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This study evaluates how liensinine suppresses cancer cell growth by combining functional assays with transcriptomic profiling. Liensinine’s effects were assessed via cell cycle, colony formation, and apoptosis assays, while protein expression changes and signaling pathway enrichment were derived from in vitro and RNA sequencing experiments, with gene transcription validated by RT-PCR. Results show activation of RHOB-mediated MAPK inhibition and NF-κB pathway activation with downstream inflammatory factor release. Overall, liensinine triggers cell death and restrains hypopharyngeal cancer proliferation through multiple pathways, supporting its potential as a chemotherapeutic candidate.",{"@graph":69,"@context":123},[70,84,106],{"@type":71,"itemListElement":72},"BreadcrumbList",[73,77,79,82],{"item":74,"name":75,"@type":76,"position":8},"https://docshare.wps.com","Home","ListItem",{"item":78,"name":9,"@type":76,"position":14},"https://docshare.wps.com/document/",{"item":80,"name":35,"@type":76,"position":81},"https://docshare.wps.com/document/healthcare/",3,{"item":83,"name":65,"@type":76,"position":19},"https://docshare.wps.com/document/rna-sequencingbased-evaluation-of-the-mechanisms-underlying-the-liensinine-mediated-inhibition-of-hypopharyngeal-cancer-proliferation/356368/",{"url":83,"name":65,"@type":85,"image":86,"author":91,"headline":65,"publisher":94,"fileFormat":97,"inLanguage":63,"description":67,"dateModified":98,"datePublished":99,"encodingFormat":97,"isAccessibleForFree":100,"interactionStatistic":101},"DigitalDocument",{"url":87,"@type":88,"width":89,"height":90},"https://docshare.wps.com/thumbnails/rna-sequencingbased-evaluation-of-the-mechanisms-underlying-the-liensinine-mediated-inhibition-of-hypopharyngeal-cancer-proliferation/356368.png","ImageObject",300,407,{"name":92,"@type":93},"Liam","Person",{"url":74,"name":95,"@type":96},"DocShare","Organization","application/pdf","2026-10-27","2026-09-23",true,{"@type":102,"interactionType":103,"userInteractionCount":105},"InteractionCounter",{"@type":104},"ViewAction",14,{"@type":107,"mainEntity":108},"FAQPage",[109,115,119],{"name":110,"@type":111,"acceptedAnswer":112},"What is the main research goal of this study?","Question",{"text":113,"@type":114},"To determine the mechanisms by which liensinine inhibits hypopharyngeal cancer proliferation, which were previously unknown.","Answer",{"name":116,"@type":111,"acceptedAnswer":117},"How were the inhibitory effects of liensinine on tumor cells measured?",{"text":118,"@type":114},"Through cell cycle, colony formation, and apoptosis assays, alongside protein expression analysis and signaling pathway enrichment using in vitro and RNA sequencing experiments.",{"name":120,"@type":111,"acceptedAnswer":121},"Which signaling pathways are implicated in liensinine’s antitumor action?",{"text":122,"@type":114},"Liensinine activates RHOB to inhibit the MAPK pathway and activates NF-κB signaling, leading to increased downstream inflammatory factors.","https://schema.org",{"og:url":83,"og:type":125,"og:title":65,"og:site_name":95,"og:description":67},"article",{"robots":127,"canonical":83},"index,follow",{"doc_id":129,"site_id":62},356368,1790192951,{"code":4,"msg":5,"data":132},{"doc_id":129,"user_id":133,"nickname":92,"user_avatar":134,"doc_module":4,"category_id":34,"category_name":35,"doc_title":65,"doc_description":67,"doc_content":135,"file_id":136,"file_url":137,"file_type":138,"file_size":139,"view_count":105,"is_deleted":4,"is_public":8,"is_downloadable":8,"audit_status":8,"page_count":140,"language":141,"language_code":63,"site_id":62,"html_lang":63,"table_of_contents":142,"faqs":143,"seo_title":144,"seo_description":67,"update_tm":145,"read_time":36},8796095461564,"https://ap-avatar.wpscdn.com/davatar_155a257f0dc6eb9ab79c44ca47cae57d","RNA sequencing–based evaluation of the mechanisms underlying the liensinine-mediated inhibition of hypopharyngeal cancer proliferation  \nChen Qin, Hong Pan, Dan Zhang, Yinzhe Gaiand Mang Xiao   \nJournal of International Medical Research 2026, Vol. 54(2) 1–16! The Author(s) 2026 Article reuse guidelines:  \n[sagepub.com/journals-permissions](sagepub.com/journals-permissions)[ ](sagepub.com/journals-permissions)[DOI: 10.1177/03000605261419604](DOI: 10.1177/03000605261419604)[ ](DOI: 10.1177/03000605261419604)[journals.sagepub.com/home/imr](journals.sagepub.com/home/imr)  \nAbstract  \nBackground: Hypopharyngeal cancer is increasingly emerging as a disease that threatens global health, with poor prognosis and survival rates. However, clinical strategies and effective therapies remain limited.  \nMethods: The inhibitory effect of liensinine on tumor cells was detected through cell cycle, colony formation, and apoptosis assays. Changes in the expression levels of relevant proteins were detected and enrichment analysis of signaling pathways was performed through in vitro and RNA sequencing experiments. The transcription levels of relevant genes were further verified using reverse transcription polymerase chain reaction.  \nResults: We previously discovered that the natural compound, liensinine, is effective in treating hypopharyngeal cancer. In this study, we found through in vitro and RNA sequencing experiments that liensinine can activate the Ras homolog family member B protein, thereby inhibiting the mitogen-activated protein kinase signaling pathway. Additionally, liensinine activates the nuclear factor kappa B signaling pathway and releases downstream inflammatory factors, effectively exerting its antitumor effects.  \nConclusion: Liensinine induces cell death and inhibits hypopharyngeal cancer cell growth through multiple pathways, indicating that it is a potential chemotherapeutic agent for the treatment of hypopharyngeal cancer.  \nDepartment of Otolaryngology Head and Neck Surgery, Sir Run Run Shaw Hospital, College of Medicine, Zhejiang University, China  \nCorresponding author:  \nMang Xiao, Department of Otolaryngology Head and Neck Surgery, Sir Run Run Shaw Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang, China.  \nEmail: [joelxm@zju.edu.cn](joelxm@zju.edu.cn)  \nCreative Commons Non Commercial CC BY-NC: This article is distributed under the terms of the Creative  \nCommons Attribution-NonCommercial 4.0 License ([https://creativecommons.org/licenses/by-nc/4.0/](https://creativecommons.org/licenses/by-nc/4.0/)) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access pages ([https://us.sagepub.com/en-us/nam/open-access-at-sage](https://us.sagepub.com/en-us/nam/open-access-at-sage)).  \nKeywords  \nHypopharyngeal cancer, liensinine, mitogen-activated protein kinase, nuclear factor kappa B, Ras homolog family member B  \nDate received: 12 August 2025; accepted: 12 January 2026  \nIntroduction  \nHypopharyngeal carcinoma, a subtype of head and neck squamous cell carcinoma (HNSCC), is characterized by its aggressive behavior and poor prognosis. 1,2 Arising from the epithelial lining of the hypopharynx, this malignancy often presents at an advanced stage due to its anatomical location, which complicates early detection.3 The advent of RNA sequencing (RNAseq) has enabled a deeper understanding of the transcriptomic landscape of hypopharyngeal carcinoma, facilitating the identification of novel biomarkers and therapeutic vulnerabilities.4  \nLiensinine is a bisbenzylisoquinoline alkaloid derived from the traditional Chinese medicinal plant Nelumbo nucifera Gaertn.5 It has garnered considerable attention owing to its diverse pharmacological activities, particularly in the context of cancer therapy.6,7 The compound exhibits potent antiproliferative effects across various malignancies, suggesting a p","cbCaikNd5nnnP0tg","https://ap.wps.com/l/cbCaikNd5nnnP0tg","pdf",1599781,16,"English","# Abstract\n## Background\n## Methods\n## Results\n## Conclusion\n# Introduction\n# Materials and methods\n## Cell culture and methods","[{\"question\":\"What is the main research goal of this study?\",\"answer\":\"To determine the mechanisms by which liensinine inhibits hypopharyngeal cancer proliferation, which were previously unknown.\"},{\"question\":\"How were the inhibitory effects of liensinine on tumor cells measured?\",\"answer\":\"Through cell cycle, colony formation, and apoptosis assays, alongside protein expression analysis and signaling pathway enrichment using in vitro and RNA sequencing experiments.\"},{\"question\":\"Which signaling pathways are implicated in liensinine’s antitumor action?\",\"answer\":\"Liensinine activates RHOB to inhibit the MAPK pathway and activates NF-κB signaling, leading to increased downstream inflammatory factors.\"}]","RNA sequencing–based evaluation of the mechanisms underlying the liensinine-mediated inhibition of hypopharyngeal cancer proliferation | PDF",1790125044]