| Objective:To study the effect of NF-?B signaling pathway on the development of mouse HCC in mouse model of hepatocellular carcinoma(HCC)induced by AKT/c-Met co-expression,and to explore the role of NF-?B-related signaling pathway in HCC influences.The HCC model of AKT/c-Met mice was treated with tripterine to observe the effect of tripterine on the expression of NF-?B in mouse HCC model.Methods:1.AKT/c-Met co-expression promotes activation of NF-?B signaling pathwayThirty-two wild-type FVB/N mice were randomly divided into 4 groups,AKT group,c-Met group and AKT/c-Met group were injected with SB-mediated high-pressure tail vein transfection technique,WT group injection,etc.Volume of normal saline.After 6W modeling,blood was taken from the orbital venous plexus,and the serum levels of AST and ALT were determined.The mice were sacrificed under anesthesia,the liver tissues were dissected,photographed,and the records were weighed.A portion of mouse liver tissue was fixed with 4% paraformaldehyde.Tissue-embedded sections were stained with HE and observed under a microscope.Another part of the tissue was stored at 80 ° C and the expression levels of NF-?B and IL-6,TNF-?,COX-2 and PGE2 were detected by Western Bloting.2.Tripterine erythropoietin regulates NF-?B signaling pathway and induces AKT/c-Met co-expression to induce hepatocellular carcinoma in miceThirty-two wild-type FVB/N mice were randomly divided into 4 groups: normal group(WT),AKT/c-Met model group,tripterine low dose group(AKT/c-Met-Cel-1),tripterygium red High dose group(AKT/c-Met-Cel-2).Tripterine was administered for 4 weeks after modeling for 3 weeks(intraperitoneal administration,1 and 2 mg/kg/day).After the end of the administration,the mice were sacrificed under anesthesia,the liver tissues were peeled off,photographed,and the records were weighed.Western Bloting was used to detect the changes of PCNA and NF-?B and related protein levels in liver tissues of HCC mice after administration of tripterine.3.Effect of tripterine on NF-?B signaling pathway in HCC cell growth in vitroThe human hepatoma cells HepG2 and SMMC-7721 were cultured in H-DMEM medium containing 5% fetal bovine serum.When the cell density reached 80%,the cell protein was extracted and the expression level of NF-?B was detected.The results were compared.analysis.NF-?B-expressing cells were directly subjected to tripterine intervention,NF-?B-expressing cells were first transfected with AKT/c-Met plasmid,and then treated with tripterine;and different administration times and differences were investigated.The effect of the concentration of the drug on the cells.After the intervention of tripterine,the cell protein was extracted,and the expression level of NF-?B and other related proteins in the cells was detected by Western Bloting.Results:(1)After 6 weeks of transfection,the liver tissue of the AKT group had steatosis,and the liver tissue of the mice in the c-Met group was normal,but the liver surface of the AKT/c-Met group was grayish-white with different sizes.Nodular,mouse liver histopathology results showed hepatocellular carcinoma.Compared with the normal group,ALT and AST were significantly increased in the AKT group,c-Met group and AKT/c-Met group,with statistical difference(P<0.05,P<0.01,P<0.001).Compared with the normal group,the expression of NF-?B in the AKT/c-Met group was significantly increased,and the difference was statistically significant(P<0.001).(2)Compared with the model group,the liver weight of the low and high doses of tripterine was decreased(P<0.05).Compared with the model group,the expression of NF-?B protein in liver tissue of low and high doses of tripterine was decreased(P<0.01,P<0.001).(3)The expression level of NF-?B protein in SMMC-7721 cells was lower than that in HepG2 cells.Tripterine inhibited the expression of NF-?B and TNF-? protein in SMMC-7721 and HepG2 cells.Conclusion: Tripterine inhibits the development of the AKT/c-Met-induced HCC mouse model,which may be through inhibition of the NF-?B signaling pathway. |
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