AIM:To investigate the role of caveolin-3(CAV3)and cholecystokinin A receptor(CCKAR)in cholesterol gallstone disease(CGD).METHODS:To establish a mouse model of CGD,male C57BL/6 mice were fed with a lithogenic diet con...AIM:To investigate the role of caveolin-3(CAV3)and cholecystokinin A receptor(CCKAR)in cholesterol gallstone disease(CGD).METHODS:To establish a mouse model of CGD,male C57BL/6 mice were fed with a lithogenic diet containing 1.0%cholic acid,1.25%cholesterol and 15%fat;a similar control group was given a normal diet.The fresh liver weights and liver-to-body weight ratio were compared between the two groups after one month.Serum lipid profile and bile composition were determined with an autoanalyzer.The Cav3 and Cckar mRNA and CAV3and CCKAR protein levels in the liver and gallbladder were determined via real-time polymerase chain reaction and Western blot,respectively.RESULTS:Establishment of the mouse CGD model was verified by the presence of cholesterol gallstones in mice fed the lithogenic diet.Compared with mice maintained on a normal diet,those fed the lithogenic diet had significantly higher mean liver-to-body weight ratio(0.067±0.007 vs 0.039±0.007,P<0.01),serum total cholesterol(4.22±0.46 mmol/L vs 2.21±0.11 mmol/L,P<0.001),bile total cholesterol(1.33±0.33 mmol/L vs 0.21±0.11 mmol/L,P<0.001),and bile phospholipid concentrations(3.55±1.40 mmol/L vs 1.55±0.63 mmol/L,P=0.04),but lower total bile acid concentrations(726.48±51.83μmol/L vs 839.83±23.74μmol/L,P=0.007).The lithogenic diet was also associated with significantly lower CAV3 in the liver and lower CAV3 and CCKAR in the gallbladder compared with the control mice(all P<0.05).CONCLUSION:CAV3 and CCKAR may be involved in cholesterol gallstone disease.展开更多
Objective:To discuss the abnormal expression of Wnt inhibitory factor(WIFI) in hepatocellular carcinoma cells and its regulating effect on the hepatocellular carcinoma invasion and metastasis factors of tissue inhibit...Objective:To discuss the abnormal expression of Wnt inhibitory factor(WIFI) in hepatocellular carcinoma cells and its regulating effect on the hepatocellular carcinoma invasion and metastasis factors of tissue inhibitor of matrix metalloproteinases-3(TIMP-3)and caveolin-1.Methods:RT-PCR and Western blot were employed to detect the expression of WIF1 in six hepatocellular carcinoma eell lines of HepG2,Hep3 B,Huh7,PLC/PRF/5.SMMC-7721 and MHCC97 and the immortalized human liver cell line THLE-3.Besides,Lipofectamine 2000 was employed to transfect the eukaryotic expression vector pcDNA3.lWIF1 and blank plasmid pcDNA3.1 into hepatocellular carcinoma cell lines.Transwell assay was used to detect the effect of WIF1 on the invasion ability of hepatocellular carcinoma cells;Western blot was used to detect the effect of WIF1 on the expression of TIMP-3 and caveolin-1in hepatocellular carcinoma cells,it also discussed the effect on the expression of β-catenin.Results:The expression of WIF1 in hepatocellular carcinoma cell lines was lower than that in the normal liver cell lines(P<0.01);while there was basically no expression of WIF1 in the human highly metastatic cell line MHCC-97 and moderate expression in HepG2 and SMMC-7721.Therefore,HepG2 and SMMC-7721 were chosen as the further experimental cell lines.After transfecting the eukaryotic expression vector peDNA3.1-WEFl and blank plasmid pcDNA3.1 into hepatocellular carcinoma eell lines,compared with(he blank plasmid group,the cell viability and invasion ability in the WIF1 group were all reduced(P<0.01),the expression of TIMP-3,caveolin-1 and mRNA were all down-regulated(P<0.01),and the expression of β-catenin was decreased(P<0.01).Conclusions:Because of down-regulation or missing of expression of WIFI in hepatocellular carcinoma cell lines,the up-regulation of WIFI expression can significantly inhibit the invasion and metastasis of HepG2 and SMMC-7721 of hepatocellular carcinoma cell lines,which are related to the up-regulated expression of TIMP-3 and down-regulated expression of caveolin-1 and may be realized through the Wnt/β-catenin signaling pathway.展开更多
