In vitro and in silico evaluation of Acalypha indica ethanolic extract on lipid metabolism-related protein expression on breast cancer cells
DOI:
https://doi.org/10.11113/jomalisc.v3.74Keywords:
Acalypha indica, fatty acid synthase, acetyl-CoA carboxylase, ATP-citrate lyase, breast cancerAbstract
Dysregulated lipid metabolism has been recognized as one of the most significant metabolic changes in breast cancer development. Acalypha indica is a polyphenol-rich herbal plant with various medicinal applications such as antibacterial, antioxidant, and anticancer activities. However, studies on the inhibitory effect of A. indica ethanolic extract (AI) on breast cancer cells and its efficacy as an alternative lipogenesis inhibitor are still less explored. Therefore, this study was designed to investigate the possible inhibition mechanisms of selected phenolic acids and flavonoids from the extract against MCF-7 breast cancer. The antiproliferative effect of AI was determined using MTT assay. Immunocytochemical analysis was used to evaluate the inhibitory effect of AI on the expression of the three lipogenic enzymes; fatty acid synthase (FASN), acetyl-CoA carboxylase (ACCA), and ATP citrate lyase (ACLY). In addition, molecular docking of the phenolic acids (gallic acid, ferulic acid, and caffeic acid) and flavonoids (rutin and quercetin) were performed using AutoDock Tools, and their ADMET properties were determined using SwissADME and pKCSM web servers. AI treatment resulted in inhibition of cell viability of MCF-7 breast cancer cells after 24 and 48 h. In addition, immunocytochemistry revealed a significant decrease in the expression of FASN, ACCA, and ACLY in MCF-7 cells when compared to control. Docking studies demonstrated that AI phytochemical quercetin was considered the most potential lipogenesis inhibitor due to its lowest binding energy to FASN, ACCA, and ACLY compared to other phytochemicals. Nevertheless, results from ADMET analysis suggested that all compounds except rutin possessed acceptable ADME and toxicity properties. Collectively, these findings indicated that the chemoprevention by AI in breast cancer cells is associated with the inhibition of fatty acid synthesis.
References
Heer, E., Harper, A., Escandor, N., Sung, H., McCormack, V., & Fidler-Benaoudia, M. M. (2020). Global burden and trends in premenopausal and postmenopausal breast cancer: A population-based study. The Lancet Global Health, 8(9), e1027–e1037.
Schmidt, H., & Barnhill, A. (2015). Equity and noncommunicable disease reduction under the Sustainable Development Goals. PLoS Medicine, 12(9), e1001872.
Butler, L. M., Perone, Y., Dehairs, J., Lupien, L. E., de Laat, V., Talebi, A., ... & Swinnen, J. V. (2020). Lipids and cancer: Emerging roles in pathogenesis, diagnosis, and therapeutic intervention. Advanced Drug Delivery Reviews, 159, 245–293.
Kuhajda, F. P. (2006). Fatty acid synthase and cancer: New application of an old pathway. Cancer Research, 66(12), 5977–5980.
Blücher, C., & Stadler, S. C. (2017). Obesity and breast cancer: Current insights on the role of fatty acids and lipid metabolism in promoting breast cancer growth and progression. Frontiers in Endocrinology, 8, 293
Lee, K., Kruper, L., Dieli-Conwright, C. M., & Mortimer, J. E. (2019). The impact of obesity on breast cancer diagnosis and treatment. Current Oncology Reports, 21(5), 41.
Lee, J. S., Sul, J. Y., Park, J. B., Lee, M. S., Cha, E. Y., Song, I. S., ... & Chang, E. S. (2012). Fatty acid synthase inhibition by amentoflavone suppresses HER2/neu (erbB2) oncogene in SKBR3 human breast cancer cells. Phytotherapy Research, 27(5), 713–720.
