{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T05:11:10Z","timestamp":1790572270208,"version":"4.1.0"},"reference-count":78,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2023,6,28]],"date-time":"2023-06-28T00:00:00Z","timestamp":1687910400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Fundamental Research Grant Scheme (FRGS), Ministry of Higher Education, Malaysia","award":["FRGS\/1\/2020\/SKK0\/UPM\/02\/9"],"award-info":[{"award-number":["FRGS\/1\/2020\/SKK0\/UPM\/02\/9"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>Since lung cancer is the leading cause of cancer-related death worldwide, research is being conducted to discover anticancer agents as its treatment. Eleutherine bulbosa, a Dayak folklore medicine, exhibited anticancer effects against several cancer cells; however, its anticancer potency against lung cancer cells has not been explored yet. This study aims to determine the anticancer potency of E. bulbosa bulbs against lung cancer cells (A549) using 2D and 3D culture models, as well as determine its active compounds using gas chromatography-mass spectrometry (GC-MS) analysis. Three fractions of E. bulbosa bulbs, namely chloroform, n-hexane, and ethyl acetate, were tested for cytotoxicity using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium-bromide (MTT) and CellTiter-Glo. The antiproliferative effects of the most cytotoxic fraction against the 2D culture model were determined by a clonogenic survival assay and propidium iodide\/Hoechst 33342 double staining, whereas the effects against the 3D culture model were determined by microscopy, flow cytometry, and gene expression analysis. The chloroform fraction is the most cytotoxic against A549 cells than other fractions, and it inhibited colony formation and induced apoptosis of A549 cells. The chloroform fraction also inhibited the growth of the A549 spheroid by suppressing the spheroid size, inducing apoptosis, reducing the proportion of CD44 lung cancer stem cells, causing arrest at the S phase of the cell cycle, and suppressing the expression of the SOX2 and MYC genes. Furthermore, the GC-MS analysis detected 20 active compounds in the chloroform fraction, including the major compounds of eleutherine and isoeleutherine. In conclusion, the chloroform fraction of E. bulbosa bulbs exhibit its antiproliferative effect on 2D and 3D culture models of A549 cells, suggesting it could be a lung cancer chemopreventive agent.<\/jats:p>","DOI":"10.3390\/ph16070936","type":"journal-article","created":{"date-parts":[[2023,6,29]],"date-time":"2023-06-29T01:53:57Z","timestamp":1688003637000},"page":"936","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["The Antiproliferative Effect of Chloroform Fraction of Eleutherine bulbosa (Mill.) Urb. on 2D- and 3D-Human Lung Cancer Cells (A549) Model"],"prefix":"10.3390","volume":"16","author":[{"given":"Nur Hannan","family":"Zakaria","sequence":"first","affiliation":[{"name":"UPM-MAKNA Cancer Research Laboratory (CANRES), Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia"},{"name":"Natural Medicine and Product Research Laboratory (NaturMeds), Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Norazalina","family":"Saad","sequence":"additional","affiliation":[{"name":"UPM-MAKNA Cancer Research Laboratory (CANRES), Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Che Azurahanim","family":"Che Abdullah","sequence":"additional","affiliation":[{"name":"UPM-MAKNA Cancer Research Laboratory (CANRES), Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia"},{"name":"Department of Physics, Faculty of Science, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia"},{"name":"Materials Synthesis and Characterization Laboratory (MSCL), Institute of Nanoscience and Nanotechnology (ION2), Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1545-0306","authenticated-orcid":false,"given":"Norhaizan","family":"Mohd. Esa","sequence":"additional","affiliation":[{"name":"Natural Medicine and Product Research Laboratory (NaturMeds), Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia"},{"name":"Department of Nutrition, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,6,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"778","DOI":"10.1002\/ijc.33588","article-title":"Cancer statistics for the year 2020: An overview","volume":"149","author":"Ferlay","year":"2021","journal-title":"Int. J. Cancer"},{"key":"ref_2","unstructured":"(2022, August 15). National Strategic Plan for Cancer Control Programme. 