Effect And Mechanism Of Matrine Combined With 5-FU On Liver Cancer

Background:Hepatocellular carcinoma(HCC)is one of the malignancies with high morbidity and mortality rates worldwide,imposing a significant burden on patients and society.Traditional treatment modalities,such as surgical resection,radiotherapy,and chemotherapy,have limited effectiveness and safety in patients with advanced HCC.In recent years,the development of novel anti-tumor drugs and combination chemotherapy regimens has made some progress,providing new therapeutic insights for improving the survival rate and quality of life for HCC patients.5-Fluorouracil(5-FU)is a widely used anti-cancer drug in clinical oncology that inhibits tumor cell growth by interfering with DNA and RNA synthesis.However,the efficacy of 5-FU monotherapy in HCC treatment is limited,and it may cause significant side effects.Therefore,exploring new combination treatment strategies has become a research focus.Matrine is an alkaloid compound derived from the traditional Chinese herbal medicine Sophora flavescens.Studies have found that matrine possesses various biological activities,such as anti-inflammatory,antioxidant,and anti-tumor properties,and it exerts inhibitory effects on a variety of malignant tumor cells.Based on the individual anti-tumor effects and mechanisms of action of matrine and 5-FU,this study proposes a combination therapy for HCC using both agents.Theoretically,the combined use of matrine and 5-FU may exert synergistic effects,enhancing therapeutic outcomes while mitigating the side effects of single-agent treatment.However,there is limited clinical research on the combination of matrine and 5-FU for HCC treatment,and data regarding its efficacy and safety are urgently needed.Objective: This study aims to investigate the effects and molecular mechanisms of matrine combined with 5-FU in the treatment of hepatocellular carcinoma.Methods:MTT assay was used to detect the effect of matrine combined with 5-FU on the proliferation of hepatocellular carcinoma(HCC)Hep G2 cells,measuring the OD value and calculating the combination index(CI)value.A plate colony formation assay was employed to evaluate the synergistic inhibitory effect of matrine combined with 5-FU on HCC cell migration.A cell scratch assay was used to examine the effect of combination treatment on cell migration.Flow cytometry was employed to assess the effect of combination treatment on the cell cycle of HCC cells.A transwell assay was conducted to evaluate the influence of combination treatment on HCC cell migration and invasion.Western blot analysis was utilized to investigate the effect of drug treatment on the expression of apoptosis,cell cycle,and migration-related proteins in HCC cells.Western blot analysis was employed to evaluate the effect of combination treatment on the MAPK p38 signaling pathway.Fluorescence analysis was conducted to examine the effect of combination treatment on JC-1 in HCC cells.The p38 MAPK signaling pathway agonist was introduced,and the impact of combination treatment on HCC cell proliferation,migration,invasion,and cell cycle after pathway activation was evalua.A subcutaneous xenograft model in nude mice was used to evaluate the inhibitory effect of matrine combined with 5-FU on Hep G2 cells in vivo.Results:1.The CI values of matrine combined with 5-FU in the treatment of hepatocellular carcinoma cells were all less than 1,and the differences were statistically significant.2.Plate colony formation assay demonstrated that matrine combined with 5-FU significantly inhibited the colony formation rate of HCC cells,and the difference was statistically significant.3.Flow cytometry results and Western blot analysis of cell cycle protein levels showed that matrine combined with 5-FU arrested Hep G2 cells at the S phase,and the difference was statistically significant.4.Western blot analysis revealed that combination treatment upregulated cleaved-PARP,cleaved-caspase 3,and cleaved-caspase 7 expression,and the difference was statistically significant.5.Cell scratch assay showed that matrine combined with 5-FU reduced the 48 h cell migration rate of Hep G2 cells,and the difference was statistically significant.6.Transwell assay results demonstrated that matrine combined with 5-FU decreased the number of Hep G2 cells passing through the membrane at 48 h,and the difference was statistically significant.7.Western blot analysis results revealed that combination treatment downregulated the expression levels of cell growth factors tgf-β and cleaved-tgf-β,and the difference was statistically significant.8.Western blot analysis results showed that matrine combined with 5-FU could affect the phosphorylation of the p38MAPK/Akt signaling pathway.9.After introducing a specific p38 MAPK signaling pathway agonist,MTT,plate colony formation assay,cell cycle,cell scratch assay,and Western blot analysis results showed that the agonist reduced the anti-proliferative,anti-migratory,and pro-apoptotic effects of matrine combined with 5-FU on Hep G2 cells.10.JC-1 staining experiment results revealed that matrine combined with 5-FU induced mitochondrial damage in Hep G2 cells.11.Subcutaneous xenograft experiment results in nude mice demonstrated that matrine combined with 5-FU significantly inhibited tumor growth in vivo without causing additional liver damage.Conclusions:1.Matrine combined with 5-FU exhibited dose-dependent inhibition of Hep G2 cell proliferation.2.Matrine combined with 5-FU exerts a synergistic anti-hepatocellular carcinoma effect both in vitro and in vivo by inhibiting Hep G2 cell proliferation,migration,and inducing apoptosis.3.The primary mechanism of action for the anti-hepatocellular carcinoma effect of matrine combined with 5-FU is the regulation of p38MAPK/Akt signaling pathway expression and the induction of mitochondrial damage within cells.