OBJECTIVE To explore the role of gecko crude peptides(GCPs)in the proliferation,apoptosis,migration and lymphangiogenesis of human hepatocellular carcinoma cells(Hep G2)and human lymphaticendothelial cells(HLECs)in vi...OBJECTIVE To explore the role of gecko crude peptides(GCPs)in the proliferation,apoptosis,migration and lymphangiogenesis of human hepatocellular carcinoma cells(Hep G2)and human lymphaticendothelial cells(HLECs)in vitro.METHODS The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay was used to evaluate the anti-proliferative effect of GCPs and si RNA-VEGF-C on Hep G2 cells,Hoechst 33258 staining and flow cytometry were performed to analyze cycle and apoptosis.The migration and invasion ability of cells were assayed by transwell chamber experiment and wound-healing assay.The protein and m RNA expressions of vascular endothelial growth factor-C(VEGF-C)and CXC chemokine receptor-4(CXCR4)were detected by q-PCR,immunofluorescence,Western blot.The protein expressions of the extracellular signal regulated kinase(ERKI/2),c-Jun N-terminal kinase(JNK),p38-mitogen activated protein kinases(p38 MAPK),serine/threonine kinase(Akt)and phosphatidylinositol-3-kinase(PI3K)were detected by western blot.The anti-lymphangiogenesis effect of GCPs on the HLECs was analyzed using an in vitro tube-formation assay.The protein and m RNA expressions of vascular endothelial growth factor receptor-3(VEGFR-3)and stromal cell-derived factor-1(SDF-1)were detected by q-PCR,Western blot.RESULTS GCPs and si RNA-VEGF-C inhibited Hep G2 proliferation,invasion and migration,and the most obvious inhibitory effect was both synergistic effects.Thus,GCPs suppressed HLECs proliferation,migration and tubelike structure formationin a dose-dependent manner,and had inhibitory effect of tumor-induced lymphangiogenesis in vitro.Additionally,we found that GCPs and si RNA-VEGF-C decreased the expressions of MMP-2,MMP-9,VEGF-C,CXCR4,phospho-ERK1/2,phospho-P38,phospho-JNK and PI3K in Hep G2 cells.Moreover,GCPs had a dose-dependent depressive effecton the expressions of VEGFR-3,SDF-1 in HLECs.CONCLUSION The low expression of VEGF-C mediated by si RNA-VEGF-C and GCPs inhibit tumor proliferation,invasion and migrationby suppressing the MAPK signaling pathway through reduced levels of VEGF-C,and GCPs inhibit tumor lymphangiogenesis by suppressing the CXCR4/SDF-1 signaling pathway through suppressed VEGF-C/VEGFR-3.展开更多
OBJECTIVE Angiogenesis therapy has attracted interest as a potential treatment for hepatocellular carcinoma(HCC).In this study,we investigated the anti-proliferative activities and antiangiogenesis effects of saikosap...OBJECTIVE Angiogenesis therapy has attracted interest as a potential treatment for hepatocellular carcinoma(HCC).In this study,we investigated the anti-proliferative activities and antiangiogenesis effects of saikosaponins(SS)-b on hepatocellular carcinoma(HCC)and its regulation on VEGF/ERK/HIF-1 αsignal pathway.METHODS H22 hepatoma-bearing mice model and HepG-2 cells were used to study the anti-tumor and anti-angiogenesis effects of SS-b in vivo and in vitro.Pathological change of tumor tissue was observed by HE staining,the microvascular changes were detected by immunohistochemical method.The effects of SS-b on angiogenesis were examined by using the chick embryo chorioallantoic membrane(CAM)model.The effects of SS-b on proliferation,migration and invasion were investigated by MTT assay,scratch wound healing assay and transwell assay inhuman umbilical vein endothelial cell(HUVEC)and HepG2 cells in vitro.Vascular endothelial growth factor(VEGF),matrix metalloproteinase-2/9(MMP-2/9),hypoxia-inducible factor-1α(HIF-1α)expression and the phosphorylation of extracellular regulated kinase(ERK)were analyzed using RT-PCR and Westernblot.RESULTS SS-b effectively inhibited the tumor growth of H22 mice in vivo.The inhibitory rate of tumor was 49.1%,50.7%,66.1%in SS-b 5,10 and 20 mg·kg-1group respectively.HE staining results showed that SS-b induced tumor necrosis and nuclear dissolution in H22 mice.Moreover,SS-b also reduced the number of microvessels of tumor tissue in H22 mice significantly and suppressed the angiogenesis of CAM induced by b-FGF.SS-b had an obvious inhibitory effect on cell proliferation,migration and invasion of HUVEC cells and HepG-2 cells.These effects were associated with downregulation of the expression of MMP2/9 and suppression of VEGF/ERK/HIF-1αsignaling in H22 mice and Hep-G2 cells.CONCLUSION Our findings showed that SS-b exerts anti-tumor effects by inhibiting tumor angiogenesis via regulating VEGF/ERK/HIF-1α signal pathway in vivo and in vitro.展开更多
