gibi mevcut tedavi yöntemlerinin sınırlılıkları nedeniyle, tümöre özgü etkisi yüksek ve normal hücrelere minimumtoksisite gösteren yeni ajanlara ihtiyaç vardır. Doğal bir pentasiklik triterpen olan betulinik asit (BA), seçici antitümör özellikleri nedeniyle ilgiçekmektedir.Gereç ve Yöntemler: Bu çalışmada, BA’nın SH-SY5Y nöroblastoma ve epitelyal hücre hattı, Vero, üzerindeki sitotoksik ve moleküler etkileriincelendi. Hücre canlılığı ise MTT testi ile değerlendirildi. Morfolojik değişiklikler mikroskop ile gözlemlendi. Apoptoz (Bax, Bcl-2, Caspase-3,p53, p21) ve hücre döngüsü (CDK1, Cyclin B1, Ki-67, EAG1) ile ilişkili genlerin ekspresyon seviyeleri qPCR ile analiz edildi.Bulgular: BA, her iki hücre hattında da zaman ve doz bağımlı olarak hücre canlılığını anlamlı şekilde azalttı; SH-SY5Y hücreleri dahaduyarlı bulundu (48 saat için IC₅₀=75 μM). SH-SY5Y hücrelerinde hücre büzülmesi ve hücre yoğunluğunda azalma gibi morfolojikdeğişiklikler belirgindi. Bu hücre hattında BA tedavisi pro-apoptotik genlerin (Bax, Caspase-3, p53, p21) ifadesini artırırken, anti-apoptotik veproliferasyonla ilişkili genlerin (Bcl-2, CDK1, Cyclin B1, Ki-67, EAG1) ifadesini azalttı (p
CR  - 1. Turkish Statistical Institute. (2022). Health statistics yearbook
2022 [Data set]. https://data.tuik.gov.tr/Bulten/Index?p=Saglik-
Istatistikleri-Yilligi-2022-46749
CR  - 2. Ministry of Health of Türkiye. (2022). Childhood cancers in Türkiye:
Incidence and evaluation report [PDF]. Republic of Türkiye
Ministry of Health. https://hsgm.saglik.gov.tr/depo/birimler/
kanser-db/Dokumanlar/Raporlar/17.Agustos_2021_Kanser_
Kontrol_Programi_versiyon-1.pdf
CR  - 3. World Health Organization. (2022). Cancer. https://www.who.
int/news-room/fact-sheets/detail/cancer
CR  - 4. Banerjee, S., Banerjee, S., Bishayee, A., Da Silva, M. N., Sukocheva,
O. A., Tse, E., ... & Bishayee, A. (2024). Cellular and
molecular mechanisms underlying the potential of betulinic
acid in cancer prevention and treatment. Phytomedicine, 132,
155858. https://doi.org/10.1016/j.phymed.2024.155858
CR  - 5. Aswathy, M., Vijayan, A., Daimary, U. D., Girisa, S., Radhakrishnan,
K. V., & Kunnumakkara, A. B. (2022). Betulinic acid: A
natural promising anticancer drug, current situation, and future
perspectives. Journal of Biochemical and Molecular Toxicology,
36(12), e23206. https://doi.org/10.1002/jbt.23206
CR  - 6. Gao, S., Wang, W., Ye, W., & Wang, K. (2022). The mechanism
study of Eag1 potassium channel in gastric cancer.
Translational Cancer Research, 11(10), 3827–3835. https://doi.
org/10.21037/tcr-22-2276
CR  - 7. Luis, E., Anaya-Hernández, A., León-Sánchez, P., & Durán-
Pastén, M. L. (2022). The Kv10.1 channel: A promising target
in cancer. International Journal of Molecular Sciences, 23(15),
8458. https://doi.org/10.3390/ijms23158458
CR  - 8. Toplak, Ž., Hendrickx, L. A., Abdelaziz, R., Shi, X., Peigneur, S.,
Tomašič, T., ... & Pardo, L. A. (2022). Overcoming challenges
of HERG potassium channel liability through rational design:
Eag1 inhibitors for cancer treatment. Medicinal Research Reviews,
42(1), 183–226. https://doi.org/10.1002/med.21808
CR  - 9. An, T., Zha, W. & Zi, J. (2020). Biotechnological production of
betulinic acid and derivatives and their applications. Appl Microbiol
Biotechnol 104, 3339–3348. https://doi.org/10.1007/
s00253-020-10495-1
CR  - 10. Hordyjewska, A., Ostapiuk, A., Horecka, A., & Kurzepa, J.
(2019). Betulin and betulinic acid: Triterpenoids derivatives
with a powerful biological potential. Phytochemistry Reviews,
18, 929–951. https://doi.org/10.1007/s11101-019-09623-1
CR  - 11. Bettariga, F., Taaffe, D. R., Galvão, D. A., Bishop, C., Kim,
J. S., & Newton, R. U. (2024). Suppressive effects of exercise-
conditioned serum on cancer cells: A narrative review of
the influence of exercise mode, volume, and intensity. In Journal
of Sport and Health Science (Vol. 13, Issue 4). https://doi.
org/10.1016/j.jshs.2023.12.001
CR  - 12. Zeng, A. Q., Yu, Y., Yao, Y. Q., Yang, F. F., Liao, M., Song, L.
J., Li, Y. L., Yu, Y., Li, Y. J., Deng, Y. Le, Yang, S. P., Zeng, C.
J., Liu, P., Xie, Y. M., Yang, J. L., Zhang, Y. W., Ye, T. H., &
Wei, Y. Q. (2018). Betulinic acid impairs metastasis and reduces
immunosuppressive cells in breast cancer models. Oncotarget,
9(3). https://doi.org/10.18632/oncotarget.23376
CR  - 13. Lombrea, A., Scurtu, A. D., Avram, S., Pavel, I. Z., Turks, M.,
Lugiņina, J., ... & Danciu, C. (2021). Anticancer potential of
betulonic acid derivatives. International Journal of Molecular
Sciences, 22(7), 3676. https://doi.org/10.3390/ijms22073676
CR  - 14. Hordyjewska, A., Ostapiuk, A., & Horecka, A. (2018). Betulin
and betulinic acid in cancer research. Journal of Pre-Clinical
and Clinical Research, 12(2), 72–75. https://bibliotekanauki.pl/
articles/3735.pdf
CR  - 15. Ali Seyed, M., Dhyani, A., & Csuk, R. (2022). Chemopreventive
and chemotherapeutic potential of betulin and betulinic acid:
mechanistic insights from in vitro, in vivo and clinical studies.
Food Science & Nutrition, 10(11), Article 2903. https://doi.
org/10.3390/biomedicines10112903
CR  - 16. Zúñiga, L., Cayo, A., González, W., Vilos, C., & Zúñiga, R.
(2022). Potassium channels as a target for cancer therapy:
Current perspectives. OncoTargets and Therapy, 15, 783–800.
https://doi.org/10.2147/OTT.S346369
UR  - https://doi.org/10.29058/mjwbs.1743485
L1  - https://dergipark.org.tr/tr/download/article-file/5059443
ER  -