摘要
目的研究构树绿原酸样化合物诱导胃癌SGC-7901细胞凋亡的作用及机制。方法采用MTT法检测构树绿原酸样化合物对SGC-7901细胞增殖的影响,DAPI染色法观察细胞形态,Annexin V/PI双染色法结合流式细胞术检测细胞凋亡,PI染色结合流式细胞术检测细胞周期,DCHF-DA探针荧光显微镜观察细胞内活性氧(ROS)变化,JC-1染色荧光显微镜观察细胞线粒体膜电位变化,Westernblotting检测细胞p53、Bcl-2、Bax、CytochromeC、p-p38、p-JNK、JNK、细胞外调节蛋白激酶(ERK)、p-ERK蛋白表达量。结果构树绿原酸样化合物能显著抑制SGC-7901细胞增殖,且呈时间和剂量依赖性;给药组细胞内出现染色质浓缩和凋亡小体,细胞周期被阻滞在G2/M期,且线粒体膜电位显著下降(P<0.05、0.01),ROS水平显著升高(P<0.05、0.01)。构树绿原酸样化合物能显著上调SGC-7901细胞p53、Bax、Cytochrome C和p-p38蛋白表达水平(P<0.05、0.01、0.001),显著下调p-ERK和Bcl-2表达水平(P<0.01、0.001)。结论构树绿原酸样化合物可能通过p38-MAPK和ERK-MAPK信号通路介导线粒体氧化应激途径诱导胃癌细胞SGC-7901凋亡。
Objective To explore the mechanism of gastric carcinoma cell SGC-7901apoptosis induced by.chlorogenic acid-like compounds extracted from Broussonetia papyrifera (CALCBP)bark.Methods The SGC-7901cells were used to evaluate the- anti-tumour activity of the extract in vivo,and the proliferation of cells was examined by MTT assay.The cell morphological changes of cells were observed by DAPI staining;The cell apoptosis and the cell cycle were detected respectively by flow cytometry after PI and Annexin V/PI staining;The intracellular ROS were determined under the fluorescence microscope using DCHF-DA probe,the changes of mitochondrial membrane potential were observed by JC-1staining.The protein expression of p53,Bcl-2,Bax,and Cytochrome C,,p-p38,p-JNK,JNK,p-ERK,ERK were analyzed by Western blotting.Results The proliferation of SGC-7901cells was inhibited significantly by CALCBP in dose-dependent and time-dependent manner,the condensed chromosome and apoptotic body can be observed in the treated cells and the cell cycle was arrested in G2/M phase,the mitochondrial membrane potential was significantly decreased,whereas the cellular ROS levels of the treated cells were significantly increased.Moreover,the protein expression of p53,Bax,Cytochrome C,and p-p38were significantly up-regulated and p-ERK and Bcl-2expression were significantly down:regulated.Conclusion The apoptosis of gastric cancer cell SGC-7901induced by CALCBP was probably related to oxidative stress of the cell mitochondrial via p38-MAPK and ERK-MAPK signal pathways.
作者
朱泽欣
赵云涛
陈绍红
刘艳芬
刘铀
ZHU Ze-xin;ZHAO Yun-tao;CHEN Shao-hong;LIU Yan-fen;LIU You(College of Agricultural,Guangdong Ocean University,Zhanjiang 524088,China;College of Food Science and Technology,Guangdong Ocean University,Zhanjiang 524088,China)
出处
《中草药》
CAS
CSCD
北大核心
2018年第22期5345-5351,共7页
Chinese Traditional and Herbal Drugs
基金
广东省农业科技计划项目"构树综合加工利用技术研究"(0809035)
关键词
构树
绿原酸样化合物
细胞凋亡
氧化应激
丝裂原活化蛋白激酶信号通路
Broussonetiapapyrifera(Linn.)L'Her.ex Vent.
chlorogenic acid-like compounds
cell apoptosis
oxidative stress
MAPK signal pathway
