Effects of ethanolic leaf extract of Gynura divaricata on NF-κB pathway in human HeLa cells
Keywords:
Antioxidants, Cervical cancer, Gynura divaricata, HeLa cell, Nuclear factor (NF)-kBAbstract
This study aims to evaluate the anti-cancer activity of the ethanolic leaf extract of Gynura divaricata in human cervical cancer HeLa cells. Cells were treated with various concentrations of extract, and cell viability was assessed to determine the half-maximal inhibitory concentration (IC50). The extract significantly reduced HeLa cell viability in a concentration-dependent manner, demonstrating marked cytotoxic activity. Treatment with 2x IC50 extract markedly suppressed cellular metabolic activity and significantly downregulated the expression of NF-κB-related genes compared with control and DMSO-treated cells. The extract also reduced the transcription of inflammatory and apoptosis-associated factors, including TNF-α, Bcl-2, Bax, COX-2, IKKα, IκBα, IL-8, IL-6, and IL-1β. These findings indicate that G. divaricata inhibits HeLa cell growth by suppressing NF-κB-mediated signaling and inflammatory responses, suggesting its potential as a natural therapeutic agent against cervical cancer.
0
References
Arora D, Bhalata SC. Melatonin and nitric oxide regulate sunflower seedling growth under salt stress accompanying differential expression of Cu/Zn SOD and Mn SOD. Free Radic Biol Med. 2017; 106; 315-28.
Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72; 248-54.
Chao PY, Huang WY, Hu SP, Lo HF, Lin KK, Huang MY, Chang TR, Yang CM. Indigenous purple vegetable extracts protect against hydrogen peroxide -induced DNA damage in human lymphocytes. Food Nutr Sci. 2013a; 4; 62-70.
Chao PY, Huang WY, Hu SP, Lo HF. Lin KK, Huang MY, Chang TR, Yang CM. inhibitive effects of mulberry leaf-related extracts on cell adhesion and inflammatory response in human aortic endothelial cells. Evid Based Complement Alternat Med. 2013b; 1-14.
Chao PY, Liu YF, Lin KH, Lin SY, Hsu JI, Yang CM, Lai JY. Antioxidant activity in. extracts of 27 indigenous Taiwanese vegetables. Nutrients 2014; 6; 2115-30.
Chen CM, Li SC, Lin YL, Hsu CY, Shieh MJ, Liu JF. Consump-tion of purple sweet potato leaves modulates human immune response: T-lymphocyte functions, lytic activity of natural killer cell and antibody production. World J Gastroenterol. 2005; 11; 5777-81.
Chen J, Mangelinckx S, Ma L, Wang Z, Li W, Kimpe NDe. Caffeoylquinic acid derivatives isolated from the aerial parts of Gynura divaricata and their yeast alpha glucosidase and PTP1B inhibitory activity. Fitoterapia 2014; 99; 1-6.
Chou SC, Chuang LM, Lee SS. Hypoglycemic constituents of Gynura divaricata subsp. Formosana. Nat Prod Commun. 2012; 7; 221-22.
Chung KM, Yu SW. Interplay between autophagy and programmed cell death in mammalian neural stem cells. BMB Rep. 2013; 46; 383-90.
Cruceriu D, Baldasici O, Balacescu O, Berindan-Neagoe I. The dual role of tumor necrosis factor-alpha (TNF-α) in breast cancer: Molecular insights and therapeutic approaches. Cell Oncol. 2020; 43; 1-18.
Dantu AS, Shankarguru P, Devi DR. Evaluation of in vitro anticancer activity of hydroalcoholic extract of Tabernaemontana divaricata. Asian J Pharm Clin Res. 2012; 5; 59-61.
Dhindsa RA, Plumb-Dhindsa P, Thorpe TA. Leaf senescence: Correlated with increased permeability and lipid peroxidation, and decreased levels superoxide dismutase and cata-lase. J Exp Bot. 1981; 126; 93-101.
