Effects of Curcumin and Achillea millefolium L. Extract on the Expression of BCL2, DLEC1, MCC, and NANOG Genes in LS180 Colorectal Cancer Cell Line

Articles in Press

Document Type : Research Article

Authors

Department of Biology, IL.C., Islamic Azad University, Ilam, Iran

Abstract

Colorectal cancer is a common gastrointestinal malignancy with high incidence and mortality rates. Due to increasing resistance to conventional chemotherapy, identifying effective natural compounds has become essential. This study investigated the independent and combined effects of Achillea millefolium L. (Yarrow) extract and curcumin on cell viability and the expression of DLEC1, MCC, BCL2, and NANOG genes in LS180 colorectal cancer cells. LS180 cells were treated with various concentrations of yarrow extract (2.5, 5, 10, 20, and 40 μg/mL), curcumin (6.25, 12.5, 25, 50, and 75 μg/mL), and their combination for 24 and 48 hours. Cell viability was assessed using the MTT assay, and gene expression was measured by qRT-PCR. Statistical analyses were performed using Tukey's test, with a p< 0.05 considered statistically significant. Both yarrow extract and curcumin reduced LS180 cell viability in a dose- and time-dependent manner. The combined treatment exhibited a strong synergistic effect in inhibiting cancer cell proliferation. Furthermore, the combination significantly increased the expression of the tumor suppressor genes DLEC1 (22.29- fold) and MCC (20.96- fold) compared to the control group (p< 0.0001). All three treatments also led to a significant reduction in the expression of the oncogenes BCL2 and NANOG. The greatest reduction was observed following curcumin treatment alone, with a 0.069- fold decrease (equivalent to 93.12%) in BCL2 and a 0.160- fold decrease (equivalent to 84.01%) in NANOG expression (p< 0.0001). The combination of yarrow extract and curcumin, by markedly upregulating tumor suppressor genes, presents an effective synergistic strategy for inhibiting LS180 cancer cells. Although the combined treatment was only slightly less effective than curcumin alone in reducing BCL2 and NANOG expression, it holds considerable promise for combinatorial applications in colorectal cancer therapy due to its stronger induction of tumor suppression pathways and synergistic cytotoxicity.

