中药单体蟾毒灵对人肝癌细胞HepG2的抑制作用及其机制研究
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摘要
目的探讨蟾毒灵抑制人肝癌细胞株HepG2增殖的作用及其相关机制。方法MTT方法检测蟾毒灵对肝癌细胞HepG2生长抑制作用;FCM方法检测蟾毒灵单体诱导绌胞细胞周期周期阻滞及凋亡的作用;Western blot方法观察蟾毒灵单体对AKT以及JNKMAPK信号通路的作用、对细胞周期相关蛋白及凋亡相关蛋白的影响。结果(1)蟾毒灵能够抑制HepG2细胞的生长,且抑制作用呈时间依赖性和剂量依赖性。(2)蟾毒灵能够下调cyclinB1的表达,使MPF失活,从而使HepG2细胞周期阻滞于G2/M期。(3)蟾毒灵单体能够下调Bcl-2的mRNA水平和上调Bax的mRNA水平,从而使Bcl-2蛋白表达下降、Bax蛋白表达升高,从而诱导HepG2细胞的凋亡。(4)上述蟾毒灵导致的细胞周期阻滞与诱导凋亡现象可能与蟾毒灵抑制AKT信号通路和活化JNK MAPK信号通路有关。
结论蟾毒灵可以改变某些细胞信号通路,诱导细胞周期阻滞和细胞凋亡从而抑制肝癌细胞生长。
Objective To study the antiproliferative effects of bufalin on hepatocellular carcinoma cell line HepG2 and the relative mechanisms. Methods To study the inhibitive effects of bufalin on the proliferation of HepG2 cells by MTT assay, apoptosis and cell cycle distribution by flow FCM. Western blotting is used to detected the Expression of protein related to cell cycle and apotosis and the activity of survival signal transduction pathway such as PKB/AKT and JNKMAPK.Results (1)Bufalin could inhibit the growth of HepG2 cell in a concentration and time dependent manner in vitro. (2) Bufalin could cause G2/M cell cycle arrest in HepG2 cells through downregulating the leval of CyclinB1 and inactivating CDK1.(3) Bufalin could induce remarkable apoptosis in HepG2 cells, the apoptosis effect was partly caused by the up-regulated leval of mRNA and protein of Bax accompanied with down-regulated leval of mRNA and protein of Bcl-2.(4) The above cell cycle arrest and apoptosis was related to the activation of JNK MAPK signal pathway and inhibition of PKB/AKT signalpathway.
结论蟾毒灵可以改变某些细胞信号通路,诱导细胞周期阻滞和细胞凋亡从而抑制肝癌细胞生长。
Objective To study the antiproliferative effects of bufalin on hepatocellular carcinoma cell line HepG2 and the relative mechanisms. Methods To study the inhibitive effects of bufalin on the proliferation of HepG2 cells by MTT assay, apoptosis and cell cycle distribution by flow FCM. Western blotting is used to detected the Expression of protein related to cell cycle and apotosis and the activity of survival signal transduction pathway such as PKB/AKT and JNKMAPK.Results (1)Bufalin could inhibit the growth of HepG2 cell in a concentration and time dependent manner in vitro. (2) Bufalin could cause G2/M cell cycle arrest in HepG2 cells through downregulating the leval of CyclinB1 and inactivating CDK1.(3) Bufalin could induce remarkable apoptosis in HepG2 cells, the apoptosis effect was partly caused by the up-regulated leval of mRNA and protein of Bax accompanied with down-regulated leval of mRNA and protein of Bcl-2.(4) The above cell cycle arrest and apoptosis was related to the activation of JNK MAPK signal pathway and inhibition of PKB/AKT signalpathway.
引文
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