华蟾素注射液对人肝癌肿瘤细胞HepG_2增殖抑制作用观察及其作用机制研究

Inhibitory Effects Observation on Cinobufagin Injection on Proliferation of Human Liver Cancer Cells HepG_2 and its Mechanism Investigation

目的探讨华蟾素注射液对人肝癌肿瘤细胞Hep G2增殖抑制作用观察及其作用机制研究。方法 MTT法测定华蟾素注射液对Hep G2细胞增殖抑制作用,高内涵活细胞成像系统检测华蟾素注射液对Hep G2细胞周期和细胞凋亡的影响,Western blot蛋白印迹法检测华蟾素注射液凋亡相关蛋白的表达。结果华蟾素注射液不同浓度作用于24h和48h抑制率均明显高于阴性对照组(P<0.05),且随着浓度的增加,抑制率越为明显;华蟾素注射液不同浓度作用于48h抑制率明显高于24h抑制率(P<0.05);华蟾素注射液不同浓度G0/G1期百分率高于阴性对照组(P<0.05),S期细胞百分率低于阴性对照组(P<0.05);华蟾素注射液不同浓度凋亡率高于阴性对照组(P<0.05);随着华蟾素注射液浓度增加细胞凋亡率增加;随着华蟾素注射液浓度的减少,Bcl-2蛋白表达不断增加,而Bax蛋白表达不断下降。结论华蟾素注射液可明显抑制人肝癌肿瘤细胞Hep G2增殖,阻滞细胞于G0/G1期,可能通过上调bax蛋白表达、下调Bcl-2蛋白表达相关。

Objective To investigate the inhibitory effects of Cinobufagin injection on the proliferation of human liver cancer cells HepG2 and its mechanisms of action. Methods MTF assay was used to determine the proliferating of HepG2 cells the of high content live cell imaging systems to was used detect the inhibitory effects of Cinobufagin injection on cell cycle and apoptosis in HepG2cells. Western blot method was used to detect the expression of Cinobufagin injection of apoptosis related protein. Results 24h and 48h inhibition rates of different concentrations of Cinobufagin injection were significantly higher than those in negative control group （P 〈 0.05 ）, with the concentration increased, the inhibition rate was more obvious; the inhibitory rates of 48h of different concentrations of Cinabufagin injection was significantly higher than those of 24h inhibition rates （ P 〈 0.05 ） ; different concentrations of G0/G1 phase percentage of Cinobufagin injection higher than those of negative control group （ P 〈 0.05）, the percentage of S phase cells was lower than that of negative control group （ P 〈0. 05 ） ; the apoptosis rate of different concentrations of Cinobufacini injection was higher than those of negative control group （ P 〈 0.05 ） ; with the increase of the concentration of Cinobufacini injection, the rate of apoptosis increased; with the decrease of cinobufacini injection concentration, bcl -2 protein expression increased, Bax protein expression decreased. Conclusion Cinobufagin injection can obviously inhibit the proliferation of human liver cancer HepG2tumor cells and arrest cells in G0/G1 phase, it may be related to the up - regulation of Bax protein expression and down - regulation of Bcl - 2 protein expression.