丹皮酚诱导人食管癌Eca-109裸鼠移植瘤凋亡的机制探讨

目的通过观察用药后COX-2、Bcl-2和Survivin的变化,探讨丹皮酚(paeonol,Pae)诱导Eca-109食管癌裸鼠移植瘤凋亡的机制。方法体外培养食管癌Eca-109细胞,裸鼠皮下接种Eca-109细胞建立裸鼠移植瘤动物模型,36只荷瘤裸鼠随机分为6组,分别为模型对照组、Pae不同剂量组(25、50、100、200mg·kg-1)和阳性药对照组(cisplatin,CD-DP,5mg·kg-1)。治疗2wk后处死裸鼠,剥取瘤体称瘤重并计算抑瘤率。用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(TUNEL)法检测肿瘤细胞凋亡。免疫组化S-P法检测移植瘤组织COX-2、Bcl-2和Survivin的表达。结果Pae50、100和200mg·kg-1组和CDDP5mg·kg-1组均能明显抑制裸鼠皮下肿瘤的生长,抑瘤率分别为23·54%、27·91%、34·46%和58·71%,与模型组比较差异均有显著性(P<0·05orP<0·01)。TUNEL染色可发现棕褐色的凋亡细胞呈散在或片状分布,Pae各剂量组的凋亡指数(apoptosis index,AI)分别为(11·02±2·58)%、(19·80±2·77)%、(24·48±4·35)%和(27·13±4·39)%,与模型组(4·81±0·83)%比较,差异均有显著性(P<0·05orP<0·01)。免疫组化结果显示,Pae能明显抑制移植瘤组织COX-2、Bcl-2和Survivin的表达(P<0·05 or P<0·01)。结论Pae能抑制Eca-109食管癌裸鼠移植瘤生长、诱导凋亡而发挥抗肿瘤作用,其机制可能与下调COX-2的表达并抑制Bcl-2和Sur-vivin的表达有关。

Investigation of apoptosis mechanism of Pae on human esophageal cancer Eca-109 cell carcinoma xenograft in nude mice

Aim To investigate the mechanism of apoptosis on human esophageal cancer Eca-109 cell carcinoma xenograft in nude mice induced by paeonol(Pae)by detecting changes of expressions of COX-2,Bcl-2 and Survivin.Methods Human esophageal carcinoma Eca-109 cells were cultured in vitro. After the nude mice model of the subcutaneous transplanting tumor was established by human esophageal cancer Eca-109,the nude mice were randomly divided into six groups:various dosages of Pae treated groups(25,50,100,200 mg·kg-1),cisplatin(CDDP)positive control group(5 mg·kg-1)and model group.After two weeks of treatment,mice were killed and the tumors were taken out to weigh.The tumor inhibitory rate was calculated. Cell apoptosis in situ was examined by a TUNEL assay in the samples of carcinoma. Immunohistochemistry(S-P)was used to examine the expressions of COX-2,Bcl-2 and Survivin.Results The growth of implanted tumor was markedly inhibited in Pae groups(50,100,200 mg·kg-1)and CDDP group(5 mg·kg-1),the inhibitory rate being 23.54%,27.91%,34.46%,58.71% respectively(P<0.05 or P<0.01).It was found that the apoptotic cells which stained yellow distributed scatterly or diffusely by TUNEL techniques.The apoptosis indexes of Pae groups were (11.02±2.58)%,(19.80±2.77)%,(24.48±4.35)% and(27.13±4.39)% respectively,which were significantly higher than (4.81±0.83)% in control group(P<0.05 or P<0.01).The expressions of COX-2,Bcl-2 and Survivin in Pae groups were significantly lower than those of the control group detected by immunohistochemistry (P<0.05 or P<0.01).Conclusion Pae could inhibit the tumor growth and induce apoptosis of human esophageal carcinoma Eca-109 in vivo,the mechanism of which might be related with down-regulation of COX-2,Bcl-2 and Suvivin.