摘要
目的:探讨白扁豆多糖对人胃癌细胞凋亡的作用及其相关机制。方法:胃癌细胞HGC-27和SGC-7901经终浓度为16、8、4、2、1和0μg/ml的白扁豆多糖作用24、48和72h,各设3个复孔。MTS法检测其增殖活性;分别取经4、0μg/ml白扁豆多糖作用24h的HGC-27和SGC-7901细胞(各3个复孔),JC-1染色观察线粒体膜电位,流式细胞仪分析细胞周期和凋亡率,QPCR法探讨Bcl-2、caspase-3和Bax基因的mRNA转录水平。结果:白扁豆多糖作用后,HGC-27和SGC-7901细胞线粒体膜电位降低;细胞增殖显著受抑制(P<0.01),且效果与药物作用浓度和时间有关;细胞凋亡率分别为53.15%和38.77%,均较PBS处理组(8.07%和6.03%)明显增加(P<0.01),而细胞周期无显著变化;同时,细胞内Bcl-2基因的转录水平明显受抑制,Bax和caspase-3基因的转录明显上调(P<0.01)。结论:白扁豆多糖可通过调节Bax-Bcl-2-caspase3通路,诱导胃癌细胞HGC-27和SGC-7901凋亡。
Objective: To investigate the effect of white lentil polysaccharides on the apoptosis of human gastric cancer cells and its related mechanism. Methods: Gastric cancer cell lines HGC-27 and SGC-7901 were treated with 16,8,4,2,1 and 0 μg/ml white lentil polysaccharides for 24,48 and 72 hours respectively,each experiment 3 wells. The proliferation activities of gastric cancer cells were detected with Cell Titer 96-?AQueous Non-Radioactive Cell Proliferation Assay( MTS). HGC-27 and SGC-7901 cells were treated with 0 or 4 μg/ml white lentil polysaccharides for 24 hours respectively,each experiment 3 wells,then the changes of cell mitochondrial membrane potential were observed by JC-1 staining,cell cycle and cell apoptosis rates were evaluated with flow cytometry,the levels of mRNA expression of Bcl-2,caspase-3 and Bax genes were investigated with real-time quantitative PCR detecting system( QPCR). Results: After the gastric cancer cell lines HGC-27 and SGC-7901 were treated with white lentil polysaccharides,the mitochondrial membrane potential of the cells was significantly reduced,proliferation activities of the cells were inhibited obviously( P 0.01),and the inhibition degree was related with drug concentration and incubation time. Flow cytometry analysis showed that the apoptosis rates of HGC-27 and SGC-7901 cell lines were 53. 15% and 38. 77% respectively,which were significantly increased compared with PBS treatment groups( 8. 07% and 6. 03%,P 0. 01). And the cell cycle of these cells was not influenced significantly. The results of QPCR showed that Bcl-2 gene transcription level of HGC-27 and SGC-7901 cell lines was significantly down-regulated,and the transcription levels of Bax and caspase-3 genes were significantly increased( P 0. 01). Conclusion: The white lentil polysaccharides can induce the apoptosis of gastric cancer cells lines HGC-27 and SGC-7901,and the mechanism of their apoptosis may be mediated by the regulation of Bax-Bcl-2-caspase3 pathway.
作者
张艳姿
柯瑞君
蒋盼若
杨林军
陈佳玉
ZHANG Yan-zi;KE Rui-jun;JIANG Pan-ruo;YANG Lin-jun;CHEN Jia-yu(School of Laboratory Medicine and Life Sciences,Wenzhou Medical University,Wenzhou 325035;Medical School,Shaoxing University,Shaoxing 312000;Taizhou Municipal Hospital,Taizhou 318000;Taizhou University Medical School,Taizhou 318000,China)
出处
《中国应用生理学杂志》
CAS
CSCD
北大核心
2018年第3期268-272,共5页
Chinese Journal of Applied Physiology
基金
国家自然科学基金资助项目(81402979)
浙江省科技厅资助项目(2015C33270)
浙江省卫生厅资助项目(2014KYA230)