OBJECTIVE Metastasis-associated in colon cancer-1(MACC1)is an oncogene that has been newly identified.It promotes tumor proliferation and invasion via the MET pathway.Our study investigated the effects of Saikosaponin...OBJECTIVE Metastasis-associated in colon cancer-1(MACC1)is an oncogene that has been newly identified.It promotes tumor proliferation and invasion via the MET pathway.Our study investigated the effects of Saikosaponin-b(SS-b)on the proliferation and apoptosis of HepG2 cells and its regulation on MACC1/c-Met/Akt signaling pathway.METHODS HepG2 cells were treated with SS-b(10-800 g·L^(-1))for 48 h in vitro.The CCK-8 assay was used to assess cell proliferation,and cell apoptosis was determined by Hoechst33258 staining,AnnexinⅤ/PI staining and caspase 3 assay.RT-PCR was used to examine the expression of MACC1,c-MET and hepatocyte growth factor(HGF)mR NA.MACC1 protein was detected by Western blot and immunohistochemistry.The protein expressions of p-cMET,c-MET,p-AKT,AKT,p-BAD,BAD were measured by Western blot.RESULTS SS-b inhibited the growth of HepG2 cells in dose-dependent way and induced cell apoptosis significantly.HepG2 cells showed karyopyknosis,fragmentation and fluorescence highlight in SS-b treatment group.FCM results showed that apoptosis rate of HepG2 cells increased with SS-b concentration.The immunofluorescence results showed that the MACC1 expression decreased significantly in HepG2 cells treated with SS-b.The expression levels of MACC1,c-MET and HGF mR NA in HepG2 cells were significantly inhibited by SS-b.SS-b also significantly decreased the protein expressions of MACC1,p-c-MET and p-AKT while increased the expression of p-BAD and caspase 3 in HepG2 cells(P<0.05).CONCLUSION SS-b inhibited the proliferation and induced the apoptosis of HepG2 cells by targeting the MACC1/c-Met/Akt signaling pathway.展开更多
OBEJECTIVE Gecko has been clinically used in China for many years.It has been proved that the gecko polypeptide mixture(GPM) extracted from gecko could inhibit the growth of multiple types of tumor cells.In order to i...OBEJECTIVE Gecko has been clinically used in China for many years.It has been proved that the gecko polypeptide mixture(GPM) extracted from gecko could inhibit the growth of multiple types of tumor cells.In order to investigate the possible anti-tumor molecular mechanisms of GPM,we used RNA-seq technology to identify the differentially expressed genes of human hepatocellular carci.noma(HCC) HepG2 cells treated with or without GPM.METHODS The HepG2 cells were treated with different concentration of GPM(0,0.1,0.2,0.3,0.4 mg·mL-1) for 6 h,12 h and 24 h,respectively.MTT assay was used to detect the viability of HepG2 cells.DAPI fluorescence staining was performed to observe nucleus morphological changes of HepG2 cells.Western blot analysis was applied to observe the expres.sion of apoptosis-related proteins and endoplasmic reticulum stress(ERs)-related proteins in HepG2 cells.Flow cytometry was also applied to detect reactive oxygen species(ROS) generation.In this report,we showed that GPM could induce HepG2 cells apoptosis and influence HepG2 cells proliferation in a dose-dependent manner.We applied many analysis methods,including differentially expressed genes analysis,Gene Ontology(GO) enrichment analysis,KEGG pathway enrichment analysis,protein-protein interaction network analysis to screen out possible molecular mechanisms.RESULTS ER-nucleus signaling pathway,cellular response to stress and apoptotic processes were identified the potential anti-cancer molecular biological process of GPM.GPM may also induce apoptosis in HepG2 cells via endoplasmic reticulum stress pathway.The GPM could induce ROS generation and up-regulate ERs-related proteins.CONCLUSION The present study revealed the potential anti-tumor mechanism of GPM.展开更多
