title: Characterization of anti-leukemia components from Indigo naturalis using comprehensive two-dimensional K562/cell membrane chromatography and in silico target identification.
abstract: Traditional Chinese Medicine (TCM) has been developed for thousands of years and has formed an integrated theoretical system based on a large amount of clinical practice. However, essential ingredients in TCM herbs have not been fully identified, and their precise mechanisms and targets are not elucidated. In this study, a new strategy combining comprehensive two-dimensional K562/cell membrane chromatographic system and in silico target identification was established to characterize active components from Indigo naturalis, a famous TCM herb that has been widely used for the treatment of leukemia in China, and their targets. Three active components, indirubin, tryptanthrin and isorhamnetin, were successfully characterized and their anti-leukemia effects were validated by cell viability and cell apoptosis assays. Isorhamnetin, with undefined cancer related targets, was selected for in silico target identification. Proto-oncogene tyrosine-protein kinase (Src) was identified as its membrane target and the dissociation constant (Kd) between Src and isorhamnetin was 3.81 μM. Furthermore, anti-leukemia effects of isorhamnetin were mediated by Src through inducing G2/M cell cycle arrest. The results demonstrated that the integrated strategy could efficiently characterize active components in TCM and their targets, which may bring a new light for a better understanding of the complex mechanism of herbal medicines.
authors.affiliation: School of Biomedical Science, Institute of Molecular Medicine, Huaqiao University, Quanzhou 362021, PR China.|School of Pharmacy, Shanghai Changzheng Hospital, Second Military Medical University, Shanghai 200433, PR China.
authors.forename: Xunxun
authors.initials: X
authors.lastname: Wu
authors.affiliation: School of Pharmacy, Shanghai Changzheng Hospital, Second Military Medical University, Shanghai 200433, PR China.
authors.forename: Xiaofei
authors.initials: X
authors.lastname: Chen
authors.affiliation: School of Pharmacy, Shanghai Changzheng Hospital, Second Military Medical University, Shanghai 200433, PR China.
authors.forename: Jia
authors.initials: J
authors.lastname: Dan
authors.affiliation: School of Pharmacy, Shanghai Changzheng Hospital, Second Military Medical University, Shanghai 200433, PR China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Cao
authors.affiliation: School of Pharmacy, Shanghai Changzheng Hospital, Second Military Medical University, Shanghai 200433, PR China.
authors.forename: Shouhong
authors.initials: S
authors.lastname: Gao
authors.affiliation: School of Biomedical Science, Institute of Molecular Medicine, Huaqiao University, Quanzhou 362021, PR China.|School of Pharmacy, Shanghai Changzheng Hospital, Second Military Medical University, Shanghai 200433, PR China.
authors.forename: Zhiying
authors.initials: Z
authors.lastname: Guo
authors.affiliation: Department of Plant Biology, University of California, Davis, CA 95616, USA.
authors.forename: Philipp
authors.initials: P
authors.lastname: Zerbe
authors.affiliation: School of Pharmacy, Shanghai Changzheng Hospital, Second Military Medical University, Shanghai 200433, PR China.
authors.forename: Yifeng
authors.initials: Y
authors.lastname: Chai
authors.affiliation: School of Biomedical Science, Institute of Molecular Medicine, Huaqiao University, Quanzhou 362021, PR China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Diao
authors.affiliation: School of Pharmacy, Shanghai Changzheng Hospital, Second Military Medical University, Shanghai 200433, PR China.
authors.forename: Lei
authors.initials: L
authors.lastname: Zhang
mesh_terms: D000970:Antineoplastic Agents / Q000302:isolation & purification / Q000494:pharmacology*; D017209:Apoptosis; D002462:Cell Membrane / Q000737:chemistry* / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D002681:China; D002845:Chromatography; D019295:Computational Biology; D006801:Humans; D029883:Indigofera / Q000737:chemistry*; D020014:K562 Cells / Q000737:chemistry / Q000187:drug effects; D018514:Plant Structures / Q000737:chemistry; D000090063:Proto-Oncogene Mas
chemical_list: D000970:Antineoplastic Agents; C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas