Galium aparine L. extract inhibits the proliferation of HT-29 cells and induces apoptosis

Ayşegül Öztürk; Arzuhan Çetindağ; Fatih Yulak; Sebahattin Karabulut; Ahmet Kemal Filiz

doi:10.66235/kumj.1906992

Galium aparine L. extract inhibits the proliferation of HT-29 cells and induces apoptosis

Abstract

Aims: Galium Aparine L. (G. aparine) has been traditionally used as a medicinal herb for the treatment of cancer, fever, hypertension. This study aimed to evaluate the potential anticancer effects of G. aparine in HT-29 colon cancer cells and to investigate the molecular mechanisms that may be involved in these effects.

Methods: The LC/Q-TOF/MS analysis was employed to conduct a qualitative phytochemical characterization of G. aparine. To evaluate the potential anticolorectal cancer effects of G. aparine, HT-29 cells were pretreated with G. aparine. Then, cell viability was examined by XTT assay, cell cycle distribution by flow cytometry and cell death by annexin V assay. In addition, the effect of G. aparine treatment on epidermal growth factor receptor (EGFR), extracellular signal-regulated kinase- 1 (ERK-1), ERK-2, and apoptotic markers caspase-3 and Bcl-2-associated agonist of cell death (BAD) levels were investigated by ELISA method.

Results: Following treatment with G. aparine, HT-29 cells exhibit increased apoptosis and cell cycle inhibition at the G0/G1 phase, as evidenced by the Annexin V binding assay (p<0.05). Moreever, G. aparine treatment significantly decreased EGFR and ERK-1 levels (p<0.05), while the reduction in ERK-2 level was not statistically significant (p>0.05). The apoptotic impact of G. aparine was additionally verified through ELISA assays evaluating the levels of BAD and caspase 3 (p<0.05).

Conclusion: Taken together, the results demonstrate that G. aparine extract exerts significant anti-colorectal cancer effects by promoting apoptosis and inhibiting EGFR/ERK signaling.

Keywords

References

Ataseven D, Öztürk A, Özkaraca M, Joha Z. Anticancer activity of lycopene in HT-29 colon cancer cell line. Medical Oncology. 2023;40(5):127. doi:10.1007/s12032-023-02001-0
Jourand KJ, Davies J. Colorectal cancer: features and investigation. Medicine. 2024;52(5):258-262. doi:10.1016/J.-MPMED.2024.02.008
Orhan N, Deliorman Orhan D, Aslan M, Ükürolu M, Orhan IE. UPLC-TOF-MS analysis of Galium spurium towards its neuroprotective and anticonvulsant activities. J Ethnopharmacol. 2012;141(1):220-227. doi:10.1016/j.jep.2012.01.056
Sahin B, Karabulut S, Filiz AK, et al. Galium aparine L. protects against acetaminophen-induced hepatotoxicity in rats. Chem Biol Interact. 2022;366. doi:10.1016/J.CBI.2022.110119
Shi G, Liu J, Zhao W, Liu Y, Tian X. Separation and purification and in vitro anti-proliferative activity of leukemia cell K562 of Galium aparine L. petroleum ether phase. Saudi Pharm J. 2016;24(3):241-244. doi:10.1016/J.JSPS.2016.04.005
Hartwell JL. Plants used against cancer. A survey. Lloydia. 1971;34(4):386-425.
Atmaca H, Bozkurt E, Cittan M, Dilek Tepe H. Effects of Galium aparine extract on the cell viability, cell cycle and cell death in breast cancer cell lines. J Ethnopharmacol. 2016;186:305-310. doi:10.1016/j.jep.2016.04.007
Schmidt M, Scholz CJ, Gavril GL, et al. Effect of Galium verum aqueous extract on growth, motility and gene expression in drug-sensitive and -resistant laryngeal carcinoma cell lines. Int J Oncol. 2014;44(3):745-760. doi:10.3892/IJO.2013.2220

