ObjectiveTo identify the risk factors of postoperative recurrence and survival for patients with hepatocellular carcinoma within Milan criteria following liver resection. MethodsData of 267 patients with hepatocellular carcinoma within Milan criteria who received liver resection between 2007 and 2013 in our hospital were retrospectively analyzed. ResultsAmong the 267 patients, 123 patients suffered from recurrence and 51 patients died. The mean time to recurrence were (16.9±14.5) months (2.7-75.1 months), whereas the mean time to death were (27.5±16.4) months (6.1-75.4 months). The recurrence-free survival rates in 1-, 3-, and 5-year after operation was 76.8%, 56.3%, and 47.6%, respectively; whereas the overall survival rates in 1-, 3-, and 5-year after operation was 96.6%, 82.5%, and 74.5%, respectively. Multivariate analyses suggested the tumor differentiation, microvascular invasion, and multiple tumors were independent risk factors for postoperative recurrence; whereas the tumor differentiation, positive preoperative HBV-DNA load, and preoperative neutrophil-to-lymphocyte ratio adversely influenced the postoperative survival. ConclusionsFor patients with hepatocellular carcinoma within Milan criteria after liver resection, the tumor differentiation, microvascular invasion, and multiple tumors contribute to postoperative recurrence; whereas the tumor differentiation, positive preoperative HBV-DNA load, and preoperative neutrophil-to-lymphocyte ratio adversely influence the postoperative survival.
Objective To investigate immunological therapeutic effect and safety of dendritic cells (DCs) combined with heat shock protein 70 (HSP70)-peptide complex (PC) derived from autogeneic hepatoma tissue. Methods Thirty patients with hepatocellular carcinoma from February 2010 to February 2015 in the Gaochun People’s Hospital of Nanjing and The Third Affiliated Hospital of Nantong University were studied, and subsequently were divided into an immunotherapy group (treated with HSP70-PC/DCs vaccine,n=15) and a chemotherapy group (n=15) according to the prescribed postoperative treatment methods. The levels of T lymphocyte subtypes were assayed by FACS. The toxicity adverse reactions, alpha-fetoprotein (AFP), CA19-9, hepatic tumor recurrence rate, survival rate, and KPS of two groups patients were evaluated and compared between these two groups. Results ① The values of CD3+, CD4+, CD4+/CD8+, and CD3CD56 had no significant differences between the immunotherapy group and the chemotherapy group before treatment (P>0.05), which in the immunotherapy group were significantly higher than those in the chemotherapy group after treatment (P<0.05), and which were significantly higher in the immunotherapy group after treatment as compared with the levels before treatment (P<0.05), and which had no significant differences in the chemotherapy group between after treatment and before treatment (P>0.05). ② Before treatment, the levels of AFP and CA19-9 had no significant differences between the immunotherapy group and the chemotherapy group (P>0.05), which in the immunotherapy group were significantly lower than those in the chemotherapy group after treatment (P<0.05). In the immunotherapy group, the levels of AFP and CA19-9 after treatment were significantly lower than those before treatment (t=2.564，P=0.021；t=2.011，P=0.041), which in the chemotherapy group before treatment were decreased as compared with the levels before treatment (t=2.221,P=0.036;t=2.487,P=0.066). ③ The patients treated with the HSP-PC/DCs vaccines was well tolerated and no obvious toxicity was appeared. ④ All the patients were followed up 5–19 months with median follow-up time of 9 months. The median survival time was 560 d and 436 d in the immunotherapy group and the chemotherapy group, respectively. After treatment, KPS score was significantly higher and recurrence rate was significantly lower in the immunotherapy group as compared with the chemotherapy group (P<0.05). The total survival had no significant difference between the immunotherapy group and the chemotherapy group (P>0.05). Conclusions The preliminary results of limited cases in this study show that HSP70-PC/DC vaccination is safe and effective in treatment of hepatocellular carcinoma, the pulsed DCs are effective in activating specific T-cell responses against hepatocellular carcinoma cells. HSP70-PC/DC vaccine might improve immunity and prevent postoperative recurrence of hepatocellular carcinoma.
Objective To investigate whether the growth of the human experimental hepatocellular carcinoma (HCC) can be suppressed by the antibody against vascular endothelial growth factor (VEGF). MethodsThe monoclonal antiVEGF antibody was injected to nude mice nearby the xenograft tumour foculs of the human SMMC7721 HCC. The changes of the tumour size were measured at different times. The intratumoