摘要
目的观察吉祥草中甾体皂苷RCE-4对宫颈癌裸鼠移植瘤的抑制作用。方法采用传统方法分离鉴定RCE-4。将0.3 m L Caski(4×107/m L)细胞悬液接种于裸鼠右侧后腿背部皮下,建立宫颈癌移植瘤模型,然后按瘤体积大小随机分为模型组、紫杉醇(10 mg/kg)和RCE-4(25、50、100 mg/kg)组,1次/d给药,连续4周,末次给药的次日处死裸鼠,称量裸鼠体质量、瘤质量,测量肿瘤体积,计算抑瘤率。光镜下观察移植瘤组织形态学变化,TUNEL染色测定细胞凋亡率,免疫组化检测肿瘤组织中COX-2表达,荧光定量PCR测定移植瘤组织中Bcl-2、Bax、Caspase-3和Caspase-9 mRNA表达,计算Bcl-2/Bax值,Western blot检测移植瘤组织中Survivin蛋白表达。结果 RCE-4(25、50、100 mg/kg)可明显降低宫颈癌裸鼠移植瘤的体积和瘤质量(P<0.05,P<0.01),上调Bax、Caspase-3和Caspase-9 mRNA表达,下调Bcl-2 mRNA表达和COX-2、Survivin蛋白表达及Bcl-2/Bax值(P<0.05,P<0.01)。结论RCE-4对宫颈癌裸鼠移植瘤的生长具有显著抑制作用,其机制可能与其上调Bax、Caspase-9、Caspase-3的表达和下调Survivin、Bcl-2、COX-2的表达有关。
Objective To determine the inhibitory effect of steroid sapogenin RCE-4 derived from Reineckia carnea （Andr.） Kunth on the growth of human cervical cancer xenograft in nude mice. Methods Steroidal saponin RCE-4I was extracted and identified by conventional methods. Nude mice model of transplanted tumor of human cervical cancer Caski cells were established by subcutaneously injection of 0.3 mL cells （4×107） under the right hind leg. Caski cervical cancer model mice were randomly divided into model group, paclitaxel （10 mg/kg） and RCE-4 （25, 50 and 100 mg/kg） groups, once a day for 4 weeks. Then all mice were sacrificed in 24 h after the final medication, and the volume and weight of transplanted tumor masses were measured to calculate the tumor inhibitory rate. Morphological changes of transplanted tumor tissue were observed by light microscopy, apoptotic rate was determined by TUNEL staining, COX-2 expression in the tumor tissues was detected by immunohistochemical assay, the mRNA levels of Bcl-2, Bax, Caspase-3 and Caspase-9 were detected by real-time PCR, and the Bcl-2/Bax ratio was analyzed, and the expression level of survivin protein in the tumor tissue was detected by Western blot analysis. Results RCE-4 （25, 50 and 100 mg/kg） significantly inhibited the volume and weight of the transplanted tumor in the nude mice （P〈0.05, P〈0.01）, up-regulated the mRNA expression levels of Bax, Caspase-3 and Caspase-9, down-regulated the expression of Bcl-2 at mRNA level and COX-2 and survivin at protein level, and reduced the Bcl-2/Bax ratio （P〈0.05, P〈0.01）. Conclusion RCE-4 significantly inhibites the growth of human cervical cancer in nude mice, which may be associated with up-regulating the expression of Bax, Caspase-3 and Caspase-9, and down-regulating the expression of survivin, Bcl-2 and COX-2.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2016年第5期476-482,共7页
Journal of Third Military Medical University
基金
国家自然科学基金面上项目(21272136)
湖北省自然科学基金(2013CFB224)
宜昌市科学技术局项目(A13-302a-14)
三峡大学培优基金(2014PY071)~~
关键词
甾体皂苷RCE-4
吉祥草
宫颈癌
CASKI细胞
裸鼠
细胞凋亡
steroidal saponin RCE-4
Reineckia carnea （Andr.） Kunth
cervical cancer
Caski cells
nude mice
apoptosis