Uribe ML, Marrocco I, Yarden Y. EGFR in Cancer: Signaling Mechanisms, Drugs, and Acquired Resistance. Cancers 2021, Vol 13, Page 2748. 2021;13(11):2748. doi:10.3390/-CANCERS13112748
Albadari N, Xie Y, Li W. Deciphering treatment resistance in metastatic colorectal cancer: roles of drug transports, EGFR mutations, and HGF/c-MET signaling. Front Pharmacol. 2023;14:1340401. doi:10.3389/FPHAR.2023.1340401/BIBTEX
Citri A, Yarden Y. EGF-ERBB signalling: towards the systems level. Nat Rev Mol Cell Biol. 2006;7(7):505-516. doi:10.1038/NRM1962
Mitsudomi T, Yatabe Y. Epidermal growth factor receptor in relation to tumor development: EGFR gene and cancer. FEBS J. 2010;277(2):301-308. doi:10.1111/J.1742-4658.2009.07448.X
McKay JA, Murray LJ, Curran S, et al. Evaluation of the epidermal growth factor receptor (EGFR) in colorectal tumours and lymph node metastases. Eur J Cancer. 2002;38(17):2258-2264. doi:10.1016/S0959-8049(02)00234-4
Goldstein NS, Armin M. Epidermal Growth Factor Receptor Immunohistochemical Reactivity in Patients with American Joint Committee on Cancer Stage IV Colon Adenocarcinoma Implications for a Standardized Scoring System. Published online 2001. doi:10.1002/1097-0142
Ercan D, Xu C, Yanagita M, et al. Reactivation of ERK signaling causes resistance to EGFR Kinase inhibitors. Cancer Discov. 2012;2(10):934-947. doi:10.1158/2159-8290.CD-12-0103
Ponsioen B, Post JB, Buissant des Amorie JR, et al. Quantifying single-cell ERK dynamics in colorectal cancer organoids reveals EGFR as an amplifier of oncogenic MAPK pathway signalling. Nature Cell Biology. 2021;23(4):377-390. doi:10.1038/s41556-021-00654-5
Öztürk A, Taşkıran AŞ, Gündoğdu E. The role of oxidative stress in the protective effect of boric acid against glutamate excitotoxicity in C6 glioma cells. Journal of Boron. 2025;10(1):1-9. doi:10.30728/boron.1519354
Yulak F, Filiz AK, Joha Z, Ergul M. Mechanism of anticancer effect of ETP-45658, a PI3K/AKT/mTOR pathway inhibitor on HT-29 Cells. Medical Oncology. 2023;40(12):341. doi:10.1007/s12032-023-02221-4
Yulak F, Joha Z, Öztürk A, İnan ZDŞ, Taşkıran AŞ. Enoxaparin Protects C6 Glioma Cells from Glutamate-Induced Cytotoxicity by Reducing Oxidative Stress and Apoptosis. Mol Neurobiol. 2025:4631-4640. doi:10.1007/s12035-024-04587-6
Siegel RL, Wagle NS, Cercek A, Smith RA, Jemal A. Colorectal cancer statistics, 2023. CA Cancer J Clin. 2023;73(3):233-254. doi:10.3322/CAAC.21772
Chen Y, Cheng CS, Tan HY, Tam CW, Wang N, Feng Y. Efficacy of Herbal Medicines Intervention for Colorectal Cancer Patients With Chemotherapy-Induced Gastrointestinal Toxicity — a Systematic Review and Meta-Analysis. Front Oncol. 2021;11:629132. doi:10.3389/FONC.2021.629132/FULL
Yue GGL, Jiang L, Kwok HF, et al. Turmeric ethanolic extract possesses stronger inhibitory activities on colon tumour growth than curcumin - The importance of turmerones. J Funct Foods. 2016;22:565-577. doi:10.1016/j.jff.2016.02.011
Wang J, Zhang J, Shi Y, et al. Mechanistic Investigation of the Specific Anticancer Property of Artemisinin and Its Combination with Aminolevulinic Acid for Enhanced Anticolorectal Cancer Activity. ACS Cent Sci. 2017;3(7):743-750. doi:10.1021/ACSCENTSCI.7B00156