OBJECTIVE To explore the therapeutic effects and underlying mechanisms of velvet antler polypeptides(VVAPs)in CCl4-induced experimental hepatic fibrosis in rats.METHODS Anti-hepatic fibrosis properties of VAPs were te...OBJECTIVE To explore the therapeutic effects and underlying mechanisms of velvet antler polypeptides(VVAPs)in CCl4-induced experimental hepatic fibrosis in rats.METHODS Anti-hepatic fibrosis properties of VAPs were tested by Subcutaneous injection(SC)into male Wistar rats of CCl4- induced experimental hepatic fibrosis.After SC injections for 45 consecutive days at doses of 5mg·kg-1(low dose,VAPsL),10mg·kg-1(mid-dose,VAPsM)and 20mg·kg-1(high-dose,VAPsH),the rats were sacrificed and the various indicators were evaluated and tested.Observed hepatic cells degeneration and necrosis,inflammatory infiltration and levels of serum enzymes to assess treatment of VAPs;The expression levels of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),MDA,and hydroxyproline(HYP)in liver tissue were analyzed;RT-PCR analysis was carried out to detect the expression levels of matrix metalloproteinases2(MMP-2)and tissue inhibitor of metalloproteinases 1(TIMP-1)in liver tissue.RESULTS VAPs has obvious anti-hepatic fibrosis effects.Hepatocyte swelling,fatty degeneration was significantly reduced,reducing infiltration of inflammatory cells.Release of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)decreased significantly,reduction of hyaluronic acid(HA)and laminin(LN)obviously,at the same time,the content of total protein and albumin increased significantly in serum.Activity of SOD and GSH-Px was significantly raised and the content of MDA and HYP was reduced significantly in liver tissue.Expression levels of MMP-2and TIMP-1 mRNA in liver were decreased significantly.These improvements were more significant in high-dos and mid-dose groups(P<0.05 or P<0.01 vs model group).CONCLUSION These findings suggest VAPs can significant treat the hepatic fibrosis,which may be due to protect liver cells and improve liver functions by hydroxyl radical scavenging activity and great effect of antioxidation,and decrease the gene expression of MMP-2,improving exist-environment of liver cells and decreasing the gene expression of TIMP-1,prompting degradation of extracellular matrix.展开更多
OBJECTIVE In order to investigate the possible anti-tumor molecular mechanisms of gecko polypeptide mixture(GPM).METHODS RNA-seq technology was used to identify the differentially expressed genes of human hepatocellul...OBJECTIVE In order to investigate the possible anti-tumor molecular mechanisms of gecko polypeptide mixture(GPM).METHODS RNA-seq technology was used to identify the differentially expressed genes of human hepatocellular carcinoma(HCC)HepG2 cells treated with or without GPM.The HepG2 cells were treated with different concentration of GPM(0,0.1,0.2,0.3,0.4 mg·mL^(-1))for 6 h,12 h and 24 h,respectively.MTT assay was used to detect the viability of HepG2 cells.DAPI fluorescence staining was performed to observe nucleus morphological changes of HepG2 cells.Western blot analysis was applied to observe the expression of apoptosis-related proteins in HepG2 cells.RESULTS The results showed that GPM could induce HepG2 cells apoptosis and influence HepG2 cells proliferation in a dose-dependent manner.We applied many analysis methods,including differentially expressed genes analysis,Gene Ontology(GO)enrichment analysis,KEGG pathway enrichment analysis,protein-protein interaction network analysis to screen out possible molecular mechanisms.ER-nucleus signaling pathway,cellular response to stress and apoptotic processes were identified the potential anti-cancer molecular biological process of GPM.GPM may also induce apoptosis in HepG2 cells via endoplasmic reticulum stress pathway.The mechanism is closely related to ERs,which might be beneficial for clinical therapy of HCC.CONCLUSION GPM can inhibit cells proliferation and induce apoptosis in HepG2 cells.The gene expression profile of GPM in HepG2 cells was obtained.The present study revealed the potential anti-tumor mechanism of GPM.展开更多