2016\u20132020. Available online: https:\/\/www.iccp-portal.org\/plans\/national-strategic-plan-cancer-control-programme."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"94","DOI":"10.9790\/0853-15100494101","article-title":"Lung cancer: Risk factors, management, and prognosis","volume":"15","author":"Mustafa","year":"2016","journal-title":"IOSR J. Dent. Med. Sci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"45","DOI":"10.5114\/wo.2021.103829","article-title":"Epidemiology of lung cancer","volume":"25","author":"Thandra","year":"2021","journal-title":"Wspolczesna Onkol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1016\/j.bbcan.2015.08.002","article-title":"Lung cancer: Biology and treatment options","volume":"1856","author":"Hassan","year":"2015","journal-title":"Biochim. Biophys. Acta Rev. Cancer"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"9181549","DOI":"10.1155\/2020\/9181549","article-title":"The synergistic anticancer effect of dual drug- (cisplatin\/epigallocatechin gallate) loaded gelatin nanoparticles for lung cancer treatment","volume":"2020","author":"Chen","year":"2020","journal-title":"J. Nanomater."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2396","DOI":"10.1002\/cam4.3660","article-title":"Perspectives and controversies regarding the use of natural products for the treatment of lung cancer","volume":"10","author":"Wen","year":"2021","journal-title":"Cancer Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"303","DOI":"10.3390\/metabo2020303","article-title":"A historical overview of natural products in drug discovery","volume":"2","author":"Dias","year":"2012","journal-title":"Metabolites"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1016\/S2589-3777(19)30018-7","article-title":"Recommended medicinal plants as source of natural products: A review","volume":"1","author":"Santos","year":"2018","journal-title":"Digit. Chin. Med."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"e27163","DOI":"10.1097\/MD.0000000000027163","article-title":"Traditional herbal medicine combined with firstline platinum-based chemotherapy for advanced non-small-cell lung cancer A PRISMA-compliant systematic review and meta-Analysis","volume":"100","author":"Jin","year":"2021","journal-title":"Medicine"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1016\/j.jpha.2018.05.003","article-title":"Primula vulgaris extract induces cell cycle arrest and apoptosis in human cervix cancer cells","volume":"8","author":"Demir","year":"2018","journal-title":"J. Pharm. Anal."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1007\/s11418-010-0396-7","article-title":"Antidermatophyte and antimelanogenesis compound from Eleutherine americana grown in Indonesia","volume":"64","author":"Kusuma","year":"2010","journal-title":"J. Nat. Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1016\/j.proche.2014.12.032","article-title":"Recent studies on phytochemicals and pharmacological effects of Eleutherine americana Merr","volume":"13","author":"Insanu","year":"2014","journal-title":"Procedia Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/j.jacme.2015.05.004","article-title":"Antibacterial activity of Eleutherine bulbosa against multidrug-resistant bacteria","volume":"5","author":"Padhi","year":"2015","journal-title":"J. Acute Med."},{"key":"ref_15","first-page":"286","article-title":"Eleutherine bulbous (Mill.) Urb.: A review study","volume":"10","author":"Couto","year":"2016","journal-title":"J. Med. Plants Res."},{"key":"ref_16","first-page":"273","article-title":"Pharmacognostic and physicochemical evaluation of bulbs of Eleutherine bulbosa (Miller) Urban, a medicinal plant","volume":"4","author":"Rani","year":"2015","journal-title":"J. Pharmacogn. Phytochem."},{"key":"ref_17","first-page":"71","article-title":"Ekstraksi antioksidan bawang Dayak (Eleutherine palmifolia) dengan metode ultrasonic bath (Antioxidant extraction of dayak onion (Eleutherine palmifolia) using ultrasonic bath method)","volume":"5","author":"Yuswi","year":"2018","journal-title":"J. Pangan Agroindustri"},{"key":"ref_18","first-page":"123","article-title":"Anticancer activity of Eleutherine bulbosa (Mill.) Urb. extract on WiDr cell line in vitro","volume":"Volume 9","author":"Lubis","year":"2018","journal-title":"Proceedings of the 2nd Public Health International Conference (PHICo 2017)"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"114770","DOI":"10.1016\/j.jep.2021.114770","article-title":"Induction of apoptosis by Eleutherine bulbosa (mill.) urb. bulb extracted under optimised extraction condition on human retinoblastoma cancer cells (WERI-RB-1)","volume":"284","author":"Kamarudin","year":"2022","journal-title":"J. Ethnopharmacol."