| Objective To establish a hepatoma model in Buffalo rats for monitoring in vivo by using bioluminescent imaging system. We aimed to figure out the role of ligation of bile duct with portal vein, compared to portal vein ligation, in hypertrophy of liver and growth and metastasis of liver cancer, based on such model.Method Luc-McA-RH7777 hepatoma cells were injected subcutaneously into Buffalo rats. Tumor pieces were then heterotransplanted into the left lateral lobe of the allogenic liver. Growth and metastasis of tumor were observed in vivo by using bioluminescent imaging system and ICG surgical navigation system. After successful hepatoma modeling, Buffalo rats were randomly divided into three group, namely the bile duct with portal vein ligation of median lobe and left lateral lobe group(BPL),signal portal vein ligation group(PVL)and sham group. Buffalo rats were executed at 1,3,5,7,14 days after the ligation operation. Liver and lung were dissected for the observation of growth and metastasis of tumors. Then the rats, the liver tissues and tumors were weighed and harvested. The inferior vena caval blood were harvested to measure aminotransferase and bile acid levels. For the non-ligation lobes, Western Blot was used to measure the level of the expression of PCNA and calculate ki-67 positive cells based on immunohistochemistry.Result (1)The tumor incidence of Buffalo rats after subcutaneous and intrahepatic implantation were both 100%.The fluorescence signal could be observed in vivo. The development of tumor could be found in the left lateral lobe through ICG surgical navigation system. (2) Even though BPL and PVL could both lead to the AHC (Atropy/hypertrophy complex) of the liver, BPL was faster and more effective. The intrahepatic bile acid of BPL group reached the peak value on the first day after the operation, and declined rapidly afterwards. The intrahepatic bile acid of PVL group reached the peak value on the third day after the operation and then declined slowly. Bile acid reduced to normal level on 14 days after the ligation. (3)The level of expression of PCNA and the ki-67 count in BPL group peaked in a shorter time compared with those in the PVL group. (4)Compared with Sham group, both BPL and PVL group enhance the growth of tumor at day7 after operation. However, the tumor in the BPL group is larger than that in the PVL group at day 14 after operation. Pulmonary metastasis at day 14 after BPL could be observed, and the metastasis rate is 100%(4/4).Conclusion (1) The orthotopic implantation of Luc-McA-RH777 cell could form tumors in Buffalo rats, and their growth could be observed by Luciferase fluorescent imaging system and ICG surgical navigation system. But Luciferase fluorescent imaging was more sensitive than ICG surgical navigation system. (2) Compared with PVL, BPL is more effective in inducing hypertrophy of the non-ligated liver lobes. (3) The combined ligation of the portal vein and bile duct can accelerate the development and metastasis of liver cancer. |
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