title: Ophiopogonin B induces apoptosis, mitotic catastrophe and autophagy in A549 cells.
abstract: Ophiopogonin B (OP-B), a saponin compound isolated from Radix Ophiopogon japonicus, was verified to inhibit cell proliferation in numerous non-small cell lung cancer (NSCLC) cells in our previous study. However, the precise mechanisms of action have remained unclear. In the present study, we mainly investigated the effects of OP-B on adenocarcinoma A549 cells to further elaborate the underlying mechanisms of OP-B in different NSCLC cell lines. Detection by high content screening (HCS) and TUNEL assay verified that OP-B induced apoptosis in this cell line, while detection of Caspase-3, Bcl-2 and Bax showed that OP-B induced cell death was caspase and mitochondrial independent. Further experiments showed that OP-B induced cell cycle arrest in the S and G2/M phases by inhibiting the expression of Myt1 and phosphorylation of Histone H3 (Ser10), which resulted in mitotic catastrophe in the cells. Transmission electron microscopy (TEM) observation of cell micro-morphology combined with detection of Atgs by western blot analysis showed that OP-B induced autophagy in this cell line. Autophagy inhibition by the lysosome inhibitor CQ or Beclin1-siRNA knockdown both attenuated cell viability, demonstrated that autophagy also being the vital reason resulted in cell death. More importantly, the xenograft model using A549 cells provided further evidence of the inhibition of OP-B on tumor proliferation. Immunohistochemistry detection of LC3 and Tunel assay both verified that high dose of OP-B (75 mg/kg) induced autophagy and apoptosis in vivo, and western blot detection of p-Histone H3 (Ser10), Survivin and XIAP further indicated the molecular mechanism of OP-B in vivo. As our findings revealed, multiple types of cell death overlapped in OP-B treated A549 cells, it displayed multitarget characteristics of the compounds extracted from the Chinese herbal, which may be used as candidate anticancer medicine in clinic.
authors.affiliation: The Pre-Clinical Medicine College, Nanjing University of Chinese Medicine, Nanjing 210023, P.R. China.
authors.forename: Meijuan
authors.initials: M
authors.lastname: Chen
authors.affiliation: The Pre-Clinical Medicine College, Nanjing University of Chinese Medicine, Nanjing 210023, P.R. China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Guo
authors.affiliation: The Pre-Clinical Medicine College, Nanjing University of Chinese Medicine, Nanjing 210023, P.R. China.
authors.forename: Ruolin
authors.initials: R
authors.lastname: Zhao
authors.affiliation: Jiangsu Collaborative Innovation Center of Traditional Chinese Medicine (TCM) Prevention and Treatment of Tumor, Nanjing University of Chinese Medicine, Nanjing 210023, P.R. China.
authors.forename: Xiaoxia
authors.initials: X
authors.lastname: Wang
authors.affiliation: The Pre-Clinical Medicine College, Nanjing University of Chinese Medicine, Nanjing 210023, P.R. China.
authors.forename: Miao
authors.initials: M
authors.lastname: Jiang
authors.affiliation: Department of Pharmacology, Emory University School of Medicine, Atlanta, GA 30322, USA.
authors.forename: Haian
authors.initials: H
authors.lastname: Fu
authors.affiliation: Jiangsu Collaborative Innovation Center of Traditional Chinese Medicine (TCM) Prevention and Treatment of Tumor, Nanjing University of Chinese Medicine, Nanjing 210023, P.R. China.
authors.forename: Xu
authors.initials: X
authors.lastname: Zhang
mesh_terms: D000072283:A549 Cells; D017209:Apoptosis / Q000187:drug effects*; D001343:Autophagy / Q000187:drug effects*; D002289:Carcinoma, Non-Small-Cell Lung / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D059447:Cell Cycle Checkpoints; D049109:Cell Proliferation / Q000187:drug effects; D004365:Drugs, Chinese Herbal / Q000008:administration & dosage; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008938:Mitosis / Q000187:drug effects*; D034741:RNA, Small Interfering; D012503:Saponins / Q000008:administration & dosage*; D013150:Spirostans / Q000008:administration & dosage*; D023041:Xenograft Model Antitumor Assays
chemical_list: D004365:Drugs, Chinese Herbal; D034741:RNA, Small Interfering; D012503:Saponins; D013150:Spirostans; C046997:ophiopogonin B