The effects of Danthron on the cell cycle and apoptosis of human brain glioblastoma multiforms GBM 8401 cells

• Main Authors: Ying Ying Chuang, 莊穎瑩
• Other Authors: Hai Lung Wang
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/3jrkh6

碩士 === 元培科技大學 === 醫學檢驗生物技術研究所 === 97 === Danthron (1,8-dihydroxyanthraquinone), one of component from Rheum palmatum L. (Polygonaceae), is an effective component of nature Chinese herb medicine and has been shown several biological activities inclusive of with a powerful anti-tumor activity. For the effects on protein function, Danthron is considered as a tyrosine kinase inhibitor. Danthron can not only inhibit tyrosine kinase signal pathway, proliferation and metastasis ability of cancer cells but induce apoptosis of cancer cells, including lung cancer and breast cancer etc. In addition, Danthron also inhibits human cervical epitheliod carcinoma, leukemia, hepatoma, ovarian carcinoma, esophageal carcinoma and glioma cells. The aim of this study is to investigate the mechanisms by danthron for the induction of apoptotic potential on human brain glioblastoma multiforms GBM 8401 cell line. In this study, Danthron -induced cytotoxic effect of GBM 8401 cells were investigated. For determining cell viability, the flow cytometric analysis was used. According to the results of this study, Danthron showed a marked dose- and time-dependent inhibition of GBM 8401 cell viability and revealed G0/G1 phase arrest and sub-G1 occurrence. Danthron also caused a marked cell morphology damage proved by microscopic examination, DAPI staining and comet assay. Examined by flow cytometry, Danthron could affected on the production of a large number of intracellular reactive oxygen species (ROS) and promoted that calcium release from endoplasmic reticulum (ER) and on the decrease of the level of mitochondria membrane potential to damage cancer cells and then cause the activation of apoptosis mechanism in cancer cells. Western blotting analysis also showed the activation of initiator caspase-8 and caspase-9, and executor caspase-3 in GBM 8401 cells. Pretreatment of a caspase-8 inhibitor (Z-IETD-FMK), caspase-9 inhibitor (Z-LEHD-FMK) and caspase-3 inhibitor (Z-DEVE-FMK) significantly increased the viable of GBM 8401 cells implied that the participations of caspases. Western blots indicated that the protein levels of Caspase-3, -8, -9, Bax, Bid, AIF, Endo G, and PARP that were increased and Bcl-2, Bcl-xLthat were decreased may have led to apoptosis. Danthron induced apoptosis through the intrinsic caspase-dependent and -independent pathways in GBM 8401 cells. This research has demonstrated that Danthron induces cytotoxicity and apoptosis in GBM 8401 cells through mitochondria-related, caspase-dependent and -independent pathways. Danthron may be further evaluated as a chemotherapeutic agent for human brain cancer