基金Supported by National Natural Science Foundation of China,No.81070366
文摘AIM:To investigate the role of caveolin-3(CAV3)and cholecystokinin A receptor(CCKAR)in cholesterol gallstone disease(CGD).METHODS:To establish a mouse model of CGD,male C57BL/6 mice were fed with a lithogenic diet containing 1.0%cholic acid,1.25%cholesterol and 15%fat;a similar control group was given a normal diet.The fresh liver weights and liver-to-body weight ratio were compared between the two groups after one month.Serum lipid profile and bile composition were determined with an autoanalyzer.The Cav3 and Cckar mRNA and CAV3and CCKAR protein levels in the liver and gallbladder were determined via real-time polymerase chain reaction and Western blot,respectively.RESULTS:Establishment of the mouse CGD model was verified by the presence of cholesterol gallstones in mice fed the lithogenic diet.Compared with mice maintained on a normal diet,those fed the lithogenic diet had significantly higher mean liver-to-body weight ratio(0.067±0.007 vs 0.039±0.007,P<0.01),serum total cholesterol(4.22±0.46 mmol/L vs 2.21±0.11 mmol/L,P<0.001),bile total cholesterol(1.33±0.33 mmol/L vs 0.21±0.11 mmol/L,P<0.001),and bile phospholipid concentrations(3.55±1.40 mmol/L vs 1.55±0.63 mmol/L,P=0.04),but lower total bile acid concentrations(726.48±51.83μmol/L vs 839.83±23.74μmol/L,P=0.007).The lithogenic diet was also associated with significantly lower CAV3 in the liver and lower CAV3 and CCKAR in the gallbladder compared with the control mice(all P<0.05).CONCLUSION:CAV3 and CCKAR may be involved in cholesterol gallstone disease.
基金supported by National Science&Technology Pillar Program during the 12th Five-year Plan Period under the research and development of new trisacryl gelatin microspheres(No.2012BAH5B06)
文摘Objective:To discuss the abnormal expression of Wnt inhibitory factor(WIFI) in hepatocellular carcinoma cells and its regulating effect on the hepatocellular carcinoma invasion and metastasis factors of tissue inhibitor of matrix metalloproteinases-3(TIMP-3)and caveolin-1.Methods:RT-PCR and Western blot were employed to detect the expression of WIF1 in six hepatocellular carcinoma eell lines of HepG2,Hep3 B,Huh7,PLC/PRF/5.SMMC-7721 and MHCC97 and the immortalized human liver cell line THLE-3.Besides,Lipofectamine 2000 was employed to transfect the eukaryotic expression vector pcDNA3.lWIF1 and blank plasmid pcDNA3.1 into hepatocellular carcinoma cell lines.Transwell assay was used to detect the effect of WIF1 on the invasion ability of hepatocellular carcinoma cells;Western blot was used to detect the effect of WIF1 on the expression of TIMP-3 and caveolin-1in hepatocellular carcinoma cells,it also discussed the effect on the expression of β-catenin.Results:The expression of WIF1 in hepatocellular carcinoma cell lines was lower than that in the normal liver cell lines(P<0.01);while there was basically no expression of WIF1 in the human highly metastatic cell line MHCC-97 and moderate expression in HepG2 and SMMC-7721.Therefore,HepG2 and SMMC-7721 were chosen as the further experimental cell lines.After transfecting the eukaryotic expression vector peDNA3.1-WEFl and blank plasmid pcDNA3.1 into hepatocellular carcinoma eell lines,compared with(he blank plasmid group,the cell viability and invasion ability in the WIF1 group were all reduced(P<0.01),the expression of TIMP-3,caveolin-1 and mRNA were all down-regulated(P<0.01),and the expression of β-catenin was decreased(P<0.01).Conclusions:Because of down-regulation or missing of expression of WIFI in hepatocellular carcinoma cell lines,the up-regulation of WIFI expression can significantly inhibit the invasion and metastasis of HepG2 and SMMC-7721 of hepatocellular carcinoma cell lines,which are related to the up-regulated expression of TIMP-3 and down-regulated expression of caveolin-1 and may be realized through the Wnt/β-catenin signaling pathway.
