青藤碱抗肝癌作用机制研究  

Study on anti-tumor mechanism of sinomenine

作　　者：王高峰[1] 张强[1] 张利娟[1] WANG Gao-Feng ZHANG Qiang ZHANG Li-Juan.(Experimental Center of Medical College, Huanghe University of Science and Technology, Zhengzhou 450063, Chin)

机构地区：[1]黄河科技学院医学院实验中心,郑州450063

出　　处：《中国免疫学杂志》2017年第5期688-692,共5页Chinese Journal of Immunology

摘　　要：目的:探讨青藤碱对Hep G2人肝癌细胞株的增生和转移能力的多靶向作用机制。方法:运用噻唑蓝还原法(MTT)来检测青藤碱对Hep G2人肝癌细胞株的抗增殖作用,而同时运用Transwell法来检测药物对于细胞转移能力的作用变化;间接荧光标记法测定经过不同浓度的青藤碱作用后,细胞内活性氧水平的变化;利用酶抑制动力学方法,研究青藤碱对逆转录酶的抑制作用;再通过逆转录聚合酶链式反应和蛋白质印迹的实验来检测Hep G2细胞中凋亡蛋白水平的变化。结果:青藤碱对Hep G2人肝癌细胞株具有抗侵袭及转移的治疗作用。MTT检测结果显示青藤碱对于Hep G2人肝癌细胞抗增殖作用明显,半抑制浓度IC50为(15.35±2.43)μmol/L;而Transwell法的结果显示青藤碱对癌细胞有较好的抗转移能力;青藤碱是一种有效的逆转录酶抑制剂,IC50为(21.32±2.43)μmol/L;荧光标记法检测细胞内活性氧水平随青藤碱呈浓度依赖性升高;蛋白水平测定显示青藤碱上调Hep G2人肝癌细胞CASP3、CASP9、CAV1和下调SOX2的表达。结论:青藤碱可能是通过上调CASP3、CASP9、CAV1和下调SOX2的表达,进而抑制人肝癌细胞的增殖和转移,调节相关信号通路,最终在分子水平证明青藤碱对Hep G2人肝癌细胞的抑制作用。Objective：To investigate the anti-proliferation and anti-metastasis effects and study the molecular mechanism of si-nomenine in cell line（HepG2）.Methods： HepG2 cells were cultured together with different treatment concentrations of sinomenine.The effect of sinomenine on inhibition of growth of HepG2 cells were determined by methyl thiazolyl tetrazolium（MTT） assay.The effect of sinomenine on inhibiting metastasis of HepG2 cells were determined by Transwell assay.The inhibitory effect of sinomenine on reverse transcriptase（RT） was studied using inhibitory kinetic method,on the basis,the reactive oxygen species（ROS） of HepG2 cells was monitored by indirect fluorescent labeling.The protein expressions of CASP3,CASP9,CAV1 and SOX2 were analyzed by Western blot experiment.Results： Sinomenine inhibited the proliferation and metastasis of HepG2 cells significantly.Sinomenine had a good inhibitory effect on the growth of HepG2 cells,half inhibitory concentration（IC50） was （15.35±2.43） mol/ L.Sinomenine was RT inhibitor,IC50 was （21.32±2.43） mol/ L.The Western blot showed that CASP3,CASP9 and CAV1 were up-regulated and SOX2 was down-regulated by the sinomenine treatment in HepG2 cells.Conclusion： The potential molecular mechanism of sinomenine suppresses proliferation and metastasis of HepG2 cells by up-regulation of CASP3,CASP9,CAV1 and down-regulation of SOX2.

关 键 词：青藤碱 HEPG2人肝癌细胞 活性氧水平 逆转录酶 CASP3 CASP9 CAV1 SOX2 

分 类 号：R453.9[医药卫生—治疗学]