Effects Of Matrine On Migration And Invasion Of Hepatocellular Carcinoma HepG2 Cells By Regulating MiR-122 And Its Molecular Mechanism

Objective: To explore the role of matrine in regulating the invasion and metastasis of HepG2 cells through miR-122 and its molecular mechanism.Methods: The transient transfection method was used to transfect HepG2 cells with miR-122 to low expression of miR-122,and the transfection efficiency was observed under fluorescence microscope;CCK-8 was used to detect the effect of matrine with concentrations of 0.5mg/ml,1mg/ml,2mg/ml,4mg/ml and 8mg/ml on the survival rate of HepG2 cells;The effect of matrine on the invasion and metastasis of hepatocellular carcinoma was detected by cell scratch and Transwell method;The expression of miR-122 was detected by q PCR after matrine acted on HepG2 cells,and The expression of EMT-related proteins E-cadherin,N-cadherin and Vimentin were detected by Western blot.And the expression of key factors RhoA,Rock1 and Rock2 in RhoA/Rock signaling pathway.Results:(1)Matrine could significantly inhibit the cell activity of HepG2 cells after 48 hours of treatment.The cell activity of matrine at the concentrations of 0.5mg/ml,1mg/ml,2mg/ml,4 mg/ml and 8 mg/ml were(75.34±9.84),(57.60±7.95),(36.09±3.21),(27.77±10.96)and(23.77±10.14),respectively(P<0.001);(2)Matrine can inhibit the migration and invasion of cells.The scratch healing rate of the miR-122mimic+ Matrine group,the Matrine group and the miR-122 mimic group were(8.14±0.43),(21.99±1.53)and(16.02±2.64),respectively,which were lower than those of the negative control group(29.80±1.05)(P<0.001);(3)The number of penetrating cells of the miR-122 mimic+ Matrine group,the Matrine group and the miR-122 mimic group were(11.00±3.61),(35.67±8.33)and(43.33±12.22),respectively,They were all less than(106.3±9.29)in the negative control group(P<0.001);(3)Matrine and overexpression of miR-122 can down-regulate the expression of migration marker proteins RhoA,ROCK1 and ROCK2 in HepG2 cells.The relative expression levels of RhoA in miR-122 mimic+Matrine group,Matrine group and miR-122 mimic group were(48.89±13.21),(67.05±9.72),and(69.21±11.11),higher than those of negative control group(100.01±12.17)(P < 0.004),the relative expression level of ROCK1(45.18±10.5),(65.87±6.08),(83.08±4.86)were lower than those of negative control group(101.09±5.83)(P < 0.001).The relative expression level of ROCK2 was(28.19±10.28),(71.23±9.16)and(78.86±6.30),respectively,which were lower than that of negative control group(110.0±18.24)(P < 0.001);(4)Matrine and overexpression of miR-122 could up-regulate the expression of E-cadherin,the migration marker protein of HepG2 cells,and down-regulate the expression of Ncadherin and Vimentin.The relative expression of E-cadherin in miR-122mimic+Matrine group,Matrine group and miR-122 mimic group were(94.77±8.01),(81.83±12.76),(74.82±9.53),respectively,which were higher than those in the negative control group(41.22±9.06)(P<0.001),and the relative expression of Ncadherin was(45.36±8.73),(55.86±3.53)(75.55±7.86)were lower than the relative expression of the negative control group(107.07±16.66)(P<0.001),and the relative expression of Vimentin was(65.23± 9.80),(82.46±5.03),and(84.73±5.91)were lower than the relative expression of the negative control group(113.84±9.44)(P<0.001).Conclusion:(1)Matrine can inhibit the proliferation of hepatoma HepG2 cells;(2)Matrine inhibited the migration and invasion of hepatoma HepG2 cells by upregulating the expression of miR-122.(3)Matrine regulates the expression of miR-122 to inhibit EMT process and RhoA/ROCK signaling pathway activity,and then regulates the migration and invasion of hepatoma HepG2 cells.