Cinnamomum cassia Presl crude extract and its major component α-phellandrene induce apoptosis in leukemia cell lines in vitro and the promote immune response in leukemia mice in vivo

• Main Authors: Jen-Jyh Lin, 林楨智
• Other Authors: Jaung-Geng Lin
• Format: Others
• Language: zh-TW
• Published: 2014
• Online Access: http://ndltd.ncl.edu.tw/handle/bzg67h

博士 === 中國醫藥大學 === 中醫學系博士班 === 102 === Although reports have shown that α-phellandrene (α-PA) is one of the monoterpenes and is often used in the food and perfume industry. However, there is no available information to show that α-PA induced cell apoptosis in cancer cells, thus, we investigated the effects of α-PA on the cell morphology, viability, cell cycle distribution and apoptosis in mice leukemia WEHI-3 cells in vitro. Results indicated that α-PA induced cell morphological changes and decreased viability, induced G0/G1 arrest and sub-G1 phase (apoptosis) in WEHI-3 cells. α-PA increased the productions of reactive oxygen species (ROS) and Ca2+ and decreased the levels of mitochondrial membrane potential (ΔΨm) in dose- and time- dependent manners in WEHI-3 cells that were analyzed by flow cytometer. Results from confocal laser microscopic systems examinations show that α-PA promoted the releases of cytochrome c, AIF and Endo G from mitochondria in WEHI-3 cells. These results are the first findings to provide new information for understanding the mechanisms by which α-PA induces cell cycle arrest and apoptosis in WEHI-3 cells in vitro. On the other hand, we investigated the effect of α-PA in the DNA damage and repair of murine leukemia cell-associated protein expression. We have used comet assay, DAPI staining and DNA gel electrophoresis to examine the DNA damage and condensation. The result showed that α-PA-induced DNA damage and condensation in a concentration-dependent manner. In addition, Western blot analysis to measure DNA damage and repair-related changes in protein expression, the results indicated that increased 14-3-3 ?? , P-P53, P-H2A.X and MDC1 protein expression, but inhibition of p53 MGMT, DNA-PK and BRCA - 1 protein. α-PA-induced DNA damage and condensation is a concentration – dependent manner. Effects of α-PA promote immune responses in normal BALB/c mice and immune responses on a mouse model of leukemia were examined. α-PA increased the percentage of CD3 (T-cell maker), CD11b (monocytes) and the Mac-3 (macrophages) but decreased the percentage of CD19 (B-cell maker) cell surface markers in α-PA-treated groups as compared with the untreated included oliver oil treated groups. α-PA increased B- and T- cell proliferation at 25 mg/kg treatment. α-PA 25 mg/kg treatment might exhibit promoted immune responses in a murine in vivo.