Evaluation of the Inhibitory Activity of Spiranthes sinensis (Per.) Ames Crude Extracts on Breast Cancer Cells

• Main Authors: Liao, Po-Chieh, 廖伯杰
• Other Authors: Kao, Chien-Yuan
• Format: Others
• Language: zh-TW
• Published: 2019
• Online Access: http://ndltd.ncl.edu.tw/handle/szhev3

碩士 === 國立宜蘭大學 === 生物資源學院碩士在職專班 === 107 === Spiranthes sinensis, local name ‘Shou Cao”, is a traditional Chinese herbal medicine. The whole plant can be used as a medicine. The wild population of this plant is not easy to find. The Spiranthes sinensis (Spiranthes sinensis (Per) Ames; Ss) mature plant used in this study are derived from in vitro propagation. As a result, the source of the plant material is stable and the traits are consistent. There are few studies on the inhibitory activity of breast cancer cells by Spiranthes sinensis extract. This study is the first attempt to use 4T1 breast cancer cells and anti-oxidation experiments to evaluate the ability of Ssextract to inhibit cancer cells. Also the North American Shou Cao (Spiranthes cernua; Sc) is used in the same experiments to compare with Ss in term of their inhibitory effects. Water and 95% alcohol extraction methods were applied to the whole plant and root of the Ss and scseparately. After applying the extracts at concentrations of 50 μg/ml, 100 μg/ml, 200 μg/ml, and 400 μg/ml to 4T1 breast cancer cells for 24 hours, the results showed that the alcohol extract of Ssinhibited the activity of breast cancer cells significantly. At a concentration of 100 μg/ml, the survival rate of breast cancer cells is reduced to 26.3%±0.8, which is significantly better than that ofSc 91.6%±2.2. In contrast, the water extracts of these Ssand Sc, even at the highest concentration of 1 mg/ml, have poor inhibitory ability against cancer cells, which in dicates that the alcohol extract of Sscancer inhibition capacity is significantly higher than water extracts. In terms of plant portion used for extraction, the whole plant is superior to the root. The ability of Sc to inhibit cancer cells in both alcohol or water extracts is not as good as Ss; Further experiments on DPPH and ABTS+ showed that the ability of Ssextract to scavenge free radicals and inhibit breast cancer ROS was superior to that of Sc. Furthermore after 16 hrs treatment at concentrations of 50 μg/ml and 75 μg/ml alcohol extract, the breast cell viability of the Ss treatment was 76%±2.2, 67%±1.7 respectively, in contrast to Sc treatment group, 97% ± 2.6, 94% ± 1.98 of breast cancer cells survived. Gelatin zymography analysis showed that the MMP-9 zymogen activity of breast cancer cells decreased with the increase of Ss extract concentration added.AlsoWestern blot clearly showed that the active MMP-9 protein is significantly inhibited by 75 μg/ml of the Ss alcohol extract. The inhibitoryeffect towards breast cancer cell by Ss alcohol extract were also evident at a concentration of 75 μg/ml through cell wound healing test. Summarizing the results of this study, the alcohol extract ofSshas the potential to become a drug of anti-cancer or adjuvant treatment of cancer.