Dong X, Zhao SX, Xu BQ, Zhang YQ. Gynura divaricata ameliorates hepatic insulin resistance by modulating insulin signalling, maintaining glycolipid homeostasis and reducing inflammation in type 2 diabetic mice. Toxicol Res. 2019; 8; 928-38.
Feng X, Shan R, Xiaomeng H. The linkage of NF‑κB signaling pathway‑associated long non‑coding RNAs with tumor microenvironment and prognosis in cervical cancer. BMC Med Genomics. 2023; 16; 169.
Foyer CH, Lopez‐Delgado H, Dat JF, Scott IM. Hydrogen peroxide and glutathione associated mechanisms of acclamatory stress tolerance and signaling. Physiol Plant. 1997; 100; 241-54.
Gansukh E, Mya KK, Jung M, Keum YS, Kim DH, Saini RK. Lutein derived from marigold (Tagetes erecta) petals triggers ROS generation and activates Bax and caspase-3 mediated apoptosis of human cervical carcinoma (HeLa) cells. Food Chem Toxicol. 2019; 127; 11-18.
Garcia-Pineres AJ, Hildesheim A, Dodd L, Kemp TJ, Yang J, Fullmer B, Pinto LA. Gene expression patterns induced by HPV-16 L1 virus-like particles in leukocytes from vaccine recipients. J Immunol. 2009; 182; 1706-29.
Gugnoni M, Sancisi V, Manzotti G, Gandolfi G, Ciarrocchi A. Autophagy and epithelial–mesenchymal transition: An intricate interplay in cancer. Cell Death Dis. 2016; 7; e2520.
Hayden MS, Ghosh S.NF-κB in immunobiology. Cell Res. 2011; 21; 223-44.
He L, Xu X, Liu A, Sun J, Xue Y. Advances in studies on Gynura divarica (L.) DC. J Med Pharm Chin Minorities. 2014; 10; 61-65.
Hong MH, Jin X J, Yoon JJ, Lee YJ, Oh HC, Lee HS, Kim HY, Kang DG. Antihypertensive effects of Gynura divaricata (L.) DC in rats with renovascular hypertension. Nutrients 2020; 12; 3321.
Hou WC, Lin RD, Lee TH, Huang YH, Hsu FL, Lee MH. The phenolic constituents and free radical scavenging activities of Gynura formosana Kiamnra. J Sci Food Agri. 2005; 85; 615-21.
Jin X, Wang J, Xia Z, Miao X, Yin Y, Zhang Y. Gynura divaricata extract inhibits hepatocellular carcinoma growth via suppression of Wnt/β-catenin pathway. J Ethnopharmacol. 2018; 226; 141-49.
Kalsoom A, Altaf A, Sattar H, Maqbool T, Sajjad M, Jilani MJ, Shabbir G, Aftab S. Gene expression and anticancer evaluation of Kigelia africana (Lam.) Benth. extracts using MDA-MB-231 and MCF-7 cell lines. PloS one. 2024; 19; e0303134.
Kang H, Kwak Y, Koppula S. Protective effect of purple sweet potato (Ipomoea batatas Linn, Convolvulaceae) on neuro-inflammatory responses in lipopolysaccharide-stimulated microglial cells. Trop J Pharm Res. 2014; 13; 1257-63.
Kang R, Zeh HJ, Lotze MT, Tang D. The beclin 1 network regulates autophagy and apoptosis. Cell Death Differ. 2011; 18; 571-80.
Kantawong F, Saisuwan C, Soeratanapant P, Wanachantararak P, Nan J, Wu J, Chang YT. Gynura divaricata water extract presented the possibility to enhance neuronal regeneration. Evid Based Complement Alternat Med. 2021; 8818618.
Kim HJ. Xu L, Chang KC, Shin SC, Chung J I, Kang D, Kim SH, Hur JA, Choi TH, Kim S, Choi J. Anti-inflammatory effects of anthocyanins from black soybean seed coat on the keratinocytes and ischemia-reperfusion injury in rat skin flaps. Microsurgery 2012; 32; 1-8.
Kosugi H, Kikugawa K. Thiobarbituric acid reaction of aldehydes and oxidized lipids in glacial acetic acid. Lipids 1985; 20; 915-20.
