이종이식 누드마우스 종양 모델을 이용한 전호 수용성 추출물의 항암효과 분석

Abstract

Objective : The anti-cancer effect of Anthriscus sylvestris Hoffmann aqueous layer (ASAL) investigated by subcutaneous injection of KB human oral cancer cell and A549 human lung cancer cell of xenograft nude mouse models.

Materials and Methods : Cell viability and cytotoxicity effects were evaluated by MTT assay after ASAL treatment for 24 h. For western blot analysis, primary antibodies such as phospho-p53 (Ser 15), phospho-ERK 1/2, EGFR, phospho-EGFR (Tyr 992), phospho-EGFR (Tyr 1045), phospho-EGFR (Tyr 1068), Bcl-2, cleaved caspase-3, cleaved caspase-7, cleaved caspase-8, cleaved caspase-9 and poly ADP ribose polymerase (PARP) were used in vitro and in vivo. KB oral cancer cells and A549 lung cancer cells cultured and subcutaneous injected them to nude mouse. ASAL was administrated to xenograft nude mouse tumor model orally every day. It has been raised and treated up to 17 days and 30 days. Their body weight, tumor weight and tumor volumes were measured every other day a week. Hematology and serum biochemistry were performed for the safety assessment of ASAL in vivo.

Results : ASAL significantly inhibited cell growth in KB, FaDu oral cancer cell lines and A549 lung cancer cells in dose-dependent manners without toxicity in NIH/3T3 fibroblast and primary rat chondrocyte cells. ASAL-treatment cells showed inhibition of ERK 1/2 phosphorylation in the KB and FaDu oral cancer cell lines. It also significantly inhibited EGFR phosporylation (Tyr 1068) in the A549 lung cancer cells. Moreover, ASAL inhibited protein expression level of anti-apoptotic factor Bcl-2. In contrast, activated cleaved caspase-3, cleaved caspase-9 and poly ADP ribose polymerase (PARP) in the KB and A549 cells. KB subcutaneous xenograft nude mouse tumor models (ASAL ; 400 ㎎/kg) and A549 subcutaneous xenograft nude mouse tumor models (ASAL ; 80 ㎎/kg, ASAL ; 200 ㎎/kg) ASAL treatment groups of were suppressed 40%, 45.6% and 59.4% tumor inhibition rate compared to non-treatment group, respectively. In addition, tumor tissues of ASAL treatment groups, poly ADP ribose polymerase (PARP) cleavage was significantly increased compared to non-treatment groups. No significant differences in body weight, hematology and serum biochemistry between control and ASAL (500 ㎎/kg) group were found.

Conclusion : These results indicated that ASAL may be suppress of cell proliferation and subcutaneous xenograft nude mouse tumor growth by inducing the intrinsic and extrinsic apoptotic pathway. Furthermore, it can be a basis development of water soluble anti-cancer drug.