Investigation of the mechanisms of photodynamic therapy with Lonicera japonica-induced apoptosis in human lung squamous carcinoma CH27 cells

• Main Authors: Meng-Yi Wang, 王孟宜
• Other Authors: Hong-Zin Lee
• Format: Others
• Language: zh-TW
• Published: 2007
• Online Access: http://ndltd.ncl.edu.tw/handle/42602912413878927169

碩士 === 中國醫藥大學 === 藥學系碩士班 === 95 === Lonicera japonica T. (Caprifoliaceae) has been used in Chinese medicine for a long time and used as a healthy food in recent years. The active components from the flowers and leaves of Lonicera japonica have been reported to exhibit antiproliferative effects and induce apoptosis in some tumor cells. In the present study, Lonicera japonica-mediated photodynamic therapy (PDT) has been shown to induce apoptosis in CH27 lung carcinoma cells. During Lonicera japonica-mediated PDT-induced a typical apoptosis, the exposure of the membrane phosphatidylserine to the external cellular environment, apoptotic bodies and DNA condensation were observed. Photodynamic therapy is becoming widely accepted as a potential treatment for many forms of cancer. It has been demonstrated that PDT is a curative treatment for the patients of non-small cell lung carcinoma who are not able to undergo surgery or radiotherapy. Therefore, the major purpose of this study was to characterize the effect and mechanisms of PDT with Lonicera japonica-induced CH27 lung cancer cell apoptosis. This study has demonstrated that Lonicera japonica is a photosensitizing agent and induces CH27 cell death after 4 h treatment. The changes in protein expression of Bcl-2, Bax, AIF and caspase-3 are involved in PDT with Lonicera japonica-induced CH27 cell apoptosis. It is noteworthy that PDT with Lonicera japonica induced a marked decrease in the mRNA level and protein activity of caspase-3. The increase in the activity of matrix metalloproteinase-2 and -9 was also observed in this study. Since PDT with Lonicera japonica induced the changes in morphology and F-actin localization in CH27 cells, we used the immunostaining to demonstrate HSP27 and p38 protein involvement in PDT with Lonicera japonica-induced CH27 cell apoptosis. Luteolin was used as a quality control of alcohol extract of Lonicera japonica in HPLC analysis. The HPLC data showed that luteolin is the component in the Lonicera japonica, consisting of approximately 0.13% of Lonicera japonica. We have also demonstrated that luteolin and chlorogenic acid are not the photosensitizer in Lonicera japonica. Ethyl acetate extracts of Lonicera japonica may be the bioactive layer of Lonicera japonica in PDT with Lonicera japonica-induced CH27 cell apoptosis.