Hepatocellular carcinoma(HCC)is a major cause of cancer mortality worldwide,and liver transplantation (LT)has the potential to improve the survival for patients with HCC.However,tumor recurrence after LT remains the main obstacles for long-term survival.Selection of the ideal recipients based on Milan criteria or Shanghai Fudan criteria is a key point to reduce the incidence of recurrence.C,enomics and proteomics combined with tumor specific tumor markers detection are helpful to screen out the recipients.The norms of rumor-free operation should be strictly followed intraoperatively.Preventive chemotherapy and evaluation of immune function should be considered postoperatively to reduce the risks of tumor recurrence and metastasis.

Objective To investigate the effects of inhibiting epithelial mesenchymal transition (EMT) on metastastic potential of hepatocellular carcinoma (HCC) enhanced by hepatic arterial occlusion in mice. Methods Using a metastatic human HCC orthotopic nude mice model (MHCC97),the effects of hepatic artery ligation (HAL) alone,combination of HAL and phosphatidylinositol 3-kinase (PI3K)inhibitor LY294002,or combination of HAL and interferon-α (IFN-α) on the growth of planted human HCC cells and pulmonary metastasis were evaluated,respectively.The cells and tumor tissues specimens were analyzed through expression of Akt,p-Akt,E-cadhein,N-cadherin and Twist. Results HAL inhibits tumor growth (2002.97 ± 331.28 ) mm3 vs.( 3921.23 ± 786.21 ) mm3,t =4.052,P ＜ 0.01 ),while promoting pulmonary metastatsis ( 10/12 vs.4/12,P ＜ 0.05).HAL combined with LY294002 represses significantly enhanced pulmonary metastasis rate by HAL alone (0/6 vs.6/6,P ＜ 0.01 ).Moderate-dose IFN-α (7.5 × 106 U/kg) combined with HAL failed to further reduce tumor volume compared with that of HAL alone,but inhibited markedly pulmonary metastasis (2/6 vs.6/6,P ＜ 0.05 ).The augmented level of N-cadherin and Twist in tumor tissues due to HAL reduced by LY294002 or 7.5 × 106 U/kg IFN-α.The arrest of EMT by LY294002 or IFN-α in HAL-treated xenografts was further demonstrated by the in vitro response of hypoxic cells to both agents. Conclusions Inhibition of EMT in HCC cells could repress enhanced metastastic potential due to hepatic arterial occlusion.

Post-transplant tumor recurrence and metastasis remain the main obstacles for long-term survival after liver transplantation (LT) for hepatocellular carcinoma (HCC). Measures to explore the HCC biological characteristics and the relationship between post-transplant immuno-suppression and tumor recurrence, to determine precisely the prognostic factors associated with post-transplant recurrence, to intervene effectively for those with high risk of recurrence, and to use individualized multimodality treatment for recurrence and metastasis may improve the therapeutic results of LT for HCC.

Objective To study the effect of low molecular weight heparin on the growth and metastasis of hepatocarcinoma in vivo Methods Metastatic model simulating human hepatocarcinoma was established by orthotopic implantation of the tumor tissue into nude mice Animals were randomly divided and submitted to different treatment groups: control group treated with nornal saline, chemotherapy group with 5 FU and CDDP i p , Fragmin group (low molecular heparin), combination group After 5 weeks of treatment, tumor volume、 microvessel density (MVD)、 VEGF smoothmuscle actin (SMA) and metastasis in Fragmin group significantly decreased and tumor inhibitive rate increased ( P

Objective To investigate the effects of hypoxia on malignant phenotype of a hepatocellular carcinoma(HCC)cell line and its molecular basis.Methods Human HCC cell line with highly metastatic potential(MHCC97H)was grown under hypoxia(induced by 100 μmol/L CoCl_2)and normoxic conditions respectively.To observe the effects of hypoxia on MHCC97H cells was observed,the tumorigenicity was carried out by soft agar cloning method in vitro and inoculation in nude mice in vivo;Invasive and metastatic potential were measured.Experiments with and/or without Matrigel in vitro and a metastatic human HCC orthotopic nude mice model by HAL in vivo.To clarify the molecular background,the proliferation of cells were analyzed by MTT assay and the cell cycle was detected by flow cytometry;The expression of embryonic stem cell(ES)-like genes(Oct4,Nanog and Sox2)were detected by real-time RT-PCR;CD90~+ and CD133~+ subpopulation from MHCC97H cells were isolated by flow cytometry and the expressions of CD90 and CD133 of MHCC97H cells were analyzed by immunofluorescence.Results Hypoxia increased tumor migration(t=2.792,P=0.023),invasiveness(t=7.624,P＜0.0001)and clone forming ability(t=3.292,P=0.011)of MHCC97H cells in vitro,and promoted tumorigenesis(x~2=8.571,P=0.015)and pulmonary metastasis(x~2=5.507,P=0.031)in vivo.The proliferation of MHCC97H cells arrest under hypoxic conditions accompanied with increased proportion of cells in G1 phase,the expressions of Oct4,Nanog,Sox2 significantly increased in response to hypoxia,but the fluorescence intensity and number of CD90~+ and CD133~+ MHCC97H cells decreased or unchanged.Conclusion Hypoxia increases aggressiveness of MHCC97H cells,which was correlated with the acquisition of embryonic stem cell-like characteristics.

Complete resection of liver cancer is the main approach for achieving radical resection,and sufficient remnant liver is essential for avoiding hepatic failure after operation.With the aim of increasing remnant liver volume,a new two-stage technique,associating liver partition and portal vein ligation for staged hepatectomy (ALPPS),recently has been developed.In this article,the initial experience with 1 case of hepatocellular carcinoma who underwent ALPPS at the Zhongshan Hospital in April 2013 was reported.In the first stage,the right portal vein branch was ligated and subsequently the liver parenchyma was dissected along the falciform ligament to isolate the segment Ⅳ and the left lateral lobe.On postoperative day 7,the remnant liver volume was increased from 291 ml to 579 ml,and on postoperative day 8,the second stage operation was performed.During the second stage,the extended right lobe was removed.ALPPS induces a great and fast hypertrophy of the remnant liver,and R0 resection could be performed on patients which was considered unresectable because of small remnant liver volume,without severe postoperative liver failure and has a low mortality.

Objective To investigate the role and mechanism of TMPRSS4 in radiation induced metastasis of hepatocellular carcinoma (HCC).Methods Metastatic model of human HCC was established by orthotopic implantation of histologically intact human HCC tissue into the liver of nude mice.Mice bearing xenografts in liver were killed after radiation and the residual tumors were resected and reimplanted into the liver of normal nude mice.At the end of sixth week,the mice were killed and the histopathological features,tumor volume,intrahepatic and lung metastasis were evaluated.Expression of epithelial-mesenchymal transition (EMT) related genes including N-cadherin,Vimentin,SIP1 and TMPRSS4 were measured by Western blotting and RT-PCR.Results The tumor volume and frequency of lung metastasis of control group was 2.25±0.52 cm3 and 66.7％,respectively.Compared to control group,tumor diameter (1.61±0.51 cm3,P＜0.05) and lung metastasis (12.5％,P＜0.05) were significantly inhibited 2 days after radiation.Whereas,30 days after radiation,tumor growth recovered (2.60±0.61 cm3,P＞0.05) and lung metastasis was enhanced (100％,P＜0.05).There were no intrahepatic metastasis in the control group and in the group of reimplantation of HCC 2 days after radiation,while the tumors from those 30 days after radiation showed enhanced intrahepatic metastasis (18 ± 8.05,P＜ 0.01 ),with overexpression of SIP1,N-cadherin,Vimentin and TMPRSS4,and reduced expression of E-cadherin.Conclusion The metastasis potential of residual HCC after radiation was first inhibited and then promoted.Overexpression of TMPRSS4 plays a critical role in radiation induced long-term metastasis of HCC by facilitating EMT.

ObjectiveTo study the role and mechanism of low-dose aspirin with IFN-α in inhibiting growth and metastasis of hepatocellular carcinoma (HCC).MethodsMHCC97L cells were cultured and a metastatic model of human HCC was established by orthotopic implantation of histologically intact human HCC tissue into the liver of nude (nu/nu) mice.After administration of different doses of Aspirin and IFN-α for 40 days,the mice bearing xenografts in liver were killed,and the tumor volume and lung metastasis were evaluated.Cell proliferation and MMP-2 activity were measured by MTT and gelatin zymography,respectively.The expressions of VEGF and MMP-2 were measured by western blot and ELISA.ResultsCompared to the control group,there were no significant differences in the high-dose Aspirin [45 mg/(kg · d)] treated group regarding tumor volume [(1.89 ±0.88) cm3 vs (3.12±0.85) cm3,P＞0.05] and incidence of lung metastases (58.3％ vs 66.7％,P＞0.05),but the tumor volume and incidence of lung metastasis were significantly inhibited in the highdose IFN-α group [1.5 × 107/(kg · d)],the high-dose IFN-α combined with high-dose Aspirin group,and the low-dose IFN-α [7.5 × 106 / (kg · d) ] combined with low-dose Aspirin [15 mg/(kg · d] group (P＜0.05).2 mmol/L Aspirin did not inhibit the proliferation of MHCC97 cells (P＞0.05),but inhibited the activities and expressions of MMP-2 and VEGF.Low-dose IFN-α combined with low-dose Aspirin significantly decreased the expressions of MMP-2 and VEGF in nude mice (P＜0.05).ConclusionLow-dose Aspirin combined with low-dose IFN-α significantly inhibited the growth and metastasis of HCC through suppressing the expressions of MMP-2 and VEGF.

Objective To summarize the techniques of anatomical liver resection for the treatment of hepatocellular carcinoma(HCC).Methods The clinical data of 125 patients with solitary HCC who underwent anatomical liver resection at the Zhongshan Hospital from January 2005 to December 2006 were retrospectively analysed.The inflow and outflow of hepatic segments to be resected were selectively clamped,then the main branches of portal vein and hepatic artery were ligated,and the ischemic hepatic segments were resected en bloc.Kelly forceps were used to crash and clamp the liver cut surface.The stumps of left and right hepatic ducts were continuously sutured with Prolene sutures.For tumors with the size above 10 cm in diameter,hepatectomy with anterior approach and liver hanging maneuver were adopted.Bile leakage was checked by injecting methylene blue or covering a gauze on the liver cut surface.Results The mean blood loss of all patients was 250 ml(100-6000 ml),and 32 of them needed blood transfusion.The morbidity was 23%(29/125).No patient died within 30 days after the operation,and 6%(5/83)of patients were found with residual tumor by postoperative arteriography.Conclusion Anatomical liver resection may improve the safety of operation,prevent the injury of great vessels and thus improve the efficacy.

Objective To summarize the clinical experienee in surgical treatment for hepatocellular carcinoma (HCC). Methods The clinical data of 7566 HCC patients who had been admitted to Research Institute of Liver Cancer of Fudan University from January 1988 to Deeember 2007 were retrospectively analyzed. The overall survival and recurrence free survival (RFS) rates were eaeulated with Kaplan-Meier survival curve. All the data were analyzed using Log-rank test and Cox regression model. Results The 3-, 5-, 10-year overall survival and RFS rates of 7164 patients with HCC resection were 56.29%, 41.76%, 26.70%, and 63.92%, 56.12%, 42.97%, respectively, and the perioperative mortality was 1.54%. The 5- and 10-year overall survival rates of patients with small HCC (diameter<5 era) were 58.20% and 38.47%, which were significantly higher than 31.42% and 20.43% of patients with large HCC (diameter >5 cm) (X2 =535. 568, P <0.01). The 5-year overall survival rotes of HCC patients with resection after down-staging (n = 110), re-resection after recurrence (n = 515), and tumor thrombus in portal vein (n = 168) were 51.26%, 67.28% and 26.81%, respectively; nd the 5-year DFS rotes were 77.44%, 13.01% (calculated from the first operation) and 34.90%, respectively. The 3- and 5-year overall survival and DFS rates of 402 patients who had undergone liver transplantation were 60.81%, 55.63% and 64.47%, 58.52%. The independent prognostic factors influencing the overall survival and DFS rates were the size, number and differentiation of HCC and intrahepatic vessel invasion (X2 = 200.539, 27. 536, 96.964,216. 156, P <0.01). Conclusions Early screening, improved safety of surgery, combined therapy and breakthrough in the reseaeh of preventing HCC metastasis and reeurrenee will significantly improve the treatment outcome of HCC.