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 apopto...

Opposing Effects of Chelidonine on Tyrosine and Serine Phosphorylation of STAT3 in Human Uveal Melanoma Cells

“…It is plausible to assume that the transient effect exerted by chelidonine on the level of pSer-STAT3 in our experiments reflects the same cyclicality, i.e., chelidonine interferes with the machinery leading to activation of Cdk1. This hypothesis is supported by previous data demonstrating that the initial upregulation of cyclin B level, the activator for Cdk1, was followed by a decline upon chelidonine treatment in a gastric cancer cell line [21]. Furthermore, the level of pSer-STAT3 changed in a similar fashion in nocodazole-treated HeLa cells, where Cdk1 was active, whereas the total level of STAT3 did not change [14].…”

<p>Chelidonine enhances the antitumor effect of lenvatinib on hepatocellular carcinoma cells</p>

“…(1) inducing caspase-dependent or caspase-independent cell death; (2) modulating the telomere length; (3) suppressing the NF-κB pathways; (4) inducing mitotic slippage and apoptotic-like death of human cancer cells. [73][74][75][76][77] In this study, we observed the potential application of chelidonine to the treatment of advanced HCC. We established two invivo HCC growth models to examine the in-vivo activity of chelidonine, namely the subcutaneous tumor model and intrahepatic tumor model.…”

“…Moreover, the biological diversity of the secondary metabolites obtained from natural products provides us with new druggable agents, as well as novel options for chemical synthesis and structural modification 67–72. Chelidonine could function as an antitumor agent via multi-mechanisms: (1) inducing caspase-dependent or caspase-independent cell death; (2) modulating the telomere length; (3) suppressing the NF-κB pathways; (4) inducing mitotic slippage and apoptotic-like death of human cancer cells 73–77. In this study, we observed the potential application of chelidonine to the treatment of advanced HCC.…”

Chelidonine Induces Apoptosis via GADD45a-p53 Regulation in Human Pancreatic Cancer Cells

“…[3][4][5] To be specific, chelidonine treatment induced apoptosis in T98G glioma cells, MCF-7 and SK-BR-3 breast adenocarcinoma, HepG2 hepatoma, HeLa cervical cancer, SW620 colon cancer, head and neck squamous cell carcinoma HNSCC, human gastric carcinoma SGC-7901, and leukemia MT-4 cells, through caspase, cell cycle checkpoints and MAP Kinase pathways. 3,[6][7][8][9][10][11][12] In colon cancer Caco-2 and leukemia cell line CEM/ADR 5000, metabolic enzyme regulation by chelidonine reversed doxorubicin resistance. 13 Chelidonine was also reported to trigger autophagy, cellular senescence and blocking telomerase activity.…”