The Inhibitory Effect Of ERF3b-36 Fragment On Chemically Induced Hepatocarcinoma And Its Mechanism

Objective:To study the inhibitory effect of eRF3b-36 fragment on chemically induced hepatocarcinoma in mice and preliminarily explore its mechanism.Methods:40 male C57BL/6J mice were randomly divided into the normal group,model group,eRF3b-36 fragment 150 and 250?g/kg groups.Each group contained 10 mice.All groups except the normal group were induced hepatocarcinoma with DEN+CCL₄+ethanol for 20 weeks.One day before giving the chemical carcinogen,the eRF3b-36 fragment 150 and250?g/kg groups were injected with the corresponding doses of the eRF3b-36fragment via tail vein twice per week lasting for 20 weeks.At the end of 20th week,all mice were sacrificed.The serum was collected for liver function tests.The livers were harvested and the development of liver tumors in mice were recorded.The pathological changes of liver tissues were detected by HE staining and Sirius red staining.The relative mRNA expression levels of hepatocarcinoma biomarkers such as AFP,GPC3 and cell proliferation related factors such as Ki-67,Myc,Cyclin D1,Cyclin E1 and inflammatory related factors such as TNF-?,TGF-?1 in liver tissues were detected by qRT-PCR.Measurement data were expressed as mean±SD.One-way ANOVA or Kruskal Wallis test were used for multiple comparisons.Statistical significance was taken at the P<0.05 level.Results:1.Liver function indexes:The serum ALT and AST levels in the normal group?40.90±11.64,126.90±25.64?,eRF3b-36 fragment 250?g/kg group?3360.00±1593.88,2333.75±1057.61?were all significantly lower than those in the model group?5157.50±1531.93,3751.25±886.40??P<0.05?.2.The development of liver tumors:Liver tumors were observed in the other three groups except the normal group.And the number of liver tumors in the eRF3b-36 fragment 150?g/kg group?3.38±1.77?and 250?g/kg group?2.88±1.96?were all significantly lower than that in the model group?5.38±2.00??P<0.05?.3.Pathological changes of liver tissue:HE staining showed that there was no pathological change of liver tissues in the normal group.HCC lesions and the obvious cellular atypia were observed in the liver tissues of the model group.The degree of cellular atypia in the eRF3b-36 fragment 150 and250?g/kg groups were lighter than that in the model group.According to the Sirius red staining,there was no fibrosis in the liver tissues of the normal group.And there was obvious fibrosis in the liver tissues of the model group.The degree of liver fibrosis in the eRF3b-36 fragment 150 and 250?g/kg groups were significantly lighter than that in the model group.4.The relative mRNA expression levels of hepatocarcinoma biomarkers:The relative mRNA expression levels of AFP in the normal group?1.00±0.12?and eRF3b-36 fragment 250?g/kg group?8.63±3.11?were all significantly lower than that in the model group?97.33±89.91??P<0.01?.The relative mRNA expression levels of GPC3 in the normal group?1.00±0.40?,eRF3b-36fragment 150?g/kg group?22.10±6.33?and 250?g/kg group?17.77±9.87?were all significantly lower than that in the model group?297.11±256.79??P<0.01?.5.The relative mRNA expression levels of cell proliferation related factors:The relative mRNA expression levels of Ki-67,Myc,Cyclin D1 in the normal group?1.00±0.32,1.00±0.44,1.00±0.22?and eRF3b-36 fragment250?g/kg group?3.23±0.62,5.44±1.15,6.95±3.05?were all significantly lower than those in the model group?5.01±1.23,15.27±4.10,11.30±1.03??P<0.05?.The relative mRNA expression levels of Cyclin E1 in the normal group?1.00±0.40?,eRF3b-36 fragment 150?g/kg group?8.51±2.57?and250?g/kg group?3.47±1.55?were all significantly lower than that in the model group?15.00±5.20??P<0.05?.6.The relative mRNA expression levels of inflammatory related factors:The relative mRNA expression levels of TNF-?in the normal group?1.00±0.27?and eRF3b-36 fragment 250?g/kg group?3.45±1.05?were all significantly lower than that in the model group?5.87±0.99??P<0.01?.The relative mRNA expression levels of TGF-?1 in the normal group?1.00±0.16?,eRF3b-36 fragment 150?g/kg group?1.72±0.34?and 250?g/kg group?1.63±0.52?were all significantly lower than that in the model group?2.68±0.74??P<0.05?.Conclusions:1.The inhibitory effect of eRF3b-36 fragment on chemically induced hepatocarcinoma in mice is mainly manifested in protecting liver function,reducing the number of liver tumors,improving the pathological changes in liver tissues,and reducing the expression of hepatocarcinoma biomarkers AFP and GPC3.2.The mechanism of eRF3b-36 fragment inhibiting chemically induced hepatocarcinoma in mice may be related to down-regulating the expression of cell proliferation and inflammation related factors Ki-67,Myc,Cyclin D1,Cyclin E1,TNF-?and TGF-?1.