Oxymatrine Effects The Expression Of ARID1A In Hepatocellular Carcinoma Cell Lines MHCC97L And MHCC97H

Objective To test changes of ARID1 A expressions in cell lines MHCC97 L,MHCC97H and L02,and to observe the effect of oxymarine(OM)on inhibited cell proliferation in different metastic potential hepatocellular carcinoma cell lines(MHCC97L and MHCC97H)and its effect on the expression of ARID1 A.To study the possible mechanism of OM on inhibiting the hepatocellular carcinoma.Methods The MHCC97 L,MHCC97H and normal human L02 cell lines were used in this study.Firstly,the cell immunofluorescence microscopy was applied to localize ARID1 A expression in three cell lines.Secondly,the mRNA expression of ARID1 A was tested in these cell lines by qRT-PCR,and Western blot was used to analyze the expression of BAF250 a protein in these cell lines.Thirdly,MTT was used to measure the impact of OM of different concentrations on the proliferation of MHCC97 L and MHCC97 H cells.Finally,the mRNA and BAF250 a protein expressions of ARID1 A were tested after the OM intervention by qRT-PCR and Western blot in the MHCC97 L and MHCC97 H cells.Result(1)The immunofluorescence microscopy showed that the ARID1 A protein located in the nucleus in the MHCC97 L,MHCC97H and L02 cells.(2)qRT-PCR analyses showed the mRNA expressions of ARID1 A in the MHCC97 L and MHCC97 H cells were lower than those in the L02 cells(P<0.05).However,the relative quantitative mRNA expressions of ARID1 A in the MHCC97 L and MHCC97 H cells had no significant difference.(3)Western blot showed that the expressions of BAF250 a protein in the MHCC97 L and MHCC97 H cells were lower compared with the L02 cells(P<0.05).But when the expressions of BAF250 a protein in the MHCC97 L cells were compared with those in the MHCC97 H cells,the difference was insignificant(P>0.05).(4)The result from MTT showed that,after the OM intervention in MHCC97 L and MHCC97 H cell lines,the effect of OM on cell proliferation inhibition was in a dose-dependent manner,and with the increasing of concentration,the cell proliferation inhibition was increased(P<0.05).Nevertheless,the effect of OM on MHCC97 L and MHCC97 H cell proliferation inhibition was not in a time-dependent manner(P > 0.05).(5)qRT-PCR analyses showed the mRNA expressions of ARID1 A,after the OM intervention,in the groups(0.5mg/ml~2.0mg/ml)in the MHCC97 L and MHCC97 H cell lines were lower than the groups(0mg/ml),but the relative quantitative expressions were on the rise in a concentration(1.0mg/ml~4.0mg/ml)(P < 0.05).Furthermore,the mRNA expressions of ARID1 A in the MHCC97 L and MHCC97 H cells had no difference in a same concentration(P>0.05).(6)Western blot showed that,after the OM treatment,the expressions of BAF250 a protein in the MHCC97 L and MHCC97 H cell lines were higher in the concentration of 0.5mg/ml~4.0mg/ml than in the concentration of 0mg/ml(P<0.05).Moreover,the expressions of BAF250 a protein in the MHCC97 L cells were higher than those in the MHCC97 H cells in same concentration of 0.5 and 1.0mg/ml(P>0.05).Conclusion There were different expressions in hepatocellular carcinoma cell(HCC)lines and in the L02 cells.The expressions of ARID1 A were lower in the MHCC97 L and MHCC97 H cells than those in the L02 cells.Oxymatrine might have inhibitory effects on MHCC97 L and MHCC97 H cells,and could enhance the protein expression of ARID1 A in MHCC97 L and MHCC97 H cell lines.The effects were more significant in low-grade malignant MHCC97 L cells.These results indicated that enhancement of the protein expression of ARID1 A may be one of the mechanisms of the inhibitive effect of OM on hepatocellular carcinoma cell proliferation.