ABSTRACT
Objective To establish a luciferase labeled McA-RH7777 hepatoma rat model,which could be used for gross observation to further observe the effect of selective ligation of the portal vein and bile duct on tumor growth and metastasis.Methods The luciferase gene was transfected into rat McA-RH7777 hepatoma cells with pCDH-puromycin-CMV as the carrier,which were subcutaneously inoculated into Buffalo rats.Tumor pieces were then heterotransplanted into the left lateral lobe of the allogenic rat liver to observe the tumor growth in vivo.After the successful hepatoma modeling,the rats were randomly divided into three groups,namely the implanted portal vein group with combined portal vein and bile duct ligation,the implanted portal vein group with single portal vein ligation and sham operation group.The rats were executed at the 1 st week and 2nd week after ligation,and the livers were dissected to record the tumor growth and metastasis inside and outside the liver,respectively.Results The tumor formation rates of Buffalo rats after subcutaneous and intrahepatic implantation were both 100％.The fluorescence signal implanted into the liver lobe could be observed in vivo after the intrahepatic implantation of luciferase transfected Luc-McA-RH7777 at 2nd week,the range and intensity of which increased over time.Only local tumor growth could be found at the 4th week,without obvious intrahepatic and lung metastasis.However,both an increased in situ tumor volume and the pulmonary metastasis could be observed in the implanted portal vein group with combined portal vein and bile duct ligation at 2nd week after the ligation.Immunohistochemistry showed AFP positive immunoreactions in the vast majority of intrahepatic tumor cells and Luc positive immunoreactions in part of tumor cells.Conclusion Luc-McA-RH7777 cells could be used to establish the heptoma rat model and the in vivo analysis within the Buffalo rat liver demonstrated that the combined ligation of the portal vein and bile duct can accelerate the development and metastasis of liver cancer.
ABSTRACT
PURPOSE: The proliferative activity of cells in enzyme altered fdegrees Ci of the rat hepatoma model was measured by double immunohistdegrees Chemical staining methods using anti-bromodeoxyuridine (BrdU) and anti-glutathione S transferase of placental form (GST-P). The aim of this study was to compare the cell proliferative activity in GST-P positive altered fdegrees Ci and in negative fdegrees Ci. MATERIALS AND METHODS: Eight-week-old male Sprague-Dawley rats were administered by 200 mg/kg diethylnitrosamine (DEN) intraperitoneally, and followed by 0.02% acetylaminofluorene (AAF)-containing diet for 4 weeks. One week after administration of AAF diet, two-thirds hepa tectomy was performed. Control animals were treated as same except for the omission of AAF in the diet. The rats were sacrified 0, 1, 3, 5, 7, 14 and 21 days after partial hepatectomy. The slices of liver were fixed in acetone, dehydrated in benzene and stained by peroxidase-anti peroxidase method against GST-P and by avidine-biotin peroxidase complex method against BrdU. RESULTS: The area of the GST-P positive fdegrees Ci was increased during the experimental period. In the experimental group, the S-phase fraction in the fdegrees Ci remained high during the first week and was decreased thereafter. However, the GST-P negative area maintained a low S-phase cell frac tion throughout the experimental period. CONCLUSION: These results suggest that hepatic cells in the enzyme altered fdegrees Ci may escape a suppressor effect of AAF in contrast to the normal cells in which their growth are inhibited by AAF.