title: Chelidonine induces mitotic slippage and apoptotic-like death in SGC-7901 human gastric carcinoma cells.
abstract: The aim of the present study was to investigate the effect of chelidonine on mitotic slippage and apoptotic-like death in SGC-7901 human gastric cancer cells. The MTT assay was performed to detect the antiproliferative effect of chelidonine. Following treatment with chelidonine (10 µmol/l), the ultrastructure changes in SGC-7901, MCF-7 and HepG2 cells were observed by transmission electron microscopy. The effects of chelidonine on G2/M phase arrest and apoptosis of SGC-7901 cells were determined by flow cytometry. Indirect immunofluorescence assay and laser scanning confocal microscopy (LSCM) were used to detect the phosphorylation level of histone H3 (Ser10) and microtubule formation was detected using LSCM following immunofluorescent labeling. Subsequent to treatment with chelidonine (10 µmol/l), expression levels of mitotic slippage-associated proteins, including BUB1 mitotic checkpoint serine/threonine kinase B (BubR1), cyclin-dependent kinase 1 (Cdk1) and cyclin B1, and apoptosis-associated protein, caspase-3 were examined by western blotting at 24, 48 and 72 h. The half maximal inhibitory concentration of chelidonine was 23.13 µmol/l over 48 h and chelidonine induced G2/M phase arrest of cells. The phosphorylation of histone H3 at Ser10 was significantly increased following treatment with chelidonine for 24 h, indicating that chelidonine arrested the SGC-7901 cells in the M phase. Chelidonine inhibited microtubule polymerization, destroyed microtubule structures and induced cell cycle arrest in the M phase. Giant cells were observed with multiple micronuclei of varying sizes, which indicated that following a prolonged arrest in the M phase, the cells underwent mitotic catastrophe. Western blotting demonstrated that the protein expression levels of BubR1, cyclin B1 and Cdk1 decreased significantly between 48 and 72 h. Low expression levels of BubR1 and inactivation of the cyclin B1-Cdk1 complex results in the cells being arrested at mitosis and leads to mitotic slippage. In addition, apoptotic morphological changes in multinucleated cells were observed, the apoptosis rates increased gradually with administration of chelidonine in a time-dependent manner and the protein levels of caspase-3 increased significantly between 24 and 72 h. Thus, chelidonine induces mitotic slippage, and apoptotic-like death occurs in SGC-7901 cells undergoing mitotic catastrophe. Gastric cancer is a common malignancy, and ranks second in overall cancer-associated mortalities worldwide. The present study demonstrated that chelidonine induces M phase arrest and mitotic slippage of SGC-7901 human gastric carcinoma cells via downregulating the expression of BubR1, Cdk1 and cyclin B1 proteins. With the prolongation of chelidonine treatment, the giant cells with multiple micronuclei underwent mitotic slippage and were maintained in the G1 phase and did not survive. A number of multinucleated cells underwent apoptosis via a caspase-dependent signaling pathway. The current study proposes that chelidonine induces mitotic slippage and apoptotic-like death of SGC-7901 cells.
authors.affiliation: School of Pharmacy, Harbin University of Commerce, Harbin, Heilongjiang 150076, P.R. China.
authors.forename: Zhongyuan
authors.initials: Z
authors.lastname: Qu
authors.affiliation: Engineering Research Center of Natural Antineoplastic Drugs, Harbin University of Commerce, Harbin, Heilongjiang 150076, P.R. China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Zou
authors.affiliation: School of Pharmacy, Harbin University of Commerce, Harbin, Heilongjiang 150076, P.R. China.
authors.forename: Xiujuan
authors.initials: X
authors.lastname: Zhang
authors.affiliation: School of Pharmacy, Harbin University of Commerce, Harbin, Heilongjiang 150076, P.R. China.
authors.forename: Jiejing
authors.initials: J
authors.lastname: Sheng
authors.affiliation: Engineering Research Center of Natural Antineoplastic Drugs, Harbin University of Commerce, Harbin, Heilongjiang 150076, P.R. China.
authors.forename: Yumeng
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Engineering Research Center of Natural Antineoplastic Drugs, Harbin University of Commerce, Harbin, Heilongjiang 150076, P.R. China.
authors.forename: Jiaqi
authors.initials: J
authors.lastname: Wang
authors.affiliation: Engineering Research Center of Natural Antineoplastic Drugs, Harbin University of Commerce, Harbin, Heilongjiang 150076, P.R. China.
authors.forename: Chao
authors.initials: C
authors.lastname: Wang
authors.affiliation: Engineering Research Center of Natural Antineoplastic Drugs, Harbin University of Commerce, Harbin, Heilongjiang 150076, P.R. China.
authors.forename: Yubin
authors.initials: Y
authors.lastname: Ji
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D053119:Benzophenanthridines / Q000008:administration & dosage*; D016203:CDC2 Protein Kinase / Q000096:biosynthesis; D002277:Carcinoma / Q000188:drug therapy* / Q000473:pathology / Q000648:ultrastructure; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D045744:Cell Line, Tumor; D056744:Cyclin B1 / Q000096:biosynthesis; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D008938:Mitosis / Q000187:drug effects; D013274:Stomach Neoplasms / Q000188:drug therapy* / Q000473:pathology / Q000648:ultrastructure
chemical_list: D053119:Benzophenanthridines; D056744:Cyclin B1; C062047:chelidonine; D016203:CDC2 Protein Kinase