Mechanistic Study Of Oxymatrine’ Invasionand Metastasis Effects On Hepatocellular Carcinoma Cell Line HepG2

Objective To investigate the influence and the possible molecular mechanisms of oxymatrine (OM) affect the invasion and metastasis of hepatocellular carcinoma cell line HepG2, and to detect the effect of expression of E-cadherin and CD44, MMP2 and TIMP2, to provide the basis of applying the oxymatrine at anti-hepatoma.Methods The human hepatocellular carcinoma cell line HepG2 was used in this study. The cell line HepG2 was divided into three groups:A blank control group, a negative control group and OM group. Besides,the OM group was divided into 5 groups(0.5mg/ml, 1.0mg/ml,2.0mg/ml,4.0mg/ml and 8.0mg/ml). Measuring the effect of OM on HepG2 cell proliferation by the method of MTT. Detect the invasion and migration ability of HepG2 cell was through the cell scratch test and Transwell Chambers method,and the expression of E-cadherin、CD44、MMP2 and TIMP2 on OM group (a 1.0 mg/ml group、a 2.0 mg/ml group and a 4 mg/ml group) were tested by Real-time PCR and Western Blot.Results The data from MTT indicated that with the longer effect-time and drug concentration, the affect of OM on inhibiting HepG2 cell proliferating was depended on its concentration and effect-time; the concentration of 1.0mg/ml, 2.0mg/ml and 4.0mg/ml was selected as the final experimental group.the informations from Scratch experiment about each OM group manifested that the higher the drug concentration was,the slower the scratches wound healed.Transwell trial showed that compared with the negative control group, the invasion and metastasis ability of each OM group was significantly decreased (P< 0.05).what’s more, it depended on the concentration gradient.The results from RT-PCR and Western Blot suggested that the expression of mRNA and protein on CD44 in OM groups were higher than control group,while the E-cadherin was lower.Real-time PCR and West Blot detection indicated that the E-cadherin and TIMP2 expression of mRNA and protein on the OM group were increased, while the CD44 and MMP2 were decreased.Conclusion The inhibitory effect of oxymatrine on the invasion and migration of HepG2 may be related to the expression of CD44, MMP2, TIMP2 and E-cadherin.