Zhu, W., Lv, C., Wang, J., Gao, Q., Zhu, H., & Wen, H. (2017). Patuletin induces apoptosis of human breast cancer SK-BR-3 cell line via inhibiting fatty acid synthase gene expression and activity. Oncology Letters, 14(6), 7449–7454.
Wang, Y., Nie, F., Ouyang, J., Wang, X., & Ma, X. (2014). Inhibitory effects of sea buckthorn procyanidins on fatty acid synthase and MDA-MB-231 cells. Tumor Biology, 35(10), 9563–9569.
Lee, J. S., Lee, M. S., Oh, W. K., & Sul, J. Y. (2009). Fatty acid synthase inhibition by amentoflavone induces apoptosis and antiproliferation in human breast cancer cells. Biological and Pharmaceutical Bulletin, 32(8), 1427–1432.
Wang, D., Yin, L., Wei, J., Yang, Z., & Jiang, G. (2017). ATP citrate lyase is increased in human breast cancer, depletion of which promotes apoptosis. Tumor Biology, 39(4), 1–10.
Ismail, A., Mokhlis, H. A., Sharaky, M., Sobhy, M. H., Hassanein, S. S., Doghish, A. S., ... & Attia, Y. M. (2022). Hydroxycitric acid reverses tamoxifen resistance through inhibition of ATP citrate lyase. Pathology Research and Practice, 240, 154188.
Yoon, S., Lee, M. Y., Park, S. W., Moon, J. S., Koh, Y. K., Ahn, Y. H., ... & Kim, K. S. (2007). Up-regulation of acetyl-CoA carboxylase α and fatty acid synthase by human epidermal growth factor receptor 2 at the translational level in breast cancer cells. The Journal of Biological Chemistry, 282(36), 26122–26131.
Liu, H., Liu, Y., & Zhang, J. T. (2008). A new mechanism of drug resistance in breast cancer cells: Fatty acid synthase overexpression-mediated palmitate overproduction. Molecular Cancer Therapeutics, 7(2), 263–270.
Chekuri, S., Vankudothu, N., Panjala, S., Rao, N. B., & Anupalli, R. R. (2016). Phytochemical analysis, antioxidant and antimicrobial activity of Acalypha indica leaf extracts in different organic solvents. International Journal of Phytomedicine, 8(3), 444–452.
Chekuri, S., Panjala, S., & Anupalli, R. R. (2017). Cytotoxic activity of Acalypha indica L. hexane extract on breast cancer cell lines (MCF-7). The Journal of Phytopharmacology, 6(5), 264–268.
Ibrahim, A. M., Hamid, M. A., Althiab, R. A., Shariff, A. H. M., & Zulkifli, R. M. (2021). In vitro fibroblasts viability and migration stimulation of Acalypha indica: An insight on wound healing activity. Future Journal of Pharmaceutical Sciences, 7(1), 1–6.
Priya, C. L., & Rao, K. V. B. (2014). Acute and sub-chronic toxicological studies on methanolic stem extract of Acalypha indica Linn in albino Wistar rats. International Journal of Pharmacy and Pharmaceutical Science, 6(9), 560–563.
Hazali, N., Mohd Nazri, N. N., Ibrahim, M., & Masri, M. (2016). Subchronic toxicity of Malaysian Acalypha indica: Biochemistry and haematology analysis of rat. Jurnal Teknologi, 78(5), 21–26.
Kavitha, S., Kalai Kovan, T., & Vijaya Bharathi, R. (2009). In vitro antioxidant and anticancer studies on the leaf of Acalypha indica. Biomedical & Pharmacology Journal, 2(2), 431–435.
Maulidina, A. A., & Farida, S. (2019). Treatment of fatty pancreas: Acalypha indica Linn. extract as an alternative to simvastatin. AIP Conference Proceedings, 2193(030025), 1–5.
Masih, M., Banerjee, T., Banerjee, B., & Pal, A. (2011). Antidiabetic activity of Acalypha indica Linn. on normal and alloxan-induced diabetic rats. International Journal of Pharmacy and Pharmaceutical Science, 3(3), 51–54.
