Construction Of TACE Model And Microwave Ablation Model Of Murine In Orthotopic Hepatocellular Carcinoma

ObjectiveTo establish a caudal artery-route transcatheter arterial chemoembolization(TACE)model in diethylnitrosamine-induced(DEN-induced)rat with orthotopic Hepatocellular carcinoma(HCC)and to evaluate its feasibility,efficiency and safety.MethodsRats were treated with 0.01%N-diethylnitrosamine(DEN)solution for 12 weeks to induced spontaneous orthotopic HCC.At the end of 12 weeks,rats were scanned by magnetic resonance imaging(MRI)and the ones with maximum tumor diameter beyond 5mm were included.To conduct TACE,the microcatheter was inserted through caudal artery and super-selectively intubated into the proper hepatic artery of HCC rat.Then the mixture of lipiodol and doxorubicin were injected into the tumor through microcatheter.After TACE,intra-tumoral lipiodol deposition,animal behavior,hepatic function indicated by alanine aminotransferase(AST),aspartate aminotransferase(ALT)and total bilirubin(TBIL),as well as renal function indicated by creatinine(CRE)were evaluated.Moreover,the liver,kidney,lung and spleen tissues were collected for histopathological assessment at the 7th day after TACE.ResultsThe TACE model was successfully established with an average time of 6.9±1.4 minutes for microcatheter insertion.Obvious intrahepatic tumor staining was revealed by intraoperative arteriography and dense lipiodol deposits within the tumor were confirmed by postoperative CT scans.The histopathological results verified extensive necrosis in the tumor center and showed no ectopic emboli in the lungs,kidney and spleen.No severe complications in rats were observed as evidenced by mild insults on hepatic function and by the absence of abnormal animal behaviors.ConclusionA caudal artery-route TACE model is feasible,efficient and safe In DEN-induced rat with orthotopic HCC.ObjectiveTo construct hepatocellular carcinoma(HCC)cells lines that can mimic Sub-lethal heat shock(HS)in vitro and orthotopic HCC mouse model that received incomplete thermal ablation.MethodsThe suspension H22 cells were grown in 6-well plates,and the medium was replaced with fresh complete medium 8h before heat treatment.The plate was placed in a constant temperature water bath at 37℃、42℃、47℃、50℃ for 15 min.After the heat treatment was completed,the cells were maintained in a 37℃ incubator until detection.The phenotype of epithelial-mesenchymal transition(EMT)was detected by Western blot 48 hours later,and the cell viability was detected by Cell Counting Kit-8(CCK-8)kit.The liver of mice was exposed by laparotomy along the abdomen midline and H22 cells(5×106)were injected through the hepatic capsule to conduct orthotopic HCC model.When the longest diameter of the tumor reached approximately 8 mm,mice were randomly divided into complete ablation group(cMWA),incomplete ablation group(iMWA)and Control group treated with sham operation.A micro-MWA antenna was inserted percutaneously along the long axis until the tip of the antenna reached the contralateral edge of the tumor.MRI was performed for tumor measurement and liver specimens were collected for histopathological assessment at day 14 after microwave ablation.Results14 days after the injection of H22 cells,the tumors reached average tumor diameter of 9.32±0.83mm which was predetermined to be suitable for microwave ablation.14 days after microwave ablation,the average tumor diameter reached 12.93±1.51mm in iMWA group,which were significantly bigger than preoperative period.In addition,immunofluorescence staining showed a large number of infiltrated neutrophils in the residual tumor of iMWA group.Western blot was used to observe the change in EMTphenotype,results showed that Vimentin and a-SMA was up-regulated and E-cadherin was down-regulated.ConclusionHCC cells lines that can mimic Sub-lethal heat shock(HS)in vitro and orthotopic HCC mouse model that received incomplete thermal ablation were successfully constructed.There was substantial staining for neutrophils infiltration after incomplete ablation.Incomplete ablation may allow HCC cells to become more prone to migrate and to become more invasive.