title: Construction and application of a lung cancer stem cell model: antitumor drug screening and molecular mechanism of the inhibitory effects of sanguinarine.
abstract: Lung cancer is a neoplasm with a 5-year survival rate of less than 15 % and a leading cause of death worldwide, despite recent progress in treatment and diagnostic methods. Lung cancer stem-like cells (CSCs) are pivotal in lung cancer metastasis and drug resistance. This study aimed to develop lung CSCs that stably express stem cell properties through transfection to further screen traditional Chinese herbal compounds. Lung adenocarcinoma stem cells, which include various phenotypic subgroups, are normally characterized by high expression levels of pluripotent stem cell genes, particularly Nanog and OCT4. Plasmids containing Nanog and OCT4 were constructed and transfected into cells, and lung CSCs were identified not only in vitro using RT-PCR, Western blotting, plate cloning, sphere formation, drug resistance, and transwell migration but also in vivo using a nude mouse tumorigenicity assay. Subsequently, sanguinarine, which is derived from the whole leaves of the traditional Chinese medicine celandine, was identified through the high-throughput screening of a small-molecule compound library. Investigation of the molecular mechanisms of the effects of sanguinarine revealed that it significantly inhibited lung CSC proliferation, invasion, and apoptosis, possibly via downregulation of the Wnt/β-catenin signaling pathway. Our results indicate that lung CSCs established by gene transfection may provide a stable and effective method of constructing CSCs to effectively screen potential antitumor drugs. Furthermore, these results suggest that sanguinarine may be a natural antitumor compound that targets lung CSCs, laying a foundation for further clinical study.
authors.affiliation: Department of Oncology, Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Middle 274 Zhijiang Road, Shanghai, 200071, China.|Shanghai University of Traditional Chinese Medicine, 1200 Cai Lun Road, Shanghai, 201203, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Yang
authors.affiliation: Department of Oncology, Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Middle 274 Zhijiang Road, Shanghai, 200071, China.
authors.forename: Zhihong
authors.initials: Z
authors.lastname: Fang
authors.affiliation: Department of Oncology, Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Middle 274 Zhijiang Road, Shanghai, 200071, China.
authors.forename: Jianchun
authors.initials: J
authors.lastname: Wu
authors.affiliation: Department of Oncology, Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Middle 274 Zhijiang Road, Shanghai, 200071, China.
authors.forename: Xiaoling
authors.initials: X
authors.lastname: Yin
authors.affiliation: Department of Oncology, Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Middle 274 Zhijiang Road, Shanghai, 200071, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Fang
authors.affiliation: Department of Oncology, Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Middle 274 Zhijiang Road, Shanghai, 200071, China.
authors.forename: Fanchen
authors.initials: F
authors.lastname: Zhao
authors.affiliation: Shanghai University of Traditional Chinese Medicine, 1200 Cai Lun Road, Shanghai, 201203, China. jusgo105@hotmail.com.
authors.forename: Shiguo
authors.initials: S
authors.lastname: Zhu
authors.affiliation: Department of Oncology, Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Middle 274 Zhijiang Road, Shanghai, 200071, China. 18916767226@163.com.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis; D053119:Benzophenanthridines / Q000494:pharmacology*; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D002465:Cell Movement; D049109:Cell Proliferation; D002945:Cisplatin / Q000494:pharmacology; D019008:Drug Resistance, Neoplasm / Q000187:drug effects; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D006801:Humans; D007124:Immunoenzyme Techniques; D007546:Isoquinolines / Q000494:pharmacology*; D008175:Lung Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D008819:Mice, Nude; D008954:Models, Biological; D009361:Neoplasm Invasiveness; D014411:Neoplastic Stem Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D010936:Plant Extracts / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured; D060449:Wnt Signaling Pathway / Q000187:drug effects*; D014945:Wound Healing; D023041:Xenograft Model Antitumor Assays
chemical_list: D000970:Antineoplastic Agents; D053119:Benzophenanthridines; D014408:Biomarkers, Tumor; D007546:Isoquinolines; D010936:Plant Extracts; D012333:RNA, Messenger; C005705:sanguinarine; D002945:Cisplatin
keywords: Antitumor drug screen; Lung cancer stem cell; Molecular mechanism; Sanguinarine; Wnt/β-catenin signaling