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Kamarudin, A.A., Sayuti, N.H., Saad, N., Razak, N.A., and Esa, N.M. (2021). Eleutherine bulbosa (Mill.) urb. bulb: Review of the pharmacological activities and its prospects for application. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22136747"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2000479","DOI":"10.1002\/adfm.202000479","article-title":"Simultaneous culturing of cell monolayers and spheroids on a single microfluidic device for bridging the gap between 2D and 3D cell assays in drug research","volume":"30","author":"Bonabi","year":"2020","journal-title":"Adv. Funct. Mater."},{"key":"ref_22","unstructured":"Ballav, S., Jaywant Deshmukh, A., Siddiqui, S., Aich, J., and Basu, S. (2020). Cell Culture\u2014Advanced Technology and Applications in Medical and Life Sciences, IntechOpen."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"73","DOI":"10.5530\/pj.2021.13.11","article-title":"Standardization of Eleutherine bulbosa Urb. bulbs and total flavonoid content from three locations in Kalimantan, Indonesia","volume":"13","author":"Muthia","year":"2021","journal-title":"Pharmacogn. J."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1186\/s40543-018-0146-0","article-title":"Cytotoxicity evaluation of a Thai herb using tetrazolium (MTT) and sulforhodamine B (SRB) assays","volume":"9","author":"Vajrabhaya","year":"2018","journal-title":"J. Anal. Sci. Technol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"14","DOI":"10.5530\/pj.2020.12.3","article-title":"In vitro cytotoxicity of hibiscus SABDARIFFA Linn extracts on A549 lung cancer cell line","volume":"12","author":"Fithrotunnisa","year":"2020","journal-title":"Pharmacogn. J."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1007\/s11538-020-00746-7","article-title":"Comparison of drug inhibitory effects (IC50) in monolayer and spheroid cultures","volume":"82","author":"Berrouet","year":"2020","journal-title":"Bull. Math. Biol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1016\/j.jep.2009.03.010","article-title":"In vitro screening for anthelmintic and antitumour activity of ethnomedicinal plants from Thailand","volume":"123","author":"Atjanasuppat","year":"2009","journal-title":"J. Ethnopharmacol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Ogbole, O.O., Segun, P.A., and Adeniji, A.J. (2017). In vitro cytotoxic activity of medicinal plants from Nigeria ethnomedicine on Rhabdomyosarcoma cancer cell line and HPLC analysis of active extracts. BMC Complement. Altern. Med., 17.","DOI":"10.1186\/s12906-017-2005-8"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"157","DOI":"10.14499\/indonesianjpharm30iss3pp157","article-title":"Metabolite profiling of Eleutherine palmifolia (L.) Merr. by HPTLC-densitometry and its correlation with anticancer activities and in vitro toxicity","volume":"30","author":"Mutiah","year":"2019","journal-title":"Indones. J. Pharm."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"12855","DOI":"10.1038\/s41598-019-49374-9","article-title":"In vitro and in vivo evaluation of the anticancer and anti-inflammatory activities of 2-Himachelen-7-ol isolated from Cedrus Libani","volume":"9","author":"Elias","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_31","first-page":"3333","article-title":"Effects of damnacanthal and nordamnacanthal on proliferation, apoptosis, and migration of oral squamous cell carcinoma cells","volume":"18","author":"Shaghayegh","year":"2017","journal-title":"Asian Pac. J. Cancer Prev."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1351","DOI":"10.3390\/cancers3011351","article-title":"Cisplatin as an anti-tumor drug: Cellular mechanisms of activity, drug resistance and induced side effects","volume":"3","author":"Florea","year":"2011","journal-title":"Cancers"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Navanesan, S., Abdul Wahab, N., Manickam, S., and Sim, K.S. (2015). Leptospermum flavescens constituent-LF1 causes cell death through the induction of cell cycle arrest and apoptosis in human lung carcinoma cells. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0135995"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1039\/C7QI00299H","article-title":"A new class of platinum(ii) complexes with the phosphine ligand PTA which show potent anticancer activity","volume":"5","author":"Kljun","year":"2018","journal-title":"Inorg. Chem. Front."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Mrid, R.B., Bouchmaa, N., Bouargalne, Y., Ramdan, B., Karrouchi, K., Kabach, I., El Karbane, M., Idir, A., Zyad, A., and Nhiri, M. (2019). Phytochemical characterization, antioxidant and in vitro cytotoxic activity evaluation of juniperus oxycedrus subsp. oxycedrus needles and berries. Molecules, 24.","DOI":"10.3390\/molecules24030502"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"472","DOI":"10.1002\/cyto.a.22273","article-title":"Automatic cell cloning assay for determining the clonogenic capacity of cancer and cancer stem-like cells","volume":"83A","author":"Fedr","year":"2013","journal-title":"Cytom. Part A"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/B978-0-12-385928-0.00003-1","article-title":"Targeting the unfolded protein response in cancer therapy","volume":"491","author":"Backer","year":"2011","journal-title":"Methods Enzymol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Franco, M.S., Roque, M.C., and Oliveira, M.C. (2019). Short and long-term effects of the exposure of breast cancer cell lines to different ratios of free or co-encapsulated liposomal paclitaxel and doxorubicin. Pharmaceutics, 11.","DOI":"10.3390\/pharmaceutics11040178"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"24387","DOI":"10.1039\/C7RA02478A","article-title":"A novel \u03b2-diiminato manganeseiii complex as the promising anticancer agent induces G0\/g1 cell cycle arrest and triggers apoptosis via mitochondrial-dependent pathways in MCF-7 and MDA-MB-231 human breast cancer cells","volume":"7","author":"Farghadani","year":"2017","journal-title":"RSC Adv."},{"key":"ref_40","first-page":"472","article-title":"Antitumor effect of para-toluenesulfonamide against lung cancer xenograft in a mouse model","volume":"5","author":"Gao","year":"2013","journal-title":"J. Thorac. Dis."},{"key":"ref_41","first-page":"1","article-title":"Differential nuclear staining assay for high-throughput screening to identify cytotoxic compounds","volume":"1","author":"Lema","year":"2011","journal-title":"Curr. Cell. Biochem."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"854","DOI":"10.15343\/0104-7809.20194304854869","article-title":"Effect of cortisol on K562 leukemia cells","volume":"43","year":"2019","journal-title":"Mundo Sa\u00fade"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"012031","DOI":"10.1088\/1755-1315\/391\/1\/012031","article-title":"Apoptotic and necrotic lymphocytes after treatment of stem bark extract of plumeria rubra l in vitro","volume":"391","author":"Kuswanti","year":"2019","journal-title":"IOP Conf. Ser. Earth Environ. Sci."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1038\/sj.cdd.4400937","article-title":"Maintenance of ATP favours apoptosis over necrosis triggered by benzamide riboside","volume":"9","author":"Grusch","year":"2002","journal-title":"Cell Death Differ."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Sato, A., Hiramoto, A., Kim, H.S., and Wataya, Y. (2020). Anticancer strategy targeting cell death regulators: Switching the mechanism of anticancer floxuridine-induced cell death from necrosis to apoptosis. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21165876"},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Saleh, K.A., Aldulmani, S.A., Awwad, N.S., Ibrahium, H.A., Asiri, T.H., and Hamdy, M.S. (2019). Utilization of lithium incorporated mesoporous silica for preventing necrosis and increase apoptosis in different cancer cells. BMC Chem., 13.","DOI":"10.1186\/s13065-019-0535-5"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"012022","DOI":"10.1088\/1742-6596\/1277\/1\/012022","article-title":"Antioxidant and anticancer activity of Eleutherine bulbosa (Mill.) Urb on leukemia cells L1210","volume":"1277","author":"Lestari","year":"2019","journal-title":"J. Phys. Conf. Ser."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"86","DOI":"10.22146\/ijp.1077","article-title":"Bioactivity screening of endophytic fungus eutypa linearis isolated from coleus amboinicus (Lour.)","volume":"32","author":"Gemantari","year":"2021","journal-title":"Indones. J. Pharm."