{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,23]],"date-time":"2026-08-23T14:50:53Z","timestamp":1787496653179,"version":"build-2736575974"},"reference-count":49,"publisher":"Public Library of Science (PLoS)","issue":"10","license":[{"start":{"date-parts":[[2024,10,24]],"date-time":"2024-10-24T00:00:00Z","timestamp":1729728000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002842","name":"Chiang Mai University","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002842","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>\n                    Cyanobacteria and algae serving as promising food supplements have recently garnered attention for their emerging potential in anti-cancer activity. Cholangiocarcinoma (CCA) or bile duct cancer is one of the top-leading cancers affecting people, particularly in Asian continent. With patients exhibiting no or minimal symptoms in the early stages, advanced CCA is often diagnosed, and primary treatments such as surgery may not be suitable. Discovery of natural bioactive compounds for cancer treatments have, thus, attracted attention as one of the effective means to combat CCA or to supplement primary treatments. In this work, ethanolic and polysaccharide extracts of cyanobacteria and algae were tested for their cytotoxicity against 2 CCA cell lines (KKU055 and KKU213A). The ethanolic extracts from\n                    <jats:italic>Leptolyngbya<\/jats:italic>\n                    sp. and\n                    <jats:italic>Chlorella<\/jats:italic>\n                    sp. demonstrated growth inhibition of both CCA cell lines, with IC50 values of 0.658 mg\/mL and 0.687 mg\/mL for KKU055, and 0.656 mg\/mL and 0.450 mg\/mL for KKU213A. In contrast, only the polysaccharide extracts from\n                    <jats:italic>Sargassum<\/jats:italic>\n                    spp. exhibited a remarkable cytotoxic effect, while the polysaccharide extract from\n                    <jats:italic>Spirulina<\/jats:italic>\n                    sp. showed slight effect only at a higher concentration (2 mg\/mL). All tested extracts were further investigated for improving immune cell killing ability and showed that\n                    <jats:italic>Spirulina<\/jats:italic>\n                    sp. polysaccharide extract was able to improve the immune cell killing ability. This extract was then investigated for its effects on the immune cell population, which demonstrated to have positive impact on NK cell population. To further explore the potential use, synergistic effect of\n                    <jats:italic>Spirulina<\/jats:italic>\n                    sp. polysaccharide extract with an already-in-use chemotherapeutic drug, gemcitabine, on immune cell cytotoxicity was investigated. The results showed that the immune cell cytotoxicity was enhanced in the co-treatment compared to the use of each treatment separately. The most apparent difference was observed in KKU055 cells where % living cells were reduced from 78.96% (immune cell alone) to 20.93% when the combined gemcitabine and\n                    <jats:italic>Spirulina<\/jats:italic>\n                    sp. extracts were used.\n                  <\/jats:p>","DOI":"10.1371\/journal.pone.0312414","type":"journal-article","created":{"date-parts":[[2024,10,24]],"date-time":"2024-10-24T13:34:21Z","timestamp":1729776861000},"page":"e0312414","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":5,"title":["Polysaccharide extract of Spirulina sp. increases effector immune-cell killing activities against