Rajasekaran, S., Anandan, R., & Nishad, K. M. (2013). Anti-hyperlipidemic activity of Acalypha indica Linn. on induced hyperlipidemia. International Journal of Pharmacy and Pharmaceutical Science, 5(4), 699–701.
Brusselmans, K., Vrolix, R., Verhoeven, G., & Swinnen, J. V. (2005). Induction of cancer cell apoptosis by flavonoids is associated with their ability to inhibit fatty acid synthase activity. Journal of Biological Chemistry, 280(7), 5636–5645.
Jun, C., Donghong, Z., Cai, W., Xu, L., Li, E., Wu, Y., & Kazuo, S. (2009). Inhibitory effects of four plant flavonoid extracts on fatty acid synthase. Journal of Environmental Sciences, 21, 131–134.
Sun, W., Shahrajabian, M. H., & Cheng, Q. (2021). Natural dietary and medicinal plants with anti-obesity therapeutic activities for treatment and prevention of obesity during lockdown and in the post-COVID-19 era. Applied Sciences, 11(2889), 1–23.
Wang, Y., Li, J. Y., Han, M., Wang, W. L., & Li, Y. Z. (2015). Prevention and treatment effect of total flavonoids in Stellera chamaejasme L. on nonalcoholic fatty liver in rats. Lipids in Health and Disease, 14(85), 1–9.
Liu, H., Huang, X., & Ye, T. (2018). MiR-22 down-regulates the proto-oncogene ATP citrate lyase to inhibit the growth and metastasis of breast cancer. American Journal of Translational Research, 10(3), 659–669.
Godipurge, S. S., Biradar, J. S., & Mahurkar, N. (2014). Phytochemical and pharmacological evaluation of Acalypha indica Linn in experimental animal models. International Journal of Pharmacognosy and Phytochemical Research, 6(4), 973–979.
Kumar, P., Chakrapani, P., Arunjyothi, B., & Rojarani, A. (2016). Anti-epileptic activity of Acalypha indica methanolic leaves extract with animal experiment. International Journal of Pharmacognosy and Phytochemical Research, 8(9), 1560–1565.
Pemble, C. W., Johnson, L. C., Kridel, S. J., & Lowther, W. T. (2007). Crystal structure of the thioesterase domain of human fatty acid synthase inhibited by Orlistat. Nature Structural & Molecular Biology, 14(8), 704–709.
Griffith, D. A., Kung, D. W., Esler, W. P., Amor, P. A., Bagley, S. W., Beysen, C., ... & Green, M. (2014). Decreasing the rate of metabolic ketone reduction in the discovery of a clinical acetyl-CoA carboxylase inhibitor for the treatment of diabetes. Journal of Medicinal Chemistry, 57(24), 10512–10526.
Huang, T., Sun, J., Wang, Q., Gao, J., & Liu, Y. (2015). Synthesis, biological evaluation, and molecular docking studies of piperidinylpiperidines and spirochromanones possessing quinoline moieties as acetyl-CoA carboxylase inhibitors. Molecules, 20(9), 16221–16234.
Vyas, V. K., Dabasia, M., Qureshi, G., & Patel, P. (2016). Molecular modeling study for the design of novel acetyl-CoA carboxylase inhibitors using 3D QSAR, molecular docking, and dynamic simulations. Journal of Biomolecular Structure and Dynamics, 34(11), 2423–2441.
Ismail, A., Doghish, A. S., Elsadek, B. E. M., Salama, S. A., & Mariee, A. D. (2020). Hydroxycitric acid potentiates the cytotoxic effect of tamoxifen in MCF-7 breast cancer cells through inhibition of ATP citrate lyase. Steroids, 160, 108643.
Lipinski, C. A., Lombardo, F., Dominy, B. W., & Feeney, P. J. (2001). Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings. Advanced Drug Delivery Reviews, 46(1–3), 3–26.
