Research On Hepatic Carcinoma Cell Proliferation By Erk Signaling Pathway Via MIF

Objective: From the cellular level to research the molecular mechanism of hepatoma cells proliferation through ERK signaling pathway via MIF, for the further development in the treatment of HCC as a target of MIF to establish the theoretical basis.Methods: First, MIF(50 ng/ml, 100 ng/ml, 200 ng/ml) was added into HepG2 cells as the experimental group, cirrhosis QSG-7701 cells and normal liver cells L-02 as control groups, the MIF antagonist ISO-1(50 μM) added into HepG2 cells as a negative control group, also set up a control group in each group, only added the medium. HepG2 cell viability was detected by MTT assay to screen the optimum concentration of MIF. Secondly, cultured cells in each group for 24 h, ERK gene expression detected by RT-PCR and ERK 、p-ERK expression detected by Western-blot of each group.Results: 1.MIF obviously promote cell proliferation, the most significant role in HepG2;2.At the same time,cell proliferation were enhanced with the MIF concentration with MTT,the most significant concentration was 200 ng/ml,HepG2 was the most significant role; 3.To detect with Real time-PCR,ERK gene expression regulated, in which the highest MIF concentration(200 ng/ml) was significantly increased compared with the lower concentrations and the control group(P<0.05);The difference had statistically significant of ERK gene expression among HepG2 compared with QSG-7701 and L-02(P<0.05), but there is no difference between QSG-7701 and L-02(P>0.05). Western-blot results suggest that there is no significant difference in the expression of different concentrations of ERK protein expression, but p-ERK protein expression regulated(P<0.05), the p-ERK protein expression in MIF concentration(200ng/ml) was significantly increased compared with lower concentrations and the control group(P<0.05); The difference had statistically significant of p-ERK protein expression among HepG2 compared with QSG-7701 and L-02(P<0.05), but there is no difference between QSG-7701 and L-02(P>0.05). 4.ISO-1 inhibited HepG2 cell proliferation through decreasing the expression of ERK via inhibiting expression of MIF, to detect with Real time-PCR,ERK gene expression was lower compared with the MIFgroups,the difference was statistically significant(P<0.05).Western-blot results suggest that ERK protein expression was no significant difference in ISO-1 group compared with the MIF groups and control group, but the p-ERK protein expression decreased and has the statistically significant difference(P<0.05).Conclusion: MIF can promote HepG2 cells proliferation, also via the p-ERK signaling pathway to promote the hepatocellular carcinoma cells proliferation.