지모 추출물이 난소암 세포 성장 저해 및 종양 성장 억제에 미치는 영향

Abstract

Objectives: Anemarrhena asphodeloides Bunge is a dried medicinal herb with anti-inflammatory, anti-oxidant and anti-cancer effects on colon cancer, breast cancer, and pancreatic cancer. However, since research on Anemarrhena asphodeloides Bunge’s ovarian cancer is lacking, the anti-cancer effect of Anemarrhena extract on ovarian cancer cells was evaluated through in vitro and in vivo experiments. Methods: Anemarrhena asphodeloides extracts used were water extract (AAW) and ethanol extract (AAE). Normal Human Dermal Fibroblast (NHDF) cells, a normal cell line, and SKOV3 and A2780 cells, epithelial ovarian cancer cells, were treated with various concentrations (12.5, 25, 50, and 100 μg/ml) of AAW and AAE. The toxicity to normal cells and the viability of ovarian cancer cells were analyzed using the WST-8 assay. To confirm the effect of AAE on the growth inhibition and apoptosis of ovarian cancer cells, anchorage-independent cell growth experiment (soft agar assay), cell cycle analysis, apoptosis analysis, and Western blot analysis were performed. In vivo experiments were performed using a xenograft model to evaluate the effect of AAE on the inhibition of tumor growth of ovarian cancer. Results: Our experimental results showed that AAW and AAE did not exhibit toxicity in NHDF cells, and significantly reduced cell viability in SKOV3 and A2780 cells in a concentration- and time-dependent manner. Morphological changes in ovarian cancer cells after AAW and AAE treatment were observed. As the treatment concentration and time increased, cell size decreased and cells shrank or changed in shape. Anchorage-independent cell growth experiments in SKOV3 and A2780 cells showed that the number and growth of colonies were significantly reduced by AAE treatment. To evaluate the effect of AAE on the cell cycle, cell cycle analysis was performed using flow cytometry, and the results showed that AAE induced cell cycle arrest in the G2/M phase. In addition, Annexin V/PI staining was performed to evaluate whether apoptosis was induced, and AAE promoted apoptosis in SKOV3 and A2780 cells in a concentration-dependent manner. After tumors were formed by subcutaneous injection of human ovarian cancer cells KOV3 into immunodeficient BALB/c nude mice, AAE was administered intraperitoneally for 6 weeks, and it showed a significant effect of inhibiting tumor growth. Conclusions: These in vitro and in vivo experimental results suggest that Anemarrhena asphodeloides extracts has anticancer effects that inhibit the growth and proliferation of ovarian cancer cells, induce apoptosis, and suppress tumor growth.