Kumaran A, Karunakaran RJ. Antioxidant and free radical scavenging activity of an aqueous extract of Coleus aromaticus. Food Chem. 2006; 97; 109-14.
Kumar P, Nagarajan A, Uchil PD. Analysis of cell viability by the MTT assay. Cold Spring Harb Protocol. 2018, p 6.
Lee SL, Chin TY, Tu SC, Wang YJ, Hsu YT, Kao MC, Wu YC. Purple sweet potato leaf extract induces apoptosis and reduces inflammatory adipokine expression in 3T3-L1 differentiated adipocytes. Evid Based Complement Alternat Med. 2015; 126302.
Li J, Feng J, Wei H. The aqueous extract of Gynura divaricata (L.) DC. improves glucose and lipid metabolism and ameliorates type 2 diabetes mellitus. Evid Based Complement Alternat Med. 2018; 8686297.
Li J, Ma J, Wang KS, Mi C, Wang Z, Piao LX, Xu GH, Li X, Lee JJ, Jin X. Baicalein inhibits TNF-α-induced NF-κB activation and expression of NF-κB-regulated target gene products. Oncol Rep. 2016; 36; 2771-76.
Lin KH, Hsu CY, Huang YP, Lai JY, Hsieh WB, Huang MY, Yang CM, Chao PY. Chlorophyll-related compounds inhibit cell adhesion and inflammation in human aortic cells. J Med Food. 2013; 16; 886-98.
Lin KH, Lu CP, Chao JW, Yu YP. Antioxidant properties and anti-inflammatory effects of the hydroethanolic extracts of two varieties of bayberry fruit (Myrica rubra Sieb et Zucc.) prepared by stirring and ultrasonic methods. Not Bot Horti Agrobo. 2019; 47; 634-42.
Lin KH, Pan SF, Chen CC, Li WS, Chiang CM. Expression of human papillomavirus type 52 L1 capsid gene in Oryza sativa involved in cytoprotective activities. Not Bot Horti Agrobo. 2020; 48; 40-52.
Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2ΔΔCT method. Methods 2002; 25; 402-08.
Ma JF, Wei PF, Guo C, Shi YP, Lv Y, Qiu LX, Wen LP. The ethyl acetate extract of Gynura formosana Kitam. leaves inhibited cervical cancer cell proliferation via induction of autophagy. Biomed Res Int. 2018; 4780612.
Maqbool T, Faheem H, Sadia N, Inamullah I, Muhammad ABA, Muhammad R, Rabia R, Muhammad A, Awais A, Tahir M. Pulegone induced apoptotic and anti-proliferative potential in cervical cancer via P53, BAX and BCL2. Sci Rep. 2025; 15; 34217.
Morales M, Munne-Bosch S. Malondialdehyde: facts and artifacts. Plant Physiol. 2019; 180; 1246-50.
Nadechanok J, Boonsom L, Saisunee L, John K, Stephen G. The chemical constituents and biological activities of the essential oil and the extracts from leaves of Gynura divaricata (L.) DC. growing in Thailand. J Essent Oil Bear Plant. 2015; 18; 543-55.
Nakano Y, Asada K. Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant Cell Physiol. 1981; 22; 867-80.
Napetschnig J, Wu H. Molecular basis of NF-κB signaling. Ann Rev Biophys. 2013; 42; 443-68.
Nguyen HC, Lin KH, Ho SL, Chiang CM, Yang CM. Enhancing the abiotic stress tolerance of plants: From chemical treatment to biotechnological approaches. Physiol Plant. 2018; 164; 452-66.
Oyaizu M. Antioxidative activities of browning products of glucosamine fractionated by organic solvent and thin layer chromatography. J Jpn Soc Nutr Food Sci. 1988; 35; 771-75.
Pasha A, Kumar K, Heena SK, Emerson IA. Pawar SC. Inhibition of NF‑kB and COX‑2 by andrographolide regu-lates the progression of cervical cancer by promoting PTEN expression and suppressing PI3K/AKT signalling pathway. Sci Rep. 2024; 14; 12020.
