The crude extracts and active ingredients from Tetrastigma hemsleyanum on anti-cancer effects

• Main Authors: De-Min, Wu, 吳德珉
• Other Authors: Hui-Fen, Liao
• Format: Others
• Language: zh-TW
• Published: 2018
• Online Access: http://ndltd.ncl.edu.tw/handle/z48a8c

碩士 === 國立嘉義大學 === 生化科技學系研究所 === 106 === Tetrastigma hemsleyanum is a genus of plants in the grape family, Vitaceae. The roots of T. hemsleyanum are used as a folk medicine in Asia for the anti-inflammatory, analgesic, and antipyretic properties. The major active ingredients, rutin and kaempferol, were reported with the activities of anti-oxidation, anti-inflammatory, and enhancing the inhibition of cancer cells. However, the exact anti-cancer activities of the crude extracts and active compounds from T. hemsleyanum are still unclear. Lung cancer, also known as lung carcinoma, is a malignant lung tumor characterized by uncontrolled cell growth in tissues of the lung. The growth can spread beyond the lung by the process of metastasis into nearby tissue or other parts of human body. Lung cancer is the top ten cancer rankings in Taiwan and the leading cause of death worldwide. Of these deaths, nearly 60% of patients progress into advanced stages with metastasis. Therefore, new strategies for enhancing the treatment efficiency of lung cancer are necessary. The aim of this study explored the extraction by H2O and EtOH from T. hemsleyanum, and assayed the antioxidant activity, total phenolic and flavonoid content from the extracts, as well as investigation of the active integrates on anti-cancer effects. The results demonstrated that EtOH extracts from T. hemsleyanum have better antioxidant activities by 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay, while H2O extracts have higher phenolic and flavonoid contents, indicating that there are many components contained in the T. hemsleyanum with various activities. The known active compounds rutin and kaempferol were further analyzed. Cell viability by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed that kaempferol was effective in inhibition of both human A549 and murine LLC1 lung cancer cell lines. However, rutin did not significantly inhibit the growth of A549 and LLC1 cells at the concentrations ranging from 50 to 400 μM. In addition, the combination of kaempferol and paclitaxel enhanced the inhibition of both cell lines. At the concentration of 50 μM, kaempferol combined with paclitaxel (50 μM) increased the inhibition of cell growth by 11% and 30% in A549 and LLC1 cells, respectively, as compared to paclitaxel (50 μM) alone. In cell cycle analysis, kaempferol induced the cell cycle arrest at the S phase in A549 cells, paclitaxel increased the sub-G1 and S phases, and the combined treatment significantly upregulated the S and G2/M arrest. After treatment with paclitaxel, kaempferol alone, or combinational treatment in LLC1 cells, all of them showed no significant change of cell cycle. Kaempferol alone and combinational treatment promoted the G2/M phase in LLC1 cells, indicating that kaempferol arrested the cell cycle at G2/M phase. The reason that paclitaxel effect on LLC1 cells is not sensitive still needed to be further clarification. In summary, the present study demonstrated that the H2O and EtOH extracts from T. hemsleyanum have different antioxidant activities, total phenolic, and flavonoid content. The active ingredient kaempferol has a higher growth inhibitory effect on lung cancer cells than rutin. The combination of kaempferol with paclitaxel has an enhancing effect on anti-lung cancer activity