摘要
以茶氯香酰胺脂质体(TClC-L)为研究对象,研究其对人肝癌细胞(SMMC7721)生长和迁移的影响,并且探索其作用的分子机制,为研发抗肝癌新药提供科学依据.应用MTT法检测TClC-L对SMMC7721细胞生长的影响;应用划痕法和细胞流式分别检测TClC-L对SMMC7721细胞迁移和死亡的影响;应用Hoechst 33258染色检测SMMC7721细胞凋亡形态学变化;应用Western blotting技术检测TClC-L对SMMC7721细胞生长、凋亡和迁移密切相关蛋白表达的影响和其可能作用的分子机制.结果显示,TClC-L对SMMC7721细胞生长有显著的抑制作用,显示出剂量和时间的依赖性关系; TClC-L能够显著地降低SMMC7721细胞的迁移能力,促进其凋亡; TClC-L显著上调抑制肝癌细胞生长和诱导癌细胞凋亡的相关蛋白因子Bax、Caspase-3、PARP和迁移抑制因子E-cadherin蛋白的表达水平,下调促进细胞生长和迁移以及抗凋亡的相关蛋白因子Bcl-2、HGF、cMet、NF-κB、MMP9、VEGFR2、c-Myc蛋白的表达水平. TClC-L抑制人肝癌细胞SMMC7721的生长和迁移作用的分子机制涉及到HGF/c-Met/VEGFR-NF-κB信号通路.
In this study, the inhibitory effects of TClC-L on the growth and migration of human hepatocellular carcinoma cells (SMMC7721) are studied, and its molecular mechanism of action is explored to provide a scientific basis for the future development of a new anti-hepatocellular carcinoma drug. The MTT assay is used to detect the effect of TClC-L on the growth of SMMC7721 cells. The scratch method and flow cytometry are applied to evaluate the effects of TClC-L on the migration and cell death of SMMC7721 cells, respectively;The morphological changes of apoptotic SMMC7721 cells are detected by Hoechst 33258 staining. The Western blotting is used to explore the effect of TClC-L on the expressions of cell growth, apoptosis and migration-related proteins and its possible molecular mechanism of action. The results show that TClC-L significantly inhibits the growth of SMMC7721 cells in a dose-and time-dependent manner. TClC-L significantly reduces the migration ability of SMMC7721 cells and promotes the apoptosis of SMMC7721 cells. TClC-L markedly up-regulates the expressions of Bax, caspase-3, PARP and E-cadherin proteins related to the inhibition of growth and migration of hepatoma cells and induced the apoptosis in hepatoma SMMC7721 cells. TClC-L also remarkably down-regulates the protein expressions of Bcl-2, HGF, c-Met, NF-κB, MMP9, VEGFR2 and c-Myc, which promote cell growth and migration, and anti-apoptosis. The molecular mechanism of TClC-L action for inhibiting the growth and migration of human hepatocellular carcinoma SMMC7721 cells is related to the suppression of HGF/c-Met/VEGFR-NF-κB signaling pathway.
作者
张伟伟
张欣
石晓玉
张燕
王朔
刘昆
张国营
ZHANG Wei-wei;ZHANG Xin;SHI Xiao-yu;ZHANG Yan;WANG Shuo;LIU Kun;ZHANG Guo-ying(Laboratory of Molecular Pharmacology, School of Pharmacy, Yantai University, Collaborative Innovation Center of Advanced Drug Delivery System and Biotech Drugs in Universities of Shandong, Key Laboratory of Molecular Pharmacology and Drug Evaluation,(Yantai University) Ministry of Education, Yantai University,Yantai 264005,China)
出处
《烟台大学学报（自然科学与工程版）》
CAS
2019年第2期151-158,共8页
Journal of Yantai University(Natural Science and Engineering Edition)
基金
山东省自然科学基金资助项目(ZR2015HM004)
