Influence Of Oxymatrine On Proliferation Of L-02and HepG2and Expression Of MircroRNA-122and MircroRNA-21in HepG2

Objective: In order to provide the experimental basis for mechanism of theoxymatrine(OM) anti-hepatOMa that we investigate the effect of oxymatrine on theproliferation of human liver cells L-02and human hepatocelluar carcinOMa cellline HepG2, and the expression of MicroRNA-122and MicroRNA-21wereinvestigated in the HepG2.Methods: Human liver cells L-02and Human hepatocelluar carcinOMa cellline HepG2were cultured in vitro. With no oxymatrine as negative control group,different dose of OM as the experimental group, the influence of OM on theproliferation of L-02and HepG2were examined by the method of MTT. Whendifferent dose of OM(1/3IC50,2/3IC50, IC50) were added and maintained for48hours in HepG2,which the cell cycle and apoptosis were detected by using FlowcytOMetry. The expression of MicroRNA-122and MicroRNA-21in humanhepatocelluar carcinOMa cell line HepG2,which were treated with IC50oxymatrinefor72h, then were detected by real-time PCR assay.Results: The MTT assay showed that：1.The effect of oxymatrine on human liver cells L-02proliferation:(1) When the role of time in the24h, cOMpared withthe negative control group, there is no proliferation inhibition in experimentalgroups with the concentration oxymatrine of0.5mg/ml,1.0mg/ml,2.0mg/ml,respectively (P＞0.05), when OM concentration increased to3.0mg/ml,4.0mg/ml,8.0mg/ml,there are different degrees of inhibition for L-02proliferation (P <0.05),the inhibition rate was8.48%,15.43%,51.72%,,respectively.(2) The inhibition rateof oxymatrine with the concentration of1.0mg/ml、2.0mg/ml、3.0mg/ml、4.0mg/mland8.0mg/ml on human liver cells L-02for48h and72h was18.79%,26.06%,27.58%,31.52%,76.36%and33.20%,38.53%,41.82%,55.61%,89.95%,respectively. COMpared with24h,48h and72h on human liver cells L-02proliferation has different degrees of inhibition (P <0.05or P <0.01).2. Theinhibition rate of oxymatrine on human hepatOMa cell line HepG2proliferationwith the concentration of0.5mg/ml、1.0mg/ml、2.0mg/ml、4.0mg/ml and8.0mg/mlfor48h and72h was6.93%、16.95%、33.74%、53.78%、65.95%�'�9.40%、20.58%、38.82%、57.80%、86.56%, respectively. COMpared with negative control group,there is significant difference in inhibited effect of oxymatrine on the proliferationof human hepatocelluar carcinOMa cell line HepG2, respectively (P <0.05), andwith the increase of oxymatrine concentration and prolonging the time of action,the inhibition effect of oxymatrine on the human hepatocellular carcinOMa cell lineHepG2was more obvious, with time-and dose-dependent. Flow cytOMetryanalysis showed that: cOMpared with the negative control group, with theconcentration of1/3IC50,2/3IC50, IC50on the HepG2for48h, G0/G1phasecells increased gradually, and G2phase cells decreased gradually; the apoptotic ratewas12.5%,15.6%and23.3%, respectively. COMpared with the control group, there is statistically significant, respectively (P <0.05). The result of Real-TimePCR showed that MicroRNA-122was up-regulated and MicroRNA-21wasdown-regulated,which were treated by the IC50oxymatrine, and their up-downratio were2.79times and0.44times, respectively.Conclusions:1. There have no significant injury or toxic side effects onhuman liver cells L02when OM there is at low concentrations within a short periodof time, there have different degrees of injury or toxic side effects on human livercells L-02when with the increase of OM concentration and the action of timeprolongation, and there was time and dose dependent.2. Oxymatrine would haveobvious inhibition on proliferation in human hepatocelluar carcinOMa cell lineHepG2, and there was time and dose dependent. The mechanism of theoxymatrine anti-hepatOMa may be blocked the cell in G0/G1phase, inhibited theproliferation, and prOMoted the apoptosis.;and yet may be concerned to theup-regulation of MicroRNA-122and down-regulation of MicroRNA-21.