The aim of this study was to examine the effects of aqueous crude extract of Acanthus ebracteatus Vahl. (AE) on tumor growth and tumor angiogenesis in human cervical cancer cells contained human papillomavirus (HPV)-16 DNA-implanted nude mice model. In vitro the growth inhibitory effect of AE was studied by using cell types including CaSki and Human dermal fibroblast cells, HDFs. The cell growth-inhibitory activity and IC₅₀ of AE were determined at different incubation time 24-, 48-, and 72-hours by using Trypan blue exclusion method and MTT assay. Under in vitro condition, AE exhibited weak anti-proliferative effect on CaSki, cell growth in a dose-dependent manner with exposure time at 48-hr. AE could inhibit the growth of CaSki cells, the only one cancer cell, while less inhibitory effect to normal cells, HDFs (p<0.05). In vivo study Female BALB/c nude mice (weighing 20-25 g, age 5-6 weeks) were used for establishment of HPV 16-positive cervical cancer mice model. CaSki cells, 1x10⁷ cells/MEM 200{u1D707}l were injected subcutaneously in the middle dorsum of each animal (HPV group). One week after, mice were fed orally with AE 300 or 3,000 mg/kg BW/day for 14 and 28 days (HPV-AE groups), distilled water were used as vehicle control. After 14- and 28- day of treatments, the tumor microvasculature and capillary vascularity (CV) were determined by using laser scanning confocal microscopic system. Tumor tissue sample of each mouse was collected for immunohistochemical examination of biomarkers, vascular endothelial growth factor (VEGF) and p53 proteins. In vivo study, the increasing in tumor volume were observed during day 14th – 28th, and high dose treatment of AE (3,000 mg/kg BW) could inhibit the increase of tumor volume (P<0.001). A large number of microvascular network around tumor area was observed in HPV-Veh groups on day 21st and 35th after cancer-cell inoculation. Tumor capillary vascularity (CV) in HPV-Veh groups was significantly increased when compared to Con-Veh group (P<0.001). High dose treatment of AE significantly attenuated the increase of tumor angiogenesis on both 14- and 28-days of treatment (P<0.001). Tumor VEGF expression with strong intensity was detected in HPV-Veh group, and %area of VEGF expression in HPV-3,000 AE both 14-and 28-days were significantly less than HPV-Veh and HPV-300 AE group (P<0.001). Tumor p53 expression in HPV-Veh group was detected with a very small number or almost none. The elevation of p53 expression in HPV-AE groups was observed in dose dependent manner (P<0.001). Our novel findings demonstrated that aqueous crude extract of Acanthus ebracteatus Vahl. Could inhibit tumor growth and tumor angiogenesis in cervical cancer mice model. Therefore, it provides the research evidence that Acanthus ebracteatus Vahl. Might be contribute to the development of new therapeutic management against cervical cancer in the near future.