OBJECTIVE To investigate the regulation of {O^2(2,4-dinitrophenyl)1-[(4-ethoxycarbonyl)piperazin-1-yl]diazen-1-ium-1,2-diolate}(JS-K),anitric oxide donor,on tumor energy metabolism in H22 tumor-bearing mice.METHODS Th...OBJECTIVE To investigate the regulation of {O^2(2,4-dinitrophenyl)1-[(4-ethoxycarbonyl)piperazin-1-yl]diazen-1-ium-1,2-diolate}(JS-K),anitric oxide donor,on tumor energy metabolism in H22 tumor-bearing mice.METHODS The hepatoma animal model in BALB/c mice was established with H22 cell line.The JS-K group and model group were received JS-K(0.75 and 1.5 mg?kg^(-1)) and saline via tail intravenous once every 3 d for 14 d,received 5 injections,respectively.The positive group was received 5-FU 20 mg·kg^(-1) by intraperitoneal injection once a day for 14 d.On the 15 th day mice were sacrificed.The tumor growth inhibition rate were calculated.The activities of hexokinase(HK),phosphofructo kinase(PFK),pyruvate kinase(PK),succinate dehydrogenase(SDH),adenosine triphosphatase(ATPase),and the levels of lactic acid(LD) and adenosine triphosphate(ATP) in tumor tissues were determined by colorimetric method.RESULTS Compared with model group,the tumor mass of JS-K0.75 and 1.5 mg·kg^(-1) group was significantly reduced(P<0.01),and the tumor growth inhibition rate was 23.9% and 50.3%,respectively.The activity of HK,PFK,PK,SDH and ATPase of tumor tissue in model group was(22.6±3.7,14.4±2.6,12.9±3.2 and 10.5±2.6)U·g^(-1) protein and(0.70±0.10)μmol Pi·mg^(-1) protein per hour,respectively;which in JS-K 1.5 mg?kg^(-1) group was dropped by 42.0%,26.6%,22.7%,23.3% and 21.7%(P<0.01,P<0.05).Compared with the model group,the level of ATP and LD in JS-K group was dropped(P<0.01).CONCLUSION JS-K can inhibit the growth of tumor in H22 tumor-bearing mice and its mechanism may be related to regulating the tumor energy metabolism with inhibition of glycolysis and aerobic oxidation.展开更多
基金supported by Medical Science and Technology Research Project of Henan Province(142102310031)
文摘OBJECTIVE To explore the role of gecko crude peptides(GCPs)in the proliferation,apoptosis,migration and lymphangiogenesis of human hepatocellular carcinoma cells(Hep G2)and human lymphaticendothelial cells(HLECs)in vitro.METHODS The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay was used to evaluate the anti-proliferative effect of GCPs and si RNA-VEGF-C on Hep G2 cells,Hoechst 33258 staining and flow cytometry were performed to analyze cycle and apoptosis.The migration and invasion ability of cells were assayed by transwell chamber experiment and wound-healing assay.The protein and m RNA expressions of vascular endothelial growth factor-C(VEGF-C)and CXC chemokine receptor-4(CXCR4)were detected by q-PCR,immunofluorescence,Western blot.The protein expressions of the extracellular signal regulated kinase(ERKI/2),c-Jun N-terminal kinase(JNK),p38-mitogen activated protein kinases(p38 MAPK),serine/threonine kinase(Akt)and phosphatidylinositol-3-kinase(PI3K)were detected by western blot.The anti-lymphangiogenesis effect of GCPs on the HLECs was analyzed using an in vitro tube-formation assay.The protein and m RNA expressions of vascular endothelial growth factor receptor-3(VEGFR-3)and stromal cell-derived factor-1(SDF-1)were detected by q-PCR,Western blot.RESULTS GCPs and si RNA-VEGF-C inhibited Hep G2 proliferation,invasion and migration,and the most obvious inhibitory effect was both synergistic effects.Thus,GCPs suppressed HLECs proliferation,migration and tubelike structure formationin a dose-dependent manner,and had inhibitory effect of tumor-induced lymphangiogenesis in vitro.Additionally,we found that GCPs and si RNA-VEGF-C decreased the expressions of MMP-2,MMP-9,VEGF-C,CXCR4,phospho-ERK1/2,phospho-P38,phospho-JNK and PI3K in Hep G2 cells.Moreover,GCPs had a dose-dependent depressive effecton the expressions of VEGFR-3,SDF-1 in HLECs.CONCLUSION The low expression of VEGF-C mediated by si RNA-VEGF-C and GCPs inhibit tumor proliferation,invasion and migrationby suppressing the MAPK signaling pathway through reduced levels of VEGF-C,and GCPs inhibit tumor lymphangiogenesis by suppressing the CXCR4/SDF-1 signaling pathway through suppressed VEGF-C/VEGFR-3.