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"112064","DOI":"10.1016\/j.indcrop.2019.112064","article-title":"Heat assisted extraction of phenolic compounds from Eleutherine bulbosa (Mill.) bulb and its bioactive profiles using response surface methodology","volume":"144","author":"Kamarudin","year":"2020","journal-title":"Ind. Crops Prod."},{"key":"ref_50","unstructured":"Malheiros, L.C., de Mello, J.C., and Barbosa, W.L. (2015). Phytochemicals\u2014Isolation Characterisation and Role in Human Health, IntechOpen."},{"key":"ref_51","unstructured":"Syamsul, E.S., Apriliana, A., Supomo Saleh, C. (2018, January 27). Thrombocyte Counts in Mice After the Administration of Chloroform Fraction of Eleutherine palmifolia L (Merr). Proceedings of the International Conference on Pharmaceutical Research and Practice 2018, Universitas Islam Indonesia, Yogyakarta, Indonesia."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1007\/s00005-016-0424-8","article-title":"A quinone-containing compound enhances camptothecin-induced apoptosis of lung cancer through modulating endogenous ROS and Erk Signaling","volume":"65","author":"Chou","year":"2017","journal-title":"Arch. Immunol. Ther. Exp."},{"key":"ref_53","doi-asserted-by":"crossref","unstructured":"Zanoaga, O., Braicu, C., Jurj, A., Rusu, A., Buiga, R., and Berindan-Neagoe, I. (2019). Progress in research on the role of flavonoids in lung cancer. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20174291"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"834","DOI":"10.1007\/s00044-021-02701-0","article-title":"In vitro and in vivo biological activities of azulene derivatives with potential applications in medicine","volume":"30","author":"Bakun","year":"2021","journal-title":"Med. Chem. Res."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"994155","DOI":"10.3389\/fonc.2022.994155","article-title":"A spotlight on alkaloid nanoformulations for the treatment of lung cancer","volume":"12","author":"Sindhoor","year":"2022","journal-title":"Front. Oncol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"e3034","DOI":"10.1002\/btpr.3034","article-title":"Establishment and characterization of an in vitro 3D ovarian cancer model for drug screening assays","volume":"36","author":"Tofani","year":"2020","journal-title":"Biotechnol. Prog."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"4371","DOI":"10.3892\/ol.2016.5221","article-title":"Evaluation by quantitative image analysis of anticancer drug activity on multicellular spheroids grown in 3D matrices","volume":"12","author":"Gomes","year":"2016","journal-title":"Oncol. Lett."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"13987","DOI":"10.1038\/srep13987","article-title":"A spheroid toxicity assay using magnetic 3D bioprinting and real-time mobile device-based imaging","volume":"5","author":"Tseng","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"293","DOI":"10.3389\/fonc.2017.00293","article-title":"High-content monitoring of drug effects in a 3D spheroid model","volume":"7","author":"Mittler","year":"2017","journal-title":"Front. Oncol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"2155","DOI":"10.2147\/DDDT.S108004","article-title":"Real-time monitoring of cisplatin cytotoxicity on three-dimensional spheroid tumor cells. Drug Design","volume":"10","author":"Baek","year":"2016","journal-title":"Dev. Ther."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"404","DOI":"10.1016\/j.tranon.2018.11.009","article-title":"Cyclical treatment of colorectal tumor spheroids induces resistance to MEK inhibitors","volume":"12","author":"Thakuri","year":"2019","journal-title":"Transl. Oncol."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"339344","DOI":"10.1016\/j.aca.2021.339344","article-title":"A 3D printed size-tunable flow-focusing droplet microdevice to produce cell-laden hydrogel microspheres","volume":"1192","author":"Nguyen","year":"2022","journal-title":"Anal. Chim. Acta"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"3697","DOI":"10.15419\/bmrat.v7i4.597","article-title":"Method for in vitro production of cartilage microtissues from scaffold-free spheroids composed of human adipose-derived Stem Cells","volume":"7","author":"Tu","year":"2020","journal-title":"Biomed. Res. Ther."