Experimental Study On The Effect Of Shenqi Fuzheng Injection Combined With Interferon Alpha On Human Liver Cancer Cells

Objective:To Investigate the affect of MHCC97L hepatoma cell proliferation and apo ptosis after Shenqifuzheng Injection combining with Interferon-a acting on STAT1 silen cing to further clarify the mechanism of Shenqifuzheng Injection combining with Interf eron-a inhibiting liver cancer recurrence and metastasis.Methods:The experiments were divided into three groups,that is Blank control group(MHCC97L cells were added to DMEM with 10%FBS culture medium),STAT1&R NAi group(That is to add pLKO-1-STAT1-1 plasmid when transfection),LV&RNAi group(That is to add empty vector Lentiviral Particles when transfection).Cells in e ach group were used in two different ways to culture,that is the dose of 6000IU of re combinant human interferon-alb injection combined with a concentration of 0.5g/L S henqifuzheng injection(Hereinafter referred to as the combined group)or to join the bl ank media.To use the method of cell proliferation assay(MTT),vitro invasion assay(Tra nswell),researching the effects of senqifuzheng injection in combination with IFN-? on cell proliferation and cell invasion of MHCC97L STAT1 gene after transfection of R NAi and using western Blot change/RT-PCR technology to detect the changes of mRN A and protein of STAT1.Results:When the cell concentration of 4×104/ml,a dose of 6000IU interferon in com bination with a small dose of Shenqifuzheng injection acting 72 hours of STATIRNAi-transfected MHCC97L hepatoma cell is lower in MTT than a dose of interferon in c ombination with a small dose of 6000IU Shenqifuzheng injection acting 72 hours on t he inhibition of human hepatoma MHCC97L.Significant difference was statistically sign ificant(P<0.001).RT-PCR results showed that after the combined effects of the two dr ugs in hepatoma MHCC97L cells for 16h,its STAT1mRNA expression was significantl y up-regulated,nine times higher than the expression level of the control group(P<0.05).Joint group acting on MHCC97L cells before and after the STAT1 gene silencing,t he expression of its STAT1mRNA non-transfected cells were significantly higher than t ransfected group(P<0.05).Significant difference was statistically significant.Former ST AT1mRNA expression of transfected medication compared with untreated group,the di fference was not statistically significant.Untransfected group STAT1mRNA expression tr eatment group was significantly higher than the untreated group,the difference was sta tistically significant.Western Blot results showed that after Shenqifuzheng injection co mbined with recombinant human interferon a-lb injection acting on hepatoma cell MH CC97L for 16h,STAT1 protein expression was significantly higher than the control gr oup(P<0.01).No significant difference before and after treatment were transfected to e xpress the amount of STAT1 protein.No expression of STAT1 protein transfection gro up was significantly higher than the transfected cells after the combination therapy.Conclusion:Small doses of Shenqifuzheng injection(equivalent adult 100mg/d dosage)collaborative recombinant human interferon ?-1b injection can enhance liver MHCC97-L cell line.Its specific mechanism for the Shenqifuzheng injection combined with inter eron a can be activated JAK/STATs signaling pathway enhanced expression of STAT1 to inhibit the proliferation of liver cancer cells and promote apoptosis of hepatoma cel ls.Shenqifuzheng injection by the action of JAK/STATs signaling pathway reduce resist ance acting on the liver to produce interferon-?,but whether it is the only path require s further experiments confirmed.