摘要
目的:研究西黄丸对人结肠癌细胞SW480迁移及侵袭的影响,并初步探讨其作用机制。方法:以人结肠癌细胞SW480为研究对象,细胞划痕试验检测不同剂量西黄丸含药血清对SW480细胞迁移的影响,并Transwell侵袭实验检测西黄丸含药血清对细胞侵袭能力的改变;用Western blot的方法检测细胞外调节蛋白激酶(ERK)和p-ERK蛋白表达水平,RTq PCR法检测E-钙黏蛋白(E-cadherin),多聚腺苷二磷酸核糖聚合酶1(PARP-1)、波形蛋白(Vimentin)和转录因子Snail的mRNA水平。结果:与空白组比较,西黄丸含药血清可降低SW480细胞迁移及侵袭的能力,增加E-cadhenrin mRNA表达水平,并抑制PARP-1,Vimentin和Snail mRNA表达水平,明显降低ERK蛋白的磷酸化水平,均具有统计学意义(P<0.05)。结论:西黄丸含药血清可抑制SW480细胞的体外侵袭及迁移,并影响上皮-间皮转化(EMT)相关基因E-cadherin,PARP-1,Vimentin和Snail的mRNA表达,其机制可能与ERK/MAPK信号通路有关,本研究为西黄丸抑制结肠癌转移提供了科学证据。
Objective：To investigate the effects and potential mechanisms of Xihuang Wan on cell migration and invasion in SW480 human colorectal cancer cells.Method：SW480 human colorectal cancer cells were used as the cell model.Wound healing and transwell assay were used to determine the inhibitory effects of serum of rats containing Xihuang Wan on cell migration and invasion in SW480 cells,respectively.Western blot was used to measure the protein expression levels of ERK and phospho-ERK,and the qPCR assay was used to evaluate the mRNA expression levels of E-cadherin,PARP-1,Vimentin and Snail in SW480 cells after serum containing Xihuang Wan treatment.Result：The abilities of cells migration and invasion showed dose-dependent reducing in SW480 cells after treatments with the serums containing Xihuang Wan.The qPCR results showed that the serum containing Xihuang Wan enhanced E-cadherin,while reduced PARP-1,Vimentin and Snail mRNA expression levels.In addition,Western blot results showed the decreasing phosphorylation of ERK after treatments with serum containing Xihuang pill.Conclusion：The serums containing Xihuang Wan inhibited cells migration and invasion,affected the mRNA expression levels of epithelial-mesenchymal transiton （EMT） related markers including E-cadherin,PARP-1,Vimentin and Snail in SW480 cells.The mechanism may be related to the ERK/ MAPK pathway.Our studies provide further pharmacological basis for developing Xihuang Wan as a preventive/ chemotherapeutic agent of metastasis in colorectal cancer.
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2015年第2期136-141,共6页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81302892)
广东省自然科学基金项目(S2013040016226)
中国博士后基金项目(2013M530363)
关键词
西黄丸
含药血清
结肠癌
肿瘤转移
上皮-间皮转化
Xihuang Wan
drug serum
colorectal cancer
metastasis
epithelial-mesenchymal transiton
