Citrus Flavonoids from Citrus reticulata Peels Potentially Target an Autophagy Modulator, MAP1LC3A, in Breast Cancer

Bayu Anggoro, Dennaya Kumara, Dhella Angelina, Muthi Ikawati


Citrus flavonoids have been known for their vast biological activities including chemoprevention activities. However, the organic solvent extraction system limits its potential utilization. We recently adopted a hydrodynamic-cavitation method to extract citrus flavonoids from citrus peels. In this study we verified the high flavonoid content of the hydrodynamic-cavitation extract from Citrus reticulata peels and explore the potency of its citrus flavonoid contents as targeted chemoprevention agent for breast cancer by using bioinformatics. Based on a thin layer chromatography, the extract positively yielded high content of citrus flavonoids represented by hesperidin. The toxicity analysis by Protox II Online Tool revealed that hesperidin as the major citrus flavonoid in the extract was considered safe with a predicted LD50 of 12,000 mg/kg. We then further exploring citrus flavonoids’ capacity in targeting MAP1LC3A, a key protein in autophagy. UALCAN analysis validated that low expression of MAP1LC3A is associated with low survival rates in breast cancer patients. Limonin, hesperidin, narirutin, neohesperidine, and naringin are flavonoids from citrus peels that predicted to have inhibitory activity against Protein Kinase A (PKA), a negative upstream of MAP1LC3A, calculated by KNIME. Citrus flavonoids scoparone, cirsimaritin, 4',5,7-trimethoxyflavone, eupatorine, and hesperidin were also exhibit similar structure to an agonist of ATG4B, a protein that plays a role in MAP1LC3A activation. Furthermore, eupatorine, hesperidin, and cirsimaritin displayed a high affinity to ATG4B based on a molecular docking. We concluded that citrus flavonoids from citrus peels are safe to normal cells, and the citrus flavonoids potentially targets MAP1LC3A by inhibiting PKA and acting as ATG4B agonists. Thus, this extract-contained flavonoids from citrus peels is potential to be investigated further as a chemoprevention agent by inducing autophagy, especially for breast cancer.

Keywords: Citrus reticulata, citrus flavonoid, autophagy, MAP1LC3A, breast cancer.

Full Text:



Artun, F.T., Karagoz, A., Ozcan, G., Melikoglu, G., Anil, S., Kultur, S., and Sutlupinar, N., 2016, In vitro anticancer and cytotoxic activities of some plant extracts on HeLa and Vero cell lines, Journal of B.U.ON., 21(3), 720-725. CrossRef

Banerjee, P., Eckert, A.O., Schrey A.K., and Preissner, R., 2018, ProTox-II: a webserver for the prediction of toxicity of chemicals, Nucleic Acids Research, 46(W1), W257-W263. CrossRef

Bonam, S.R., Bayry, J., Tschan, M.P., and Muller, S., 2020, Progress and Challenges in The Use of MAP1LC3 as a Legitimate Marker for Measuring Dynamic Autophagy In Vivo, Cells, 9(5), 1321. CrossRef

Cherra, S.J., Kulich, S.M., Uechi, G., Balasubramani, M., Mountzouris, J., Day, B.W., et al., 2010, Regulation of the autophagy protein LC3 by phosphorylation, The Journal of Cell Biology, 190(4), 533-539. CrossRef

Chandrashekar, D.S., Bashel, B., Balasubramanya, S.A.H., Creighton, C.J., Ponce-Rodriguez, I., Chakravarthi, B.V.S.K., et al., 2017, UALCAN: A Portal for Facilitating Tumor Subgroup Gene Expression and Survival Analyses, Neoplasia, 19(8), 649-658. CrossRef

Costa, J.R., Prak, K., Aldous, S., Gewinner, C.A., and Ketteler, R., 2016, Autophagy gene expression profiling identifies a defective microtubule-associated protein light chain 3A mutant in cancer, Oncotarget, 7(27), 41203-41216. CrossRef