cholangiocarcinoma"],"prefix":"10.1371","volume":"19","author":[{"given":"Aussara","family":"Panya","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3949-9887","authenticated-orcid":true,"given":"Methi","family":"Wathikthinnakon","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chutamas","family":"Thepmalee","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chutipa","family":"Chiawpanit","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-5545-7948","authenticated-orcid":true,"given":"Suthida","family":"Panwong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yupanun","family":"Wutti-in","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Preeyanat","family":"Vongchan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Phennapha","family":"Klangsinsirikul","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1272-544X","authenticated-orcid":true,"given":"Pachara","family":"Sattayawat","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8878-6607","authenticated-orcid":true,"given":"Jeeraporn","family":"Pekkoh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2024,10,24]]},"reference":[{"key":"pone.0312414.ref001","first-page":"9","article-title":"Anticancer potential of algae-derived metabolites: recent updates and breakthroughs","author":"R Sharma","year":"2023","journal-title":"Futur J Pharm Sci"},{"key":"pone.0312414.ref002","first-page":"18","article-title":"Algae-derived bioactive compounds with anti-lung cancer potential","author":"I Saadaoui","year":"2020","journal-title":"Mar Drugs"},{"key":"pone.0312414.ref003","doi-asserted-by":"crossref","first-page":"1065","DOI":"10.21037\/tcr.2019.06.13","article-title":"The efficacy of dietary Spirulina as an adjunct to chemotherapy to improve immune function and reduce myelosuppression in patients with malignant tumors","volume":"8","author":"Y Ge","year":"2019","journal-title":"Transl Cancer Res"},{"key":"pone.0312414.ref004","article-title":"Epidemiology of cholangiocarcinoma","author":"M Qurashi","year":"2023","journal-title":"Eur J Surg Oncol"},{"key":"pone.0312414.ref005","doi-asserted-by":"crossref","first-page":"618","DOI":"10.1111\/jcmm.16151","article-title":"Therapy for advanced cholangiocarcinoma: Current knowledge and future potential","volume":"25","author":"M Wang","year":"2021","journal-title":"J Cell Mol Med"},{"key":"pone.0312414.ref006","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1016\/j.apjtb.2016.10.011","article-title":"Cytotoxic effect of Spirulina platensis extracts on human acute leukemia Kasumi-1 and chronic myelogenous leukemia K-562 cell lines","volume":"7","author":"FY Flores Hernandez","year":"2017","journal-title":"Asian Pac J Trop Biomed"},{"key":"pone.0312414.ref007","doi-asserted-by":"crossref","first-page":"292","DOI":"10.1016\/j.biopha.2018.06.116","article-title":"Anticancer effect of the water extract of a commercial Spirulina (Arthrospira platensis) product on the human lung cancer A549 cell line","volume":"106","author":"A Czerwonka","year":"2018","journal-title":"Biomed Pharmacother"},{"key":"pone.0312414.ref008","doi-asserted-by":"crossref","first-page":"114306","DOI":"10.1016\/j.biopha.2023.114306","article-title":"Microalga Chlorella sp. extract induced apoptotic cell death of cholangiocarcinoma via AKT\/mTOR signaling pathway","volume":"160","author":"N Sawasdee","year":"2023","journal-title":"Biomed Pharmacother"},{"key":"pone.0312414.ref009","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1007\/s12029-021-00765-0","article-title":"Anticancer Effect of Sargassum oligocystom Hydroalcoholic Extract Against SW742, HT-29, WiDr, and CT-26 Colorectal Cancer Cell Lines and Expression of P53 and APC Genes","volume":"54","author":"IRJ Al-Aadily","year":"2023","journal-title":"J Gastrointest Cancer"},{"key":"pone.0312414.ref010","doi-asserted-by":"crossref","first-page":"102511","DOI":"10.1016\/j.algal.2021.102511","article-title":"Lipophilic extracts of the thermophilic cyanobacterium Leptolyngbya sp. and chlorophyte Graesiella sp. and their potential use as food and anticancer agents","volume":"60","author":"M Gara-Ali","year":"2021","journal-title":"Algal Res"},{"key":"pone.0312414.ref011","doi-asserted-by":"crossref","first-page":"310","DOI":"10.1016\/j.ccell.2020.11.009","article-title":"Immunomodulation by targeted anticancer agents","volume":"39","author":"G Petroni","year":"2021","journal-title":"Cancer Cell"},{"key":"pone.0312414.ref012","doi-asserted-by":"crossref","first-page":"100147","DOI":"10.1016\/j.medidd.2022.100147","article-title":"A review of the pharmacodynamic effect of chemo-herbal drug combinations therapy for cancer treatment","volume":"17","author":"A Okem","year":"2023","journal-title":"Med Drug Discov"},{"key":"pone.0312414.ref013","doi-asserted-by":"crossref","first-page":"1409","DOI":"10.1111\/bph.14816","article-title":"Natural compounds as potential adjuvants to cancer therapy: Preclinical evidence","volume":"177","author":"SR Lin","year":"2020","journal-title":"Br J Pharmacol"},{"key":"pone.0312414.ref014","doi-asserted-by":"crossref","first-page":"106006","DOI":"10.1016\/j.intimp.2019.106006","article-title":"Gemcitabine enhances cytotoxic activity of effector T-lymphocytes against chemo-resistant cholangiocarcinoma cells","volume":"78","author":"N Sawasdee","year":"2020","journal-title":"Int Immunopharmacol"},{"key":"pone.0312414.ref015","doi-asserted-by":"crossref","first-page":"1367","DOI":"10.1016\/j.gendis.2022.02.007","article-title":"Cancer chemotherapy and beyond: Current status, drug candidates, associated risks and progress in targeted therapeutics","volume":"10","author":"U Anand","year":"2023","journal-title":"Genes Dis"},{"key":"pone.0312414.ref016","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1016\/j.biopha.2018.12.069","article-title":"Microalgae in modern cancer therapy: Current knowledge","volume":"111","author":"ME Abd El-Hack","year":"2019","journal-title":"Biomed Pharmacother"},{"key":"pone.0312414.ref017","doi-asserted-by":"crossref","first-page":"1125","DOI":"10.1002\/jobm.202100459","article-title":"Cyanobacteria derived compounds: Emerging drugs for cancer management","volume":"62","author":"A Shahid","year":"2022","journal-title":"J Basic Microbiol"},{"key":"pone.0312414.ref018","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3390\/ph14020157","article-title":"Status and challenges of plant-anticancer compounds in cancer treatment","volume":"14","author":"P Garcia-Oliveira","year":"2021","journal-title":"Pharmaceuticals"},{"key":"pone.0312414.ref019","first-page":"11","article-title":"Naturally Occurring Functional Ingredient from Filamentous Thermophilic Cyanobacterium Leptolyngbya sp. KC45: Phytochemical Characterizations and Their Multiple Bioactivities","author":"K Phinyo","year":"2022","journal-title":"Antioxidants"},{"key":"pone.0312414.ref020","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3390\/app10093058","article-title":"In vitro evaluation of anti-colon cancer potential of crude extracts of fucoidan obtained from sargassum glaucescens pretreated by compressional-puffing","volume":"10","author":"WC Shiao","year":"2020","journal-title":"Appl Sci"},{"key":"pone.0312414.ref021","doi-asserted-by":"crossref","first-page":"1215","DOI":"10.1016\/j.ijbiomac.2016.06.084","article-title":"Cytotoxic effect of fucoidan extracted from Sargassum cinereum on colon cancer cell line HCT-15","volume":"91","author":"SN Somasundaram","year":"2016","journal-title":"Int J Biol Macromol"},{"key":"pone.0312414.ref022","doi-asserted-by":"crossref","first-page":"976","DOI":"10.1016\/j.ijbiomac.2019.05.117","article-title":"Production and characterization of exopolysaccharides from Chlorella zofingiensis and Chlorella vulgaris with anti-colorectal cancer activity","volume":"134","author":"J Zhang","year":"2019","journal-title":"Int J Biol Macromol"},{"key":"pone.0312414.ref023","doi-asserted-by":"crossref","first-page":"7058","DOI":"10.1021\/acs.jafc.2c00176","article-title":"Polysaccharide from Spirulina platensis Evokes Antitumor Activity in Gastric Cancer Cells via Modulation of Galectin-3 and Exhibited Cyto\/DNA Protection: Structure\u2013Function Study","volume":"70","author":"V Uppin","year":"2022","journal-title":"J Agric Food Chem"},{"key":"pone.0312414.ref024","first-page":"275","article-title":"Modulation