Keywords

Main Subjects

References

Ahmad, I., Ahmad, S., Samad, M. A., Adam, A. M., Zughaibi, T. A., Alhosin, M., ... & Tabrez, S. (2025). Synergistic inhibition of colon cancer cell proliferation via p53, Bax, and Bcl2 modulation by curcumin and plumbagin combination. American Chemical Society Omega, 10(18), 19045-19060. https://doi.org/10.1021/acsomega.5c01258
Amaroli, A., Panfoli, I., Bozzo, M., Ferrando, S., Candiani, S., & Ravera, S. (2024). The bright side of curcumin: A narrative review of its therapeutic potential in cancer management. Cancers, 16(14), 2580. https://doi.org/10.3390/cancers16142580
Aromokeye, R., & Si, H. (2022). Combined curcumin and luteolin synergistically inhibit colon cancer associated with notch1 and TGFβ signaling pathways in cultured cells and xenograft mice. Cancers, 14(12), 3001.‏https://doi.org/10.3390/cancers14123001
Brancato, D., Bruno, F., Coniglio, E., Sturiale, V., Saccone, S., & Federico, C. (2024). The chromatin organization close to SNP rs12913832, involved in eye color variation, is evolutionary conserved in vertebrates. International Journal of Molecular Sciences, 25(12), 6602.‏ https://doi.org/10.3390/ijms25042377
Burgman, B., Pan, X., Agrawal, A., Sahni, N., & Yi, S. S. (2024). Abstract LB251: Widespread epigenetic deregulation and its dependency on tumor sidedness in colorectal cancer. Cancer Research, 84(7), LB251-LB251. https://doi.org/10.1158/1538-7445.am2024-lb251
Chikaishi, Y., Matsuoka, H., Sugihara, E., Takeda, M., Sumitomo, M., Yamada, S., ... & Takimoto, T. (2025). Mutation analysis of TMB‐High colorectal cancer: Insights into molecular pathways and clinical implications. Cancer Science, 116(4), 1082-1093. ‏https://doi.org/10.1111/cas.16455
Deng, L., Zhang, X., Xiang, X., Xiong, R., Xiao, D., Chen, Z., ... & Feng, G. (2021). NANOG promotes cell proliferation, invasion, and stemness via IL-6/STAT3 signaling in esophageal squamous carcinoma. Technology in Cancer Research and Treatment, 20, 15330338211038492.‏ https://doi.org/10.1177/15330338211038492
Duta-Ion, S. G., Juganaru, I. R., Hotinceanu, I. A., Dan, A., Burtavel, L. M., Coman, M. C., ... & Radoi, V. E. (2024). Redefining therapeutic approaches in colorectal cancer: targeting molecular pathways and overcoming resistance. International Journal of Molecular Sciences, 25(23), 12507. ‏https://doi.org/10.3390/ijms252312507
Far, B. F., Behzad, G., & Khalili, H. (2023). Achillea millefolium: Mechanism of action, pharmacokinetic, clinical drug-drug interactions and tolerability. Heliyon, 9(12). https://doi.org/10.1016/j.heliyon.2023.e22841
Farooqui, A. A., & Farooqui, T. (2013). Is diabetes a risk factor for Parkinson Disease? Metabolic Syndrome and Neurological Disorders, 327-334. https://doi.org/10.1002/9781118395318
Jeter, C. R., Yang, T., Wang, J., Chao, H. P., & Tang, D. G. (2015). Concise review: NANOG in cancer stem cells and tumor development: an update and outstanding questions. Stem Cells, 33(8), 2381-2390. https://doi.org/10.1002/stem.2007
Kanwal, R., Datt, M., Liu, X., & Gupta, S. (2016). Dietary flavones as dual inhibitors of DNA methyltransferases and histone methyltransferases. Public Library of Science ONE, 11(9), e0162956.‏ https://doi.org/10.1371/journal.pone.0162956
Kanwal, R., Yang, X., Shankar, E., & Gupta, S. (2017). Luteolin selectively inhibits EZH2 and blocks H3K27 methylation in prostate cancer cells. Cancer Research, 77(13), 2230-2230. https://doi.org/10.1158/1538-7445.AM2017-2230
Lee, S., Lee, B., Kwon, S. H., Park, J., & Kim, S. H. (2025). MCC in the spotlight: Its dual role in signal regulation and oncogenesis. Cellular Signalling, 131, 111756. https://doi.org/10.1016/j.cellsig.2025.111756
Najafzadeh, B., Asadzadeh, Z., Motafakker Azad, R., Mokhtarzadeh, A., Baghbanzadeh, A., Alemohammad, H., ... & Baradaran, B. (2021). The oncogenic potential of NANOG: An important cancer induction mediator. Journal of Cellular Physiology, 236(4), 2443-2458. https://doi.org/10.1002/jcp.30063
Nguyen, H., Huang, Q., Juang, U., Gwon, S., Jung, W., Lee, S., ... & Kim, S. H. (2024). The mutated in colorectal cancer (MCC) gene can serve as a potential biomarker of glioblastoma. Frontiers in Oncology, 14, 1435605.‏ https://doi.org/10.3389/fonc.2024.1435605
Okitsu, Y., Nagano, M., Yamagata, T., Ito, C., Toshimori, K., Dohra, H., ... & Yogo, K. (2020). Dlec1 is required for spermatogenesis and male fertility in mice. Scientific Reports, 10(1), 18883.‏ https://doi.org/10.1038/s41598-020-75957-y
Pangon, L., Mladenova, D., Watkins, L., Van Kralingen, C., Currey, N., Al‐Sohaily, S., ... & Kohonen‐Corish, M. R. (2015). MCC inhibits beta‐catenin transcriptional activity by sequestering DBC1 in the cytoplasm. International Journal of Cancer, 136(1), 55-64.‏ https://doi.org/10.1002/ijc.28967
Ravindranathan, P., Pasham, D., Balaji, U., Cardenas, J., Gu, J., Toden, S., & Goel, A. (2018). A combination of curcumin and oligomeric proanthocyanidins offer superior anti-tumorigenic properties in colorectal cancer. Scientific Reports, 8(1), 13869.‏ https://doi.org/10.1038/s41598-018-32267-8
shehzad, A., Lee, J., & Lee, Y. S. (2013). Curcumin in various cancers. Biofactors, 39(1), 56-68. https://doi.org/10.1002/biof.1068
Ying, J., Poon, F., Yu, J., Geng, H., Wong, A. H. Y., Qiu, G. H., ... & Tao, Q. (2009). DLEC1 is a functional 3p22. 3 tumor suppressors silenced by promoter CpG methylation in colon and gastric cancers. British Journal of Cancer, 100(4), 663-669.‏ https://doi.org/10.1038/sj.bjc.6604888
Yuan, Z., Hu, J., Cai, C., Liu, F., Chen, H., & Zeng, B. (2026). Integrated network pharmacology and experimental verification to reveal the mechanisms of curcumin in the treatment of colorectal cancer. Frontiers in Pharmacology, 16, 1703562. https://doi.org/10.3389/fphar.2025.1703562
Zhang, C., Zhao, Y., Yang, Y., Zhong, C., Ji, T., Duan, J., & Wang, Y. (2021). RNAi mediated silencing of NANOG expression suppresses the growth of human colorectal cancer stem cells. Biochemical and Biophysical Research Communications, 534, 254-260. https://doi.org/10.1016/j.bbrc.2020.11.101
 
Journal of Genetic Resources

Articles in Press, Accepted Manuscript
Available Online from 10 July 2026

Files

Share

How to cite

Statistics