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"45745","DOI":"10.18632\/oncotarget.9935","article-title":"The relevance of using 3D cell cultures, in addition to 2D monolayer cultures, when evaluating breast cancer drug sensitivity and resistance","volume":"7","author":"Breslin","year":"2016","journal-title":"Oncotarget"},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"5972152","DOI":"10.1155\/2019\/5972152","article-title":"Effects of microvesicles on cell apoptosis under hypoxia","volume":"2019","author":"Guo","year":"2019","journal-title":"Oxidative Med. Cell. Longev."},{"key":"ref_66","first-page":"229","article-title":"Tumor spheroids and organoids as preclinical model systems","volume":"33","author":"Baniahmad","year":"2021","journal-title":"Med. Genet."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"589601","DOI":"10.3389\/fonc.2020.589601","article-title":"CD44 in ovarian cancer progression and therapy resistance\u2014A critical role for STAT3","volume":"10","author":"Martincuks","year":"2020","journal-title":"Front. Oncol."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"18","DOI":"10.3389\/fcell.2017.00018","article-title":"CD44: A multifunctional cell surface adhesion receptor is a regulator of progression and metastasis of cancer cells","volume":"5","author":"Senbanjo","year":"2017","journal-title":"Front. Cell Dev. Biol."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1186\/s40164-020-00192-0","article-title":"CD44 as a tumor biomarker and therapeutic target","volume":"9","author":"Xu","year":"2020","journal-title":"Exp. Hematol. Oncol."},{"key":"ref_70","unstructured":"Porres, J.M., Wu, M., and Cheng, W.H. (2013). Bioactive Food as Dietary Interventions for the Aging Population, Elsevier."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"6197","DOI":"10.2147\/OTT.S175810","article-title":"SOX2, a stemness gene, induces progression of NSCLC A549 cells toward anchorage-independent growth and chemoresistance to vinblastine","volume":"11","author":"Choe","year":"2018","journal-title":"OncoTargets Ther."},{"key":"ref_72","first-page":"339","article-title":"STAT3\/Oct-4\/c-Myc Signal Circuit for regulating stemness-mediated doxorubicin resistance of triple-negative breast cancer cells and inhibitory effects of WP1066","volume":"53","author":"Cheng","year":"2018","journal-title":"Int. J. Oncol."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"534","DOI":"10.1038\/cdd.2011.137","article-title":"SOX2 has a crucial role in the lineage determination and proliferation of mesenchymal stem cells through dickkopf-1 and c-myc","volume":"19","author":"Park","year":"2012","journal-title":"Cell Death Differ."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1093\/glycob\/cwv088","article-title":"Galectin-3 modulates the EGFR signalling-mediated regulation of SOX2 expression via c-myc in lung cancer","volume":"26","author":"Kuo","year":"2016","journal-title":"Glycobiology"},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"365","DOI":"10.3746\/pnf.2017.22.4.365","article-title":"Analysis of antioxidant and anti-inflammatory activities of solvent fractions from Rhynchosia nulubilis cultivated with Ganoderma lucidum mycelium","volume":"22","author":"Park","year":"2017","journal-title":"Prev. Nutr. Food Sci."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"5864","DOI":"10.3390\/md12125864","article-title":"Polyoxygenated steroids from the octocoral Leptogorgia Punicea and in vitro evaluation of their cytotoxic activity","volume":"12","author":"Moritz","year":"2014","journal-title":"Mar. Drugs"},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"S275","DOI":"10.4103\/0973-1296.166069","article-title":"Xanthohumol induces apoptosis and S phase cell cycle arrest in A549 non-small cell lung cancer cells","volume":"11","author":"Yong","year":"2015","journal-title":"Pharmacogn. Mag."},{"key":"ref_78","first-page":"1729","article-title":"GC-MS analysis of ethyl acetate extract of Eleutherine bulbosa (Urban) Miller (iridaceae)","volume":"7","author":"Rani","year":"2016","journal-title":"Int. J. Pharm. Sci. Res."}],"container-title":["Pharmaceuticals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8247\/16\/7\/936\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T20:02:51Z","timestamp":1760126571000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8247\/16\/7\/936"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,6,28]]},"references-count":78,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2023,7]]}},"alternative-id":["ph16070936"],"URL":"https:\/\/doi.org\/10.3390\/ph16070936","relation":{},"ISSN":["1424-8247"],"issn-type":[{"value":"1424-8247","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,6,28]]}}}