of the Anti-cancer Activity of Sulfated Polysaccharides, Synthesized in Spirulina platensis, Due to Varying Degree of Sulfation Induced by Nutrient and Physical Stress","volume":"10","author":"VD Mendhulkar","year":"2020","journal-title":"J Biol Act Prod from Nat"},{"key":"pone.0312414.ref025","doi-asserted-by":"crossref","first-page":"106158","DOI":"10.1016\/j.jff.2024.106158","article-title":"Spirulina polysaccharide-based prebiotic foods preparations-a promising approach for modulating gut microbiota and improving health","volume":"116","author":"F Guan","year":"2024","journal-title":"J Funct Foods"},{"key":"pone.0312414.ref026","doi-asserted-by":"crossref","first-page":"3379","DOI":"10.1007\/s10811-016-0881-9","article-title":"Characterization and antitumor activity of protein hydrolysates from Arthrospira platensis (Spirulina platensis) using two-step hydrolysis","volume":"28","author":"Z Wang","year":"2016","journal-title":"J Appl Phycol"},{"key":"pone.0312414.ref027","first-page":"228","article-title":"Composition, isolation, purification and biological activities of Sargassum fusiforme polysaccharides: A review","author":"R Zhang","year":"2020","journal-title":"Carbohydr Polym"},{"key":"pone.0312414.ref028","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1007\/s10811-020-02301-2","article-title":"Effects of extracted polysaccharides from a Chlorella vulgaris biomass on expression of interferon-\u03b3 and interleukin-2 in chicken peripheral blood mononuclear cells","volume":"33","author":"S Mirzaie","year":"2021","journal-title":"J Appl Phycol"},{"key":"pone.0312414.ref029","first-page":"18","article-title":"Immunostimulatory Effects of Polysaccharides from Spirulina platensis In Vivo and Vitro and Their Activation Mechanism on RAW246.7 Macrophages","author":"X Wu","year":"2020","journal-title":"Mar Drugs"},{"key":"pone.0312414.ref030","doi-asserted-by":"crossref","first-page":"1319","DOI":"10.1002\/JLB.MR0718-269R","article-title":"Mechanisms of natural killer cell-mediated cellular cytotoxicity","volume":"105","author":"I Prager","year":"2019","journal-title":"J Leukoc Biol"},{"key":"pone.0312414.ref031","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1111\/j.1365-2567.2009.03123.x","article-title":"Natural killer cell cytotoxicity: How do they pull the trigger?","volume":"128","author":"NJ Topham","year":"2009","journal-title":"Immunology"},{"key":"pone.0312414.ref032","first-page":"17","article-title":"Ascophyllan Induces Activation of Natural Killer Cells in Mice in Vivo and in Vitro","author":"W Zhang","year":"2019","journal-title":"Mar Drugs"},{"key":"pone.0312414.ref033","first-page":"26","article-title":"Cordycepin sensitizes cholangiocarcinoma cells to be killed by natural killer-92 (Nk-92) cells","author":"S Panwong","year":"2021","journal-title":"Molecules"},{"key":"pone.0312414.ref034","first-page":"11","article-title":"Genistein Sensitizes Human Cholangiocarcinoma Cell Lines to Be Susceptible to Natural Killer Cells","author":"C Chiawpanit","year":"2022","journal-title":"Biology (Basel)"},{"key":"pone.0312414.ref035","first-page":"1","article-title":"Synergistic effects of plant derivatives and conventional chemotherapeutic agents: An update on the cancer perspective","volume":"55","author":"R Pezzani","year":"2019","journal-title":"Med"},{"key":"pone.0312414.ref036","first-page":"1","article-title":"Ivermectin and gemcitabine combination treatment induces apoptosis of pancreatic cancer cells via mitochondrial dysfunction","volume":"13","author":"DE Lee","year":"2022","journal-title":"Front Pharmacol"},{"key":"pone.0312414.ref037","first-page":"2018","article-title":"Gemcitabine-based chemotherapy for advanced biliary tract carcinomas","author":"O Abdel-Rahman","year":"2018","journal-title":"Cochrane Database Syst Rev"},{"key":"pone.0312414.ref038","first-page":"239","article-title":"Role of gemcitabine in cancer therapy","volume":"1","author":"RW Brockman","year":"1963","journal-title":"Metab Inhib"},{"key":"pone.0312414.ref039","first-page":"1912","article-title":"Severe acute toxicity following gemcitabine administration: A report of four cases with cytidine deaminase polymorphisms evaluation","volume":"15","author":"B Hryciuk","year":"2018","journal-title":"Oncol Lett"},{"key":"pone.0312414.ref040","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41598-022-09964-6","article-title":"Combination gemcitabine and PD-L1xCD3 bispecific T cell engager (BiTE) enhances T lymphocyte cytotoxicity against cholangiocarcinoma cells","volume":"12","author":"M Wathikthinnakon","year":"2022","journal-title":"Sci Rep"},{"key":"pone.0312414.ref041","doi-asserted-by":"crossref","first-page":"398","DOI":"10.3892\/ijo.2015.3019","article-title":"Establishment and characterization of gemcitabine-resistant human cholangiocarcinoma cell lines with multidrug resistance and enhanced invasiveness","volume":"47","author":"W Wattanawongdon","year":"2015","journal-title":"Int J Oncol"},{"key":"pone.0312414.ref042","first-page":"28","article-title":"Exploration of the Use of Natural Compounds in Combination with Chemotherapy Drugs for Tumor Treatment","author":"J Wu","year":"2023","journal-title":"Molecules"},{"key":"pone.0312414.ref043","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41598-021-94049-z","article-title":"Anticancer effects of the combined Thai noni juice ethanolic extracts and 5-fluorouracil against cholangiocarcinoma cells in vitro and in vivo","volume":"11","author":"J Prompipak","year":"2021","journal-title":"Sci Rep"},{"key":"pone.0312414.ref044","first-page":"1","article-title":"Synergistic effect of the herbal mixture C5E on gemcitabine treatment in PANC-1 cells","volume":"23","author":"PJ Pak","year":"2021","journal-title":"Mol Med Rep"},{"key":"pone.0312414.ref045","doi-asserted-by":"crossref","first-page":"149","DOI":"10.3892\/or.2013.2461","article-title":"Inhibition of pancreatic cancer and potentiation of gemcitabine effects by the extract of Pao Pereira","volume":"30","author":"J Yu","year":"2013","journal-title":"Oncol Rep"},{"key":"pone.0312414.ref046","doi-asserted-by":"crossref","first-page":"1","DOI":"10.21608\/ajfm.2018.18076","article-title":"Pretreatment with the Micro-alga, Spirulina Platensis Ameliorates Cyclophosphamide -Induced Hematological, Liver and Kidney Toxicities in Male Mice","volume":"30","author":"S El-Naggar","year":"2018","journal-title":"Ain Shams J Forensic Med Clin Toxicol"},{"key":"pone.0312414.ref047","doi-asserted-by":"crossref","first-page":"351","DOI":"10.3136\/fstr.14.351","article-title":"A subchronic toxicity study of Spirulina platensis","volume":"14","author":"N Hutadilok-Towatana","year":"2008","journal-title":"Food Sci Technol Res"},{"key":"pone.0312414.ref048","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1007\/BF02934440","article-title":"Effect of spirulina platensis polysaccharide on hematopoietic recovery and related cytokines in mice with transplanted tumor treated by chemotherapy","volume":"8","author":"X Liu","year":"2002","journal-title":"Chinese J Integr Tradit West Med"},{"key":"pone.0312414.ref049","first-page":"13","article-title":"Spirulina (Arthrospira platensis): Antiallergic Agent or Hidden Allergen? A Literature Review","author":"W Gromek","year":"2024","journal-title":"Foods"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0312414","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,10,24]],"date-time":"2024-10-24T13:34:38Z","timestamp":1729776878000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0312414"}},"subtitle":[],"editor":[{"given":"Mohamed","family":"Abdel-Daim","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2024,10,24]]},"references-count":49,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2024,10,24]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0312414","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,10,24]]}}}