基金supported by Scientific and Technology Projects of Henan Province(142102310137)Science and Technology Development Project of Luoyang City(1603001A-3)
文摘OBJECTIVE Angiogenesis therapy has attracted interest as a potential treatment for hepatocellular carcinoma(HCC).In this study,we investigated the anti-proliferative activities and antiangiogenesis effects of saikosaponins(SS)-b on hepatocellular carcinoma(HCC)and its regulation on VEGF/ERK/HIF-1 αsignal pathway.METHODS H22 hepatoma-bearing mice model and HepG-2 cells were used to study the anti-tumor and anti-angiogenesis effects of SS-b in vivo and in vitro.Pathological change of tumor tissue was observed by HE staining,the microvascular changes were detected by immunohistochemical method.The effects of SS-b on angiogenesis were examined by using the chick embryo chorioallantoic membrane(CAM)model.The effects of SS-b on proliferation,migration and invasion were investigated by MTT assay,scratch wound healing assay and transwell assay inhuman umbilical vein endothelial cell(HUVEC)and HepG2 cells in vitro.Vascular endothelial growth factor(VEGF),matrix metalloproteinase-2/9(MMP-2/9),hypoxia-inducible factor-1α(HIF-1α)expression and the phosphorylation of extracellular regulated kinase(ERK)were analyzed using RT-PCR and Westernblot.RESULTS SS-b effectively inhibited the tumor growth of H22 mice in vivo.The inhibitory rate of tumor was 49.1%,50.7%,66.1%in SS-b 5,10 and 20 mg·kg-1group respectively.HE staining results showed that SS-b induced tumor necrosis and nuclear dissolution in H22 mice.Moreover,SS-b also reduced the number of microvessels of tumor tissue in H22 mice significantly and suppressed the angiogenesis of CAM induced by b-FGF.SS-b had an obvious inhibitory effect on cell proliferation,migration and invasion of HUVEC cells and HepG-2 cells.These effects were associated with downregulation of the expression of MMP2/9 and suppression of VEGF/ERK/HIF-1αsignaling in H22 mice and Hep-G2 cells.CONCLUSION Our findings showed that SS-b exerts anti-tumor effects by inhibiting tumor angiogenesis via regulating VEGF/ERK/HIF-1α signal pathway in vivo and in vitro.
基金supported by Scientific and Technology Projects of Henan Province(142102310137)Science and Technology Development Project of Luoyang City(1603001A-3)
文摘OBJECTIVE Metastasis-associated in colon cancer-1(MACC1)is an oncogene that has been newly identified.It promotes tumor proliferation and invasion via the MET pathway.Our study investigated the effects of Saikosaponin-b(SS-b)on the proliferation and apoptosis of HepG2 cells and its regulation on MACC1/c-Met/Akt signaling pathway.METHODS HepG2 cells were treated with SS-b(10-800 g·L^(-1))for 48 h in vitro.The CCK-8 assay was used to assess cell proliferation,and cell apoptosis was determined by Hoechst33258 staining,AnnexinⅤ/PI staining and caspase 3 assay.RT-PCR was used to examine the expression of MACC1,c-MET and hepatocyte growth factor(HGF)mR NA.MACC1 protein was detected by Western blot and immunohistochemistry.The protein expressions of p-cMET,c-MET,p-AKT,AKT,p-BAD,BAD were measured by Western blot.RESULTS SS-b inhibited the growth of HepG2 cells in dose-dependent way and induced cell apoptosis significantly.HepG2 cells showed karyopyknosis,fragmentation and fluorescence highlight in SS-b treatment group.FCM results showed that apoptosis rate of HepG2 cells increased with SS-b concentration.The immunofluorescence results showed that the MACC1 expression decreased significantly in HepG2 cells treated with SS-b.The expression levels of MACC1,c-MET and HGF mR NA in HepG2 cells were significantly inhibited by SS-b.SS-b also significantly decreased the protein expressions of MACC1,p-c-MET and p-AKT while increased the expression of p-BAD and caspase 3 in HepG2 cells(P<0.05).CONCLUSION SS-b inhibited the proliferation and induced the apoptosis of HepG2 cells by targeting the MACC1/c-Met/Akt signaling pathway.