Schmidt M, Polednik C, Roller J, Hagen R. Galium verum aqueous extract strongly inhibits the motility of head and neck cancer cell lines and protects mucosal keratinocytes against toxic DNA damage. Oncol Rep. 2014;32(3):1296-1302. doi:10.3892/OR.2014.3316
Atmaca H. Effects of Galium aparine Extract on the Angiogenic Cytokines and ERK1/2 proteins in Human Breast Cancer Cells. Celal Bayar Üniversitesi Fen Bilimleri Dergisi. 2017;13(1):171-179. doi:10.18466/CBAYARFBE.281836
Riahi-Chebbi I, Haoues M, Essafi M, et al. Quince peel polyphenolic extract blocks human colon adenocarcinoma LS174 cell growth and potentiates 5-fluorouracil efficacy. Cancer Cell Int. 2016;16(1):1. doi:10.1186/s12935-016-0276-7
Kwok AHY, Wang Y, Ho WS. Cytotoxic and pro-oxidative effects of Imperata cylindrica aerial part ethyl acetate extract in colorectal cancer in vitro. Phytomedicine. 2016;23(5):558-565. doi:10.1016/j.phymed.2016.02.015
Napolitano S, Martini G, Ciardiello D, et al. Targeting the EGFR signalling pathway in metastatic colorectal cancer. Lancet Gastroenterol Hepatol. 2024;9(7):664-676. doi:10.1016/s2468-1253(23)00479-x
Venkateswaran S, Manivannan HP, Francis AP, Veeraraghavan VP, R G, Sankaran K. Identification of Potential Phytochemical Inhibitors From Conium maculatum Targeting the Epidermal Growth Factor Receptor in Metastatic Colorectal Cancer via Molecular Docking Analysis. Cureus. 2023;15(10). doi:10.7759/CUREUS.48000
Cohen RB. Epidermal Growth Factor Receptor as a Therapeutic Target in Colorectal Cancer. Clin Colorectal Cancer. 2003;2(4):246-251. doi:10.3816/CCC.2003.N.006
Sahyon H, Abdelhamid N, Hanafy N, et al. The Effect of Encapsulated Apigenin Nanoparticles on HePG-2 Cells through Regulation of P53. Pharmaceutics. 2022;2022:1160. doi:10.3390/pharmaceutics14061160
Pandey P, Khan F, Upadhyay TK. Deciphering the modulatory role of apigenin targeting oncogenic pathways in human cancers. Chem Biol Drug Des. 2023;101(6):1446-1458. doi:10.1111/CBDD.14206
Ahmed SA, Parama D, Daimari E, et al. Rationalizing the therapeutic potential of apigenin against cancer. Life Sci. 2021;267. doi:10.1016/j.lfs.2020.118814
Bhatiya M, Pathak S, Jothimani G, Duttaroy AK, Banerjee A. A Comprehensive Study on the Anti-cancer Effects of Quercetin and Its Epigenetic Modifications in Arresting Progression of Colon Cancer Cell Proliferation. Arch Immunol Ther Exp (Warsz). 2023;71(1):1-16. doi:10.1007/S00005-023-00669-W/FIGURES/7
Zhang XA, Zhang S, Yin Q, Zhang J. Quercetin induces human colon cancer cells apoptosis by inhibiting the nuclear factor-kappa B Pathway. Pharmacogn Mag. 2015;11(42):404. doi:10.4103/0973-1296.153096
Yoo HS, Won SB, Kwon YH. Luteolin Induces Apoptosis and Autophagy in HCT116 Colon Cancer Cells via p53-Dependent Pathway. Nutr Cancer. 2022;74(2):677-686. doi:10.1080/01635581.2021.1903947
Ranjbary AG, Bagherzadeh A, Sabbaghi SS, et al. Chlorogenic acid induces apoptosis and cell-cycle arrest in colorectal cancer cells. Mol Biol Rep. 2023;50(12):9845-9857. doi:10.1007/S11033-023-08854-Y

