摘要
目的:探讨二氢杨梅素对人类肝癌细胞株HepG2的抑制生长和诱导凋亡作用及其机制。方法:采用四甲基噻唑蓝比色法测试二氢杨梅素对细胞死亡率和细胞增殖活力的影响,并计算半抑制浓度IC50;通过HE荧光染色法和Annexin V-PI流式细胞术分别观察和分析二氢杨梅素对肝癌细胞HepG2各个细胞周期的影响,最后利用Western印迹法对与细胞周期和凋亡相关的蛋白群p34cdc-2、CyclinB1、Bcl-2和PARP在二氢杨梅素作用前后的表达量进行研究。结果:肝癌细胞HepG2的细胞死亡率随着二氢杨梅素溶液浓度的升高而升高,IC50值为(35.22±1.56)μmol/L,且呈现良好量效关系;二氢杨梅素对肝癌细胞HepG2各个细胞周期均有显著影响,其中G2/M期细胞比率剧增、S期细胞比率大幅降低,而G0/G1期细胞比率则呈指数级减少,与阴性对照组比较,差异具统计学意义(P<0.05);p34cdc-2蛋白的表达量未见变化,CyclinB1蛋白的表达量剧增,Bcl-2蛋白的表达量大幅下降,而PARP蛋白则因发生水解而表达量有所减少。结论:二氢杨梅素对人类肝癌细胞株HepG2具显著的抑制生长和诱导凋亡的药理作用,其作用机制是通过线粒体信号转导途径,激活含半胱氨酸的天冬氨酸蛋白水解酶Caspase-3,进而参与细胞凋亡。
Objective To discuss the proliferation inhibition, apoptosis-inducing effects and the mechanism of dihydromyricetin on human liver cancer HepG2. Methods The effects of dihydromyricetin on cell death rate and proliferation in light of MTT test were examined and IC50 was calculated. The effects of dihydromyricetin on liver cancer HepG2 in different cell cycles were observed and analyzed in light of HE fluorescent staining method and Annexin V-PI flow cytometry （FCM）. The protein expressions of p34^-2,CyclinB1,Bcl-2 and PARP which were related to the cell cycle and apoptosis before and after the use of dihydromyricetin were determined in light of Western blotting. Results The death rate of liver cancer HepG2 were increased with the increasing of dihydromyricetin concentration. The IC50 read （35.22 ± 1.56）μmol/L and displayed dose-response relationship. Dihydromyricetin had significant impacts on liver cancer HepG2 in different cell cycles, the proportion of G2/M cells rose dramatically and that of S cells lowered sharply and that of G0/G1 cell decreased exponentially.The difference was of statistical significance compared with the control group （P 〈 0.05）. Conclusion Dihydromyricetin has distinct medical effects on inhibiting the growth and inducing the apoptosis of liver cancer HepG2. The mechanism is to activate Caspase-3, thus engaged in apoptosis in light of signal transduction of the mitochondrion.
出处
《实用医学杂志》
CAS
北大核心
2014年第11期1712-1715,共4页
The Journal of Practical Medicine