文摘OBEJECTIVE Gecko has been clinically used in China for many years.It has been proved that the gecko polypeptide mixture(GPM) extracted from gecko could inhibit the growth of multiple types of tumor cells.In order to investigate the possible anti-tumor molecular mechanisms of GPM,we used RNA-seq technology to identify the differentially expressed genes of human hepatocellular carci.noma(HCC) HepG2 cells treated with or without GPM.METHODS The HepG2 cells were treated with different concentration of GPM(0,0.1,0.2,0.3,0.4 mg·mL-1) for 6 h,12 h and 24 h,respectively.MTT assay was used to detect the viability of HepG2 cells.DAPI fluorescence staining was performed to observe nucleus morphological changes of HepG2 cells.Western blot analysis was applied to observe the expres.sion of apoptosis-related proteins and endoplasmic reticulum stress(ERs)-related proteins in HepG2 cells.Flow cytometry was also applied to detect reactive oxygen species(ROS) generation.In this report,we showed that GPM could induce HepG2 cells apoptosis and influence HepG2 cells proliferation in a dose-dependent manner.We applied many analysis methods,including differentially expressed genes analysis,Gene Ontology(GO) enrichment analysis,KEGG pathway enrichment analysis,protein-protein interaction network analysis to screen out possible molecular mechanisms.RESULTS ER-nucleus signaling pathway,cellular response to stress and apoptotic processes were identified the potential anti-cancer molecular biological process of GPM.GPM may also induce apoptosis in HepG2 cells via endoplasmic reticulum stress pathway.The GPM could induce ROS generation and up-regulate ERs-related proteins.CONCLUSION The present study revealed the potential anti-tumor mechanism of GPM.
基金The project supported by National Natural Science Foundation of China(U1204826)
文摘OBJECTIVE To explore the therapeutic effects and underlying mechanisms of velvet antler polypeptides(VVAPs)in CCl4-induced experimental hepatic fibrosis in rats.METHODS Anti-hepatic fibrosis properties of VAPs were tested by Subcutaneous injection(SC)into male Wistar rats of CCl4- induced experimental hepatic fibrosis.After SC injections for 45 consecutive days at doses of 5mg·kg-1(low dose,VAPsL),10mg·kg-1(mid-dose,VAPsM)and 20mg·kg-1(high-dose,VAPsH),the rats were sacrificed and the various indicators were evaluated and tested.Observed hepatic cells degeneration and necrosis,inflammatory infiltration and levels of serum enzymes to assess treatment of VAPs;The expression levels of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),MDA,and hydroxyproline(HYP)in liver tissue were analyzed;RT-PCR analysis was carried out to detect the expression levels of matrix metalloproteinases2(MMP-2)and tissue inhibitor of metalloproteinases 1(TIMP-1)in liver tissue.RESULTS VAPs has obvious anti-hepatic fibrosis effects.Hepatocyte swelling,fatty degeneration was significantly reduced,reducing infiltration of inflammatory cells.Release of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)decreased significantly,reduction of hyaluronic acid(HA)and laminin(LN)obviously,at the same time,the content of total protein and albumin increased significantly in serum.Activity of SOD and GSH-Px was significantly raised and the content of MDA and HYP was reduced significantly in liver tissue.Expression levels of MMP-2and TIMP-1 mRNA in liver were decreased significantly.These improvements were more significant in high-dos and mid-dose groups(P<0.05 or P<0.01 vs model group).CONCLUSION These findings suggest VAPs can significant treat the hepatic fibrosis,which may be due to protect liver cells and improve liver functions by hydroxyl radical scavenging activity and great effect of antioxidation,and decrease the gene expression of MMP-2,improving exist-environment of liver cells and decreasing the gene expression of TIMP-1,prompting degradation of extracellular matrix.
