title: Development of APTES-Decorated HepG2 Cancer Stem Cell Membrane Chromatography for Screening Active Components from Salvia miltiorrhiza.
abstract: Cell membrane chromatography (CMC) is an ideal method for screening potential active components acting on target cell membranes from a complex system, such as herbal medicines. But due to the decay and falling-off of membranes, the CMC column suffers from short life span and low reproducibility. This has greatly limited the application of this model, especially when the cell materials are hard to obtain. To solve this problem, a novel type of (3-aminopropyl)triethoxysilane (APTES)-decorated silica gel was employed. The silica gel was decorated with aldehydes with the help of APTES, which react with the amino groups on cell membranes to form a covalent bond. In this way, cell membranes were immobilized on the surface of silica gel, so it is not easy for membranes to fall off. According to our investigation, the column life of the APTES-decorated group was prolonged to more than 12 days, while the control group showed a sharp decline in column efficiency in the first 3 days. To verify this model, a novel APTES-decorated HepG2 cancer stem cell membrane chromatography (CSCMC) was established and applied in a comprehensive two-dimensional chromatographic system to screen potential active components in Salvia miltiorrhiza. As a result, tanshinone IIA, cryptotanshinone, and dihydrotanshinone I were retained on this model and proved to be effective on HepG2 cancer stem cells by the following cell proliferation and apoptosis assay, with IC50of 10.30 μM, 17.85 μM, and 2.53 μM, respectively. This improvement of CMC can significantly prolong its column life span and broaden the range of its application, which is very suitable for making invaluable or hard-to-obtain cell materials, such as stem cells, for specific drug screening.
authors.affiliation: School of Pharmacy, Second Military Medical University , No. 325 Guohe Road, Shanghai 200433, PR China.
authors.forename: Xuan
authors.initials: X
authors.lastname: Ding
authors.affiliation: School of Pharmacy, Second Military Medical University , No. 325 Guohe Road, Shanghai 200433, PR China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Cao
authors.affiliation: Department of Pharmacy, Shanghai ninth People's Hospital , No. 280 Mohe Road, Shanghai 201999, PR China.
authors.forename: Yongfang
authors.initials: Y
authors.lastname: Yuan
authors.affiliation: School of Pharmacy, Second Military Medical University , No. 325 Guohe Road, Shanghai 200433, PR China.
authors.forename: Zhirong
authors.initials: Z
authors.lastname: Gong
authors.affiliation: School of Pharmacy, Second Military Medical University , No. 325 Guohe Road, Shanghai 200433, PR China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Pharmacy, Eastern Hepatobiliary Surgery Hospital , No. 225 Changhai Road, Shanghai 200438, PR China.
authors.forename: Liang
authors.initials: L
authors.lastname: Zhao
authors.affiliation: Department of Pharmacy, Eastern Hepatobiliary Surgery Hospital , No. 225 Changhai Road, Shanghai 200438, PR China.
authors.forename: Lei
authors.initials: L
authors.lastname: Lv
authors.affiliation: Department of Pharmacy, Eastern Hepatobiliary Surgery Hospital , No. 225 Changhai Road, Shanghai 200438, PR China.
authors.forename: Guoqing
authors.initials: G
authors.lastname: Zhang
authors.affiliation: School of Pharmacy, Second Military Medical University , No. 325 Guohe Road, Shanghai 200433, PR China.
authors.forename: Dongyao
authors.initials: D
authors.lastname: Wang
authors.affiliation: School of Pharmacy, Second Military Medical University , No. 325 Guohe Road, Shanghai 200433, PR China.
authors.forename: Dan
authors.initials: D
authors.lastname: Jia
authors.affiliation: School of Pharmacy, Second Military Medical University , No. 325 Guohe Road, Shanghai 200433, PR China.
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Zhu
authors.affiliation: School of Pharmacy, Second Military Medical University , No. 325 Guohe Road, Shanghai 200433, PR China.
authors.forename: Zhanying
authors.initials: Z
authors.lastname: Hong
authors.affiliation: School of Pharmacy, Second Military Medical University , No. 325 Guohe Road, Shanghai 200433, PR China.
authors.forename: Xiaofei
authors.initials: X
authors.lastname: Chen
authors.affiliation: School of Pharmacy, Second Military Medical University , No. 325 Guohe Road, Shanghai 200433, PR China.
authors.forename: Yifeng
authors.initials: Y
authors.lastname: Chai
mesh_terms: D045784:Abietanes / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001665:Binding Sites; D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D002846:Chromatography, Affinity; D056945:Hep G2 Cells; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D062105:Molecular Docking Simulation; D014411:Neoplastic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D010616:Phenanthrenes / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D010936:Plant Extracts / Q000737:chemistry* / Q000378:metabolism; D011437:Propylamines / Q000737:chemistry*; D027561:Salvia miltiorrhiza / Q000737:chemistry* / Q000378:metabolism; D012821:Silanes / Q000737:chemistry*; D058428:Silica Gel / Q000737:chemistry*; D014183:Transplantation, Heterologous; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000737:chemistry
chemical_list: D045784:Abietanes; D010616:Phenanthrenes; D010936:Plant Extracts; D011437:Propylamines; D012821:Silanes; C021751:tanshinone; C037886:cryptotanshinone; D058428:Silica Gel; D040301:Vascular Endothelial Growth Factor Receptor-2; C477625:amino-propyl-triethoxysilane