Objective To observe the effects of extracellular-signal regulated kinase (ERK) 1/2 inhibitor U0126 on hepatoma carcinoma cell proliferation and apoptosis. Methods Hepatoma SMMC-7721 cell strain was divided into blank control group and different concentrations of U0126 groups. The proliferation inhibition was measured by MTT assay. FCM was used to analyze the cell cycle distribution and apoptosis. Results U0126 obviously inhibited cell proliferation, induced cell apoptosis and G0/G1 phase cell cycle arrest. There were significant differences between control group and different concentrations of U0126 groups on cell proliferation and apoptosis （P＜0.05， P＜0.01）. Conclusion Blocking ERK1/2 pathway may be an important treatment strategy for liver cancer.
Objective To establish the model of hepatic VX2 tumor in rabbits and to offer the experimental evidences for the application of the model. Methods The hepatoma model was reproduced with VX2 cell lines in rabbits. The method to reproduce the model was improved. The changes of liver function (ALT, AST and TB) were determined at a different phase. Tumor’s growth and metastases, pathological changes, images and spontaneous survival time of the animal were observed. Results The tumors could grow up to 1.5-2.0 cm in diameter in 3 weeks after implanting. The successful rate of implantation was 100%. Nodular enhanced echo was found in the liver by color ultrasound. CT scans showed the low density foci in liver, while enhanced CT scans demonstrated asymmetrical intensification in the foci. Macroscopic observation showed that the tumors were grayish white in color and felt harder, necrotic foci was present in the center of tumor. Observation with light microscope showed that the tumor cells’ nucleoplasm proportion was great, tumor cells arranged irregularly, and the tumors displayed invasive growth and no obvious envelope around them. Animals’ spontaneous survival time was 40-53 days. The cause for their death was multiple system organ failure. Conclusion In pathological morphology, pathological process and prognosis, the hepaticVX2 tumors in rabbits are similar to human hepatocarcinoma. It has such characteristics as easy reproduction, short growth period, high success rate, high stability and so on. The model is an ideal hepatoma model in animals.
For research the relationship between the expression of vascular endothelial growth factor (VEGF) and the cell proliferation. The expression of VEGF was evaluated while the cell cycle of hepatoma cell line Hep G2, which was synchronized at G0 phase with serum deprivation, and reinitiated with TPA and blocked with antisense oligonucleotides of c-jun. Results: Hep G2 cell did not express VEGF at G0 phase. TPA could induce the expression of VEGF as well as initiation of cell cycle. The antisense oligonucleotides of c-jun could prohibit the expression of VEGF and arrest the cell cycle at G0 phase. Conclusion: The fact that the expression of VEGF accompanies the initiation of cell cycle suggests that they be regulated by the same singnal pathway, the expression of VEGF may be a marker indicating the proliferation of hepatoma cells.
This paper reports twelve patients underwent repeat hepatic resection because of the recurrence of hepatocellular carcinomas after primary resection. The indication of reoperation, selection of incision, difficults encountered in the operation and the treatment after operation are discussed. The authors believe that the second operation is technically more difficult than the first one, some troubles my be happened during the operation and put forward some ways to deal with this situations.
Four pigs underwent the hepatic arterial infusion with 32P glass microsphere (32PGM) and pigs were killed in 15th, 30th and 90th days separately. Pathological study showed that in early stage there were many small necrotic areas scattered along the hepatic arterioles. Three months later, these necrosis were gradually absorbed and replaced by regenerating hepatic cells. Tumor-inhibition experiment was performed in 40 Bal B/C mice bearing H22 hepatoma. Intratumoral injection of 0.2ml of 32PGM/glycerine suspension (group A, n=20) or 0.2ml of blank glass microsphere/glycerine suspension (group B, n=20) were performed. The average survival time in group A and group B was 24.8 and 11.8 days respectively. Five mice in group A were alive beyond 40 days after treatment, disappearance of tumor was found in two of them. This experiment demonstrates that 32PGM is effective for treatment of experiment hepatoma. The damage to hepatic tissue after infusion is associated with the irregular distribution of microsphere, and this lesion can completely recover within three months.
ObjectiveTo explore the safety and feasibility of contrast enhanced intraoperative ultrasonographyguided percutaneous radiofrequency ablation with artificial hydrothorax to hepatocellular carcinoma in the hepatic dome. MethodsThe clinical data of nine patients with hepatocellular carcinoma in the hepatic dome underwent ultrasonographyguided percutaneous radiofrequnecy ablation with artificial hydrothorax from January 2008 to June 2009 at Department of Hepatobiliopancreatic Surgery of West China Hospital were retrospectively analyzed. The perioperative results and recurrence of tumor were also analyzed. ResultsAll of nine patients with twelve tumors received successfully radiofrequency ablation with artificial hydrothorax of (2 444±464) ml (2 000-3 000 ml). The ablation time was 12-24 min (median 12 min), with an average of (15±5) min for each tumor. No hemothorax, pneumothorax, and death occurred during operation. One patient had ascites of 2 000 ml after ablation due to hypoalbuminenia, and ascites disappeared by infusion of abumin on 4 d after operation. The total volume of pleural drainage was 250-1 420 ml, with an average of (717±372) ml for each patient, and the drainage tube was withdrawn on 3-5 d after operation. The followup time was 7-23 months (mean 15 months). Tumor recurrence was found in three patients on 5, 6, and 7 months after operation, respectively. Of them, two patients were in stable disease stage after interventional and conservative therapy, respectively, and one case recurred at six months after operation and died of hypertensive heart disease and hepatic function deterioration at sixteen months after operation. The rest patients survived and no recurrence and metastasis was observed during the follow-up period.ConclusionThe technique of percutaneous radiofrequency ablation with artificial hydrothorax increases the feasibility of the minimal invasive treatment for hepatoma, which can be applied to hepatocellular carcinoma in the hepatic dome with high safety and clinical application value.
【Abstract】Objective To investigate the apoptosis induced by TGF-β1 in human hepatic carcinoma cell lines and its relationship with p53 gene and Smad. Methods Three human hepatic carcinoma cell lines which involving in various status of the p53 gene were used in this study. TGF-β1-induced apoptosis in hepatic carcinoma cell lines was measured by the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. To study the mechanism of TGF-β1-induced apoptosis, these cell lines were transfected with a TGF-β1-inducible luciferase reporter plasmid containing Smad 4 binding elements (SBE) and luciferase gene using Lipofectamine 2000, then treated with TGF-β1, relative luciferase activity was assayed. Results Of three cell lines studied with TUNEL assay, TGF-β1 induced apoptosis was observed in HepG2 cells (wild type p53). Huh-7 (mutant p53) and Hep3B (deleted p53) cell lines showed less apoptosis. Luciferase activity assay indicated that the response to TGF-β1 induction in HepG2 cells was increased dramatically but was not significant in Huh-7 and Hep3B cell lines. Conclusion HepG2 cells seem to be highly susceptible to TGF-β1-induced apoptosis compared with Hep3B and Huh7 cell lines. Smad 4 is a central mediator of the TGF-β1 signal transduction pathway.
ObjectiveTo investigate the effects of celastrol on the growth and apoptosis of huamn hepatoma SMMC-7721 cells, and investigate its preliminary action mechansim. MethodsSMMC-7721 cells were cultured in vitro, CCK-8 assay and Annexin V-FITC/PI staining method were conducted to investigate the effects of celastrol on the growth and apoptosis of huamn hepatoma SMMC-7721 cells after the cells were treated with drugs, and then the Caspase-3 activity and NF-κB protein expression were determined by Caspase-3 activity determination kit and Western blot. Huamn hepatoma SMMC-7721 cells transplantation tumor models in nude mice were established and the effect of celastrol on the growth of transplantation tumor were observed. ResultsCelastrol could inhibit the SMMC-7721 cells growth in a dose and time dependent manner. Annexin-V/PI staining showed that SMMC-7721 cells were induced to death with the concentration increasing of celastrol. Caspase-3 activity was measured after treatment with celastrol and the results indicated that the activity of caspase-3 was significantly enhanced. Western blot experiments showed that the expression of NF-κB protein decreased in a time-dependent manner after treatment with celastrol. Celastrol could inhibit SMMC-7721 cells transplantation tumor growth in nude mice. ConclusionsCelastrol could inhibit the proliferation of human hepatoma SMMC-7721 cells and induces apoptosis, and inhibit SMMC-7721 cell transplantation tumor growth in nude mice. Celastrol induce apoptosis of SMMC-7721 cells might through activating Caspase-3 pathway and NF-κB pathway.