摘要
目的探讨一种新齐墩果酸衍生物(3β-hydroxyolea-12-en-28-oic acid-3,5,6-trimethylpyrazin-2-methyl ester,简称为受试物)对人肝细胞癌细胞株SMMC-7721细胞的抗肿瘤作用机制。方法应用四甲基偶氮唑蓝(MTT)法及细胞形态学观察新齐墩果酸衍生物对SMMC-7721细胞增殖及形态的影响;Annexin V联合PI染色法检测新齐墩果酸衍生物对细胞膜以及用JC-1染色法检测线粒体膜电位的损伤程度;Western blotting检测新齐墩果酸衍生物对凋亡相关蛋白Bcl-2,Bax,caspase-9 and caspase-3的表达。结果新齐墩果酸衍生物能抑制人SMMC-7721细胞增殖力,与浓度和时间呈正相关,作用细胞48 h后,细胞出现核固缩,细胞裂解,产生凋亡小体等形态学改变,细胞膜完整性被破坏,线粒体膜电位(Δψm)被降低,Bax、pro-caspase-9、cleaved caspase-9、pro-caspase-3以及cleaved caspase-3蛋白表达水平被上调,Bcl-2蛋白表达水平被下调。结论这种新齐墩果酸衍生物在人SMMC-7721细胞中发挥抗癌作用可能与其破坏了线粒体功能有关。
OBJECTIVE To investigate the anti-tumor mechanisms of a new oleanolic acid derivative （the test substance, 3β- hydroxyolea-12-en-28-oic acid-3,5,6-trimethylpyrazin-2-methyl ester） on the human hepatocellular carcinoma SMMC-7721 cells. METHODS MqT assay and morphology observation were used to determine the effects of the new oleanolic acid derivative on human hepatocellular carcinoma SMMC-7721 cells. Cell membrane integrity was assessed with AV/PI uptake by fluorescence microscope. The loss of mitochondrial membrane potential （AOm） was detected by using JC-1 assay. Western blotting was used to detect the apoptosis- related proteins Bcl-2, Bax, caspase-9 and caspase-3. RESULTS The new oleanolic acid derivative reduced the cell viability of SMMC-7721 cells in dose- and time-dependent manner. After treatment of the new oleanolic acid derivative on cells for 48 h, cell nucle- us pycnosis, cell lysis and the generation apoptosis body and so on were observed, the cell membrane integrity was damaged, the mito- chondrial membrane potential （A^m） was decreased, the protein levels of Bax, pro-caspase-9, cleaved caspase-9, pro-caspase-3 and cleaved caspase-3 were upregulated, and the protein level of Bcl-2 was downregulated. CONCLUSION The growth inhibition and pro- apoptotic effects of this new oleanolic acid derivative on human SMMC-7721 cells associate with its damage on mitochondrial function.
作者
范新凤
王鹏龙
杜海渊
孙耀贵
段智变
雷海民
李宏全
FAN Xin-feng WANG Peng-long DU Hai-yuan SUN Yao-gui DUAN Zhi-bian LEI Hai-min LI Hong- quan(College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, China College of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing 100029, China)
出处
《中国药学杂志》
CAS
CSCD
北大核心
2016年第24期2114-2118,共5页
Chinese Pharmaceutical Journal
基金
国家农业科技成果转化资金项目(2014GB2A300003)
山西农业大学科技创新基金(J141602002)