基金supported by Science and Technology Key Research Fund of Henan Province(142102310031)
文摘OBJECTIVE In order to investigate the possible anti-tumor molecular mechanisms of gecko polypeptide mixture(GPM).METHODS RNA-seq technology was used to identify the differentially expressed genes of human hepatocellular carcinoma(HCC)HepG2 cells treated with or without GPM.The HepG2 cells were treated with different concentration of GPM(0,0.1,0.2,0.3,0.4 mg·mL^(-1))for 6 h,12 h and 24 h,respectively.MTT assay was used to detect the viability of HepG2 cells.DAPI fluorescence staining was performed to observe nucleus morphological changes of HepG2 cells.Western blot analysis was applied to observe the expression of apoptosis-related proteins in HepG2 cells.RESULTS The results showed that GPM could induce HepG2 cells apoptosis and influence HepG2 cells proliferation in a dose-dependent manner.We applied many analysis methods,including differentially expressed genes analysis,Gene Ontology(GO)enrichment analysis,KEGG pathway enrichment analysis,protein-protein interaction network analysis to screen out possible molecular mechanisms.ER-nucleus signaling pathway,cellular response to stress and apoptotic processes were identified the potential anti-cancer molecular biological process of GPM.GPM may also induce apoptosis in HepG2 cells via endoplasmic reticulum stress pathway.The mechanism is closely related to ERs,which might be beneficial for clinical therapy of HCC.CONCLUSION GPM can inhibit cells proliferation and induce apoptosis in HepG2 cells.The gene expression profile of GPM in HepG2 cells was obtained.The present study revealed the potential anti-tumor mechanism of GPM.
基金supported by National Natural Science Foundation of China(81502627)the Young Backbone Teachers Assistance Scheme of Henan Province Colleges and Universities(2016GGJS-065)
文摘OBJECTIVE To investigate the regulation of {O^2(2,4-dinitrophenyl)1-[(4-ethoxycarbonyl)piperazin-1-yl]diazen-1-ium-1,2-diolate}(JS-K),anitric oxide donor,on tumor energy metabolism in H22 tumor-bearing mice.METHODS The hepatoma animal model in BALB/c mice was established with H22 cell line.The JS-K group and model group were received JS-K(0.75 and 1.5 mg?kg^(-1)) and saline via tail intravenous once every 3 d for 14 d,received 5 injections,respectively.The positive group was received 5-FU 20 mg·kg^(-1) by intraperitoneal injection once a day for 14 d.On the 15 th day mice were sacrificed.The tumor growth inhibition rate were calculated.The activities of hexokinase(HK),phosphofructo kinase(PFK),pyruvate kinase(PK),succinate dehydrogenase(SDH),adenosine triphosphatase(ATPase),and the levels of lactic acid(LD) and adenosine triphosphate(ATP) in tumor tissues were determined by colorimetric method.RESULTS Compared with model group,the tumor mass of JS-K0.75 and 1.5 mg·kg^(-1) group was significantly reduced(P<0.01),and the tumor growth inhibition rate was 23.9% and 50.3%,respectively.The activity of HK,PFK,PK,SDH and ATPase of tumor tissue in model group was(22.6±3.7,14.4±2.6,12.9±3.2 and 10.5±2.6)U·g^(-1) protein and(0.70±0.10)μmol Pi·mg^(-1) protein per hour,respectively;which in JS-K 1.5 mg?kg^(-1) group was dropped by 42.0%,26.6%,22.7%,23.3% and 21.7%(P<0.01,P<0.05).Compared with the model group,the level of ATP and LD in JS-K group was dropped(P<0.01).CONCLUSION JS-K can inhibit the growth of tumor in H22 tumor-bearing mice and its mechanism may be related to regulating the tumor energy metabolism with inhibition of glycolysis and aerobic oxidation.
