To evaluate the application value and feasibility of the updated method of trans-gastroduodenal reverse catheterization of hepatic artery in a rat model of Hepatocellular Carcinoma (HCC) via orthotopic implantation of histologically intact tissue. 120 Wistar rats were randomly divided into 4 groups (30 rats/group). Groups A and B were preliminary experimental groups, in which rats were respectively treated with modified and traditional methods of hepatic arterial cannula. HCC model was established in rats of groups C and D by puncture transplantation, where rats were also treated with hepatic artery angiography and interventional embolization by modified and traditional methods, respectively. Comprehensive comparison and analysis were conducted on the success rates of cannula and HCC model establishment, variations of tumor volume, death rates and causes of rats throughout the surgery. The success rate of cannula was 90% in group A and 63.3% in group B. Obviously, the success rate of group A was higher than that of group B (χ2=4.565, P=0.033). In addition, the success rate of cannula was up to 93.3% in group C and 70% in group D. Statistically, the success rate of group C was apparently higher than that of group D (χ2=4.007, P=0.045). During the whole experiments, some rats mainly died from vascular damage, liver injury, and anesthesia accident. However, the success rate of tumor implantation reached up to 100% in rats of groups C and D. The mortality of group D was clearly higher than that of group C during the initial 10 days postoperatively (42.86% versus 10.71%, χ2=5.079, P=0.024), while interventional embolization treatment had no statistical influence on average tumor volumes. The modified application of trans-gastroduodenal artery reverse catheterization of hepatic artery in rats took the advantages of easier operation, higher success rate, lower postoperative mortality, and lesser complications. It was worthy to be popularized in clinical trials and deserved a deeper exploration.