| Objective:1.To establish a golden hamster model of ICC usingBOP[N-nitrosobis(2-oxopropyl)amine].2.To explore the protein expression ofWisp-1、β-catenin、TGF-β1and Smad4in ICC and its relationship with thetumorigenesis.Method:1.30female golden hamsters aged8-9weeks(experimentalgroup27, control group3),experimental group animal subcutaneous injectionof high dose of BOP for5times(weekly, the first dose70mg/kg body weight,and each of the remaining20mg/kg), the control group injected withsaline.17-54days after the first dose of treatment12hamsters of theexperimental group died,4hamsters of the experimental group escaped,ahamster of the control group was killed,the liver was removed and paraffinsections prepared for histopathological observation;54days after the firsttreatment,the remaining surviving(n=13) were killed(n=13,including11of theexperimental group and2of the control group),the liver was removed andparaffin sections and part of the liver was stored in the-80°C refrigerator.2.Gather57cases of Paraffin-embedded blocks(31cases from WuZeshan,who use a high dose of BOP induced hamsters ICC,26cases from thisexperiment use a high dose of BOP induced hamsters ICC),detected theprotein expression of Wisp-1,β-catenin,TGF-β1and Smad4byimmunohistochemistry(IHC) in these blocks.Collected40cases of humanICC Paraffin-embedded blocks,also detected the protein expression of Wisp-1,β-catenin,TGF-β1and Smad4by IHC for comparison.Collected13cases of fresh tissue,detected the protein expression of TGF-β1and Smad4byWestern blotting.Result:1.Most of the animals of the experimental group(14/23)developed ICC,part of the animal(7/23) developed bile duct dysplasia,1developed focal bile duct hyperplasia,1not found bile duct hyperplasia.2.The positive expression rates of Wisp-1、β-catenin and Smad4proteinin both the ICC and the tissue of bile duct dysplasia were higher than normalintrahepatic bile duct tissue(p<0.01), The positive expressions of them in bileduct dysplasia tissue was more likely higher than ICC tissue,but the differencewas no statistical significance(P>0.05).The positive expression rates ofTGF-β1protein in the ICC tissue was higher than normal intrahepatic bileduct tissue(p<0.01),the positive expressions in bile duct dysplasia tissue wasmore likely higher than normal intrahepatic bile duct tissue,but the differencewas no statistical significance(P>0.05)，the positive expressions in the tissueof bile duct dysplasia were lower than ICC tissue(p<0.05).The positiveexpression rates of Wisp-1protein in human ICC tissue was higher than thegolden hamster ICC tissue(P<0.05).The positive expression rates ofβ-cateninand TGF-β1protein in human ICC tissue were more likely higher than thegolden hamster ICC tissue,but the difference was no statisticalsignificance(P>0.05).The positive expression rates of Smad4protein inhuman ICC tissue was more likely lower than the golden hamster ICCtissue,but the difference was no statistical significance(P>0.05).The result ofwestern blotting showed that the protein expression of TGF-β1in ICC tissuewas lower than bile duct dysplasia and normal intrahepatic bile ducttissue(P<0.05),the protein expression in bile duct dysplasia was more likely lower than normal intrahepatic bile duct tissue, but the difference was nostatistical significance(P>0.05).The protein expression of Smad4in ICCtissue was higher than bile duct dysplasia and normal intrahepatic bile ducttissue(P<0.05),the protein expression in bile duct dysplasia was more likelyhigher than normal intrahepatic bile duct tissue, but the difference was nostatistical significance(P>0.05).Conclusion：1.A high dose of BOP can induce a golden hamster modelof ICC,which provides a reliable animal model for the study of ICC,withadvantages of short cycle and high reproducibility.2.Immunohistochemistry showed the protein high expression of Wisp-1and β-catenin in both human intrahepatic cholangiocarcinoma andBOP-induced the golden hamster intrahepatic cholangiocarcinoma, especiallyin the bile duct dysplasia stage,the expression even more likely higher thancholangiocarcinoma,which suggests that Wisp-1and β-catenin play animportant role in the initial stage,and may be closely related to the molecularpathological basis of of ICC.3. Immunohistochemistry showed the protein expression of TGF-β1ingolden hamster ICC was higher than normal intrahepatic bile duct tissue,andit also highly expressed in human ICC tissue,but it lowly expressed in the bileduct dysplasia tissue,the result of western blotting showed that the proteinexpression of TGF-β1in ICC was lower than bile duct dysplasia and normalintrahepatic bile duct tissue, suggesting that the function of TGF-β1in ICCneed further research.4. Immunohistochemistry showed the protein expression of Smad4ingolden hamster ICC and bile duct dysplasia tissue were higher than normalintrahepatic bile duct tissue,and it also highly expressed in human ICC tissue,which suggests that Smad4may be related to the occurrence anddevelopment of ICC.the result of western blotting also showed that theprotein expression of Smad4in ICC and bile duct dysplasia tissue was higherthan normal intrahepatic bile duct tissue,suggesting that Smad4plays animportant role in the development and progression of ICC. |
PDF Full Text Request