Goh, J.X.H., Tan, L.T.-H., Goh, J.K., Chan, K.G., Pusparajah, P., Lee, L.-H., and Goh, B.-H., 2019, Nobiletin and Derivatives: Functional Compounds from Citrus Fruit Peel for Colon Cancer Chemoprevention, Cancers, 11(6), 867. CrossRef

He, H., Dang, Y., Dai, F., Guo, Z., Wu, J., She, X., et al., 2003, Post-translational modifications of three members of the human MAP1LC3 family and detection of a novel type of modification for MAP1LC3B, The Journal of Biological Chemistry, 278(31), 29278–29287. CrossRef

Hermawan, A., Putri, H., and Utomo, R.Y., 2020, Functional network analysis reveals potential repurposing of β-blocker atenolol for pancreatic cancer therapy, DARU Journal of Pharmaceutical Sciences, 28(2), 685-699. CrossRef

Ikawati, M., Armandari, I., Khumaira, A., and Ertanto, Y., 2019, Effect of peel extract from Citrus reticulata and hesperidin, a citrus flavonoid, on macrophage cell line, Indonesian Journal of Pharmacy, 30(4), 260-268. CrossRef

Jenie, R.I., Handayani, S., Susidarti, R.A., Udin, L.Z., and Meiyanto, E., 2018, The Cytotoxic and Antimigratory Activity of Brazilin-Doxorubicin on MCF-7/HER2 Cells, Advanced Pharmaceutical Bulletin, 8(3), 507–516. CrossRef

Koolaji, N., Shammugasamy, B., Schindeler, A., Dong, Q., Dehghani, F., and Valtchev, P., 2020, Citrus Peel Flavonoids as Potential Cancer Prevention Agents, Current Developments in Nutrition, 4(5), nzaa025. CrossRef

Li, X., He, S., and Ma, B., 2020, Autophagy and autophagy-related proteins in cancer, Molecular Cancer, 19(1), 12. CrossRef

Meneguzzo, F., Ciriminna, R., Zabini, F., and Pagliaro, M., 2020, Review of Evidence Available on Hesperidin-Rich Products as Potential Tools against COVID-19 and Hydrodynamic Cavitation-Based Extraction as a Method of Increasing Their Production, Processes, 8(5), 2227-9717. CrossRef

Nair S,A., Sr, R.K., Nair, A.S., and Baby, S., 2018, Citrus peels prevent cancer, Phytomedicine: International Journal of Phytotherapy and Phytopharmacology, 50, 231–237. CrossRef

Othman, E.Q.G., Kaur, G., Mutee, A.F., Muhammad, T.S.T., and Tan, M.L., 2009, Immunohistochemical expression of MAP1LC3A and MAP1LC3B protein in breast carcinoma tissues, Journal of Clinical Laboratory Analysis, 23(4), 249-258. CrossRef

Utomo, R.Y., Ikawati, M., Putri, D.D.P., Salsabila, I.A., and Meiyanto, E., 2020, The Chemopreventive Potential of Diosmin and Hesperidin for COVID-19 and Its Comorbid Diseases, Indonesian Journal of Cancer Chemoprevention, 11(3), 154-167. CrossRef

World Health Organization International Agency for Research on Cancer (IARC), 2020, GLOBOCAN 2020: estimated cancer incidence, mortality and prevalence worldwide in 2020. URL:, accessed on 19 February 2021.

Zhang, L., Guo, M., Li, J., Zheng, Y., Zhang, S., Xie, T., et al., 2016, Systems biology-based discovery of a potential Atg4B agonist (Flubendazole) that induces autophagy in breast cancer, Molecular BioSystems, 11(11), 2860-28686. CrossRef


Copyright (c) 2022 Bayu Anggoro, Dennaya Kumara, Dhella Angelina, Muthi Ikawati

Indexed by:




Indonesian Society for Cancer Chemoprevention