Rahman M, Asaeda T, Fukahori K, Imamura F, Nohara A, Matsubayashi M. Hydrogen peroxide measurement can be used to monitor plant oxidative stress rapidly using modified ferrous oxidation xylenol orange and titanium sulfate assay correlation. Int J Plant Biol. 2023; 14; 546-57.
Rifai FNP, Hidayah N, Sakinah FN, Adityani R. Cytotoxic and antiproliferative effects of Gynura divaricata ethanolic extract on HCT-116 colorectal cancer cells in vitro. Int. J. Cell Biomed. Sci. 2024; 3; 217-23.
Ryu R, Kim YJ, Kim KH. Antiproliferative activity of flavonoids from Gynura divaricata in breast cancer cell lines. Pharm Biol. 2017; 55; 2143-49.
Shen Z, Xu J, Wen L, Yin L, Cao X, Pei H, Zhao X. Bioassay-guide preparative separation of hypoglycemic components from Gynura divaricata (L.) DC by conventional and pH-zone refining countercurrent chromatography. Foods 2025; 4; 578.
Sun Y, Gao C. Liu H, Liu X, Yue T. Exploring the mechanism by which aqueous Gynura divaricata inhibits diabetic foot based on network pharmacology, molecular docking and experimental verification. Mol Med. 2023; 29; 11.
Wan C, Yu Y, Zhou S, Liu W, Tian S, Cao S. Antioxidant activity and free radical-scavenging capacity of Gynura divaricata leaf extracts at different temperatures. Pharmacogn Mag. 2011; 7; 40-45.
Xu B. Zhang Y. Bioactive components of Gynura divaricata and its potential use in health, food, and medicine: A mini-review. Afr J Tradit Complement Altern Med. 2017; 14; 113-27.
Xu C, Hu L, Zeng J, Wu A, Deng S, Zhao Z, Geng K. Luo J, Wang L, Zhou X, Huang W, Long Y, Song J, Zheng S, Wu J, Chen Q. Gynura divaricata (L.) DC. promotes diabetic wound healing by activating Nrf2 signaling in diabetic rats. J Ethnopharmacol. 2024; 323; 117638.
Xu W, Lu Z, Wang X, Cheung MH, Chen Y. Gynura divaricata exerts hypoglycemic effects by regulating the PI3K/AKT signaling pathway and fatty acid metabolism signaling pathway. Nutr Diabetes. 2020; 10; 31.
Yao C, Cao X, Fu Z, Tian J, Dong W, Xu J, An K, Zhai L, Yu J. Boschniakia rossica polysaccharide triggers laryngeal carcinoma cell apoptosis by regulating expression of Bcl‑2, caspase‑3, and p53. Med Sci Monit. 2017; 23; 2059-64.
Ye X, Wang L, Yang X, Yang J, Zhou J, Lan C, Kantawong F, Kumsaiyai W, Wu J, Zeng J. Integrated chemical characterization, network pharmacology and transcriptomics to explore the mechanism of sesquiterpenoids isolated from Gynura divaricata (L.) DC. against chronic myelogenous leukemia. Pharmaceuticals 2022; 15; 1435.
Yen CH, Lai CC, Shi TH, Chen M, Yu HC, Liu YP, Chang FR. Gynura divaricata attenuates tumor growth and tumor relapse after cisplatin therapy in HCC xenograft model through suppression of cancer stem cell growth and Wnt/β-catenin signalling. J Ethnopharmacol. 2018; 213; 366-75.
Yoon SS, Moon EY. B-cell adhesion to fibroblast-like synoviocytes is up-regulated by tumor necrosis factor-alpha via expression of human vascular cell adhesion molecule-1 mediated by beta cell-activating factor. Int J Mol Sci. 2021; 22; 7166.
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2026 Lin kuanhung, Shao-Wei Huang, Shwu-Fen Pan, Yi-Hung Li, Chih-Ming Chiang
This work is licensed under a Creative Commons Attribution 4.0 International License.
Authors who publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).