Details

Primary Language

English

Subjects

Medical Physiology (Other)

Journal Section

Research Article

Authors

Ayşegül Öztürk ^*
0000-0001-8130-7968
Türkiye

Arzuhan Çetindağ
0000-0002-5420-3546
Türkiye

Fatih Yulak
0000-0003-3708-6752
Türkiye

Sebahattin Karabulut
0000-0002-3261-4125
Türkiye

Ahmet Kemal Filiz
0000-0001-9260-5549
Türkiye

Publication Date

September 27, 2026

Submission Date

March 10, 2026

Acceptance Date

June 16, 2026

Published in Issue

Year 2026 Volume: 6 Number: 3

DOI

https://doi.org/10.66235/kumj.1906992

IZ

https://izlik.org/JA44ZS86LN

Cite

RIS / Bibtex

APA

Öztürk, A., Çetindağ, A., Yulak, F., Karabulut, S., & Filiz, A. K. (2026). Galium aparine L. extract inhibits the proliferation of HT-29 cells and induces apoptosis. Kastamonu Medical Journal, 6(3), 254-262. https://doi.org/10.66235/kumj.1906992

AMA

1.Öztürk A, Çetindağ A, Yulak F, Karabulut S, Filiz AK. Galium aparine L. extract inhibits the proliferation of HT-29 cells and induces apoptosis. Kastamonu Medical Journal. 2026;6(3):254-262. doi:10.66235/kumj.1906992

Chicago

Öztürk, Ayşegül, Arzuhan Çetindağ, Fatih Yulak, Sebahattin Karabulut, and Ahmet Kemal Filiz. 2026. “Galium Aparine L. Extract Inhibits the Proliferation of HT-29 Cells and Induces Apoptosis”. Kastamonu Medical Journal 6 (3): 254-62. https://doi.org/10.66235/kumj.1906992.

EndNote

Öztürk A, Çetindağ A, Yulak F, Karabulut S, Filiz AK (September 1, 2026) Galium aparine L. extract inhibits the proliferation of HT-29 cells and induces apoptosis. Kastamonu Medical Journal 6 3 254–262.

IEEE

[1]A. Öztürk, A. Çetindağ, F. Yulak, S. Karabulut, and A. K. Filiz, “Galium aparine L. extract inhibits the proliferation of HT-29 cells and induces apoptosis”, Kastamonu Medical Journal, vol. 6, no. 3, pp. 254–262, Sept. 2026, doi: 10.66235/kumj.1906992.

ISNAD

Öztürk, Ayşegül - Çetindağ, Arzuhan - Yulak, Fatih - Karabulut, Sebahattin - Filiz, Ahmet Kemal. “Galium Aparine L. Extract Inhibits the Proliferation of HT-29 Cells and Induces Apoptosis”. Kastamonu Medical Journal 6/3 (September 1, 2026): 254-262. https://doi.org/10.66235/kumj.1906992.

JAMA

1.Öztürk A, Çetindağ A, Yulak F, Karabulut S, Filiz AK. Galium aparine L. extract inhibits the proliferation of HT-29 cells and induces apoptosis. Kastamonu Medical Journal. 2026;6:254–262.

MLA

Öztürk, Ayşegül, et al. “Galium Aparine L. Extract Inhibits the Proliferation of HT-29 Cells and Induces Apoptosis”. Kastamonu Medical Journal, vol. 6, no. 3, Sept. 2026, pp. 254-62, doi:10.66235/kumj.1906992.

Vancouver

1.Ayşegül Öztürk, Arzuhan Çetindağ, Fatih Yulak, Sebahattin Karabulut, Ahmet Kemal Filiz. Galium aparine L. extract inhibits the proliferation of HT-29 cells and induces apoptosis. Kastamonu Medical Journal. 2026 Sep. 1;6(3):254-62. doi:10.66235/kumj.1906992