Studies on the Mechanisms of Apoptosis induced by the Methanol Extract of A.camphorate in A549 cells.

• Main Authors: Chiuan-Shiang Shu, 許筌翔
• Other Authors: Tai-An Chiang Ph.D.
• Format: Others
• Language: zh-TW
• Published: 2014
• Online Access: http://ndltd.ncl.edu.tw/handle/20001627484830002276

碩士 === 中華醫事科技大學 === 醫學檢驗生物技術系碩士班 === 102 === Backgrounds: Lung cancer is the first cause of death in men and women all over the world. It is also the first cause of death induced by cancer in Taiwan. There are about 22 people died for lung cancer per day, where 19 of 22 is the result of non-small cell lung cancer. Non-small cell lung cancer includes Adencarcinoma,quamous cell, and large cell carcinoma, and it accounts for about 85 % of lung cancer cases. It had been known for many years that selected mushrooms are effective in fighting against cancer in stomach, esophagus, lungs and others, whereas the mechanisms of anticancer therapeutic effects of mushrooms are poorly understood. Antrodia camphorata is a cultured volvatus mushroom in Taiwan. Bioactive effects in A.camphorate have been the focus of recent research on antitumor, and anticancer activity, and also that the treatment of various diseases, such as liver disease, cancer,poisoning, abdominal pain, and apply on clinical medicine. Our previous study demonstrated that the methanol extract from A. camphorate has induction activity of apoptosis in MCF-7 breast cells. It was suggested that MEAC may be a potential chemo preventive agent on apoptosis. The aim of the present study was to investigate the mechanisms of apoptosis induced by MEAC in A549 cells. Materials and Methods: Cytotoxicity of different concentration (0 ~ 40 μg / ml) of MEAC has been measured by MTT cytotoxicity analysis. We further verify the mechanism of MEAC on cytotoxicity in human lung adenocarcinoma A549 cells with flow cytometry analysis and Western blotting analysis. Results: Our data showed that cytotoxic activity induced by MEAC in a dose-response manner. Therefore, we chose 15, 20, 25, 30μg/ml of MEAC to do the experiments without affecting the results. The result of flow cytometry analysis has been demonstrated that cell apoptosis was also significantly induced by MEAC in a dose-response pattern. Moreover, western blotting assay showed that cytochrome C, Caspase-9, Caspase-3, Caspase-7, 89 kDa PARP (poly ADP-ribose polymerase) protein expression increased with the increasing concentration of MEAC. Furthermore, BCL-XL protein expression performance will depend on the amount of increase in the reducing concentration of MEAC. Collectively, the mechanism of apoptosis induced by MEAC in A549 cells may be partially mediated through induction of Caspase protein expression and inhibition of BCL-XL protein expression. Conclusion: The study had shown that MEAC through induction of apoptosis pathway, then inhibit the growth of A549 cancer cells. So, MEAC may be a highly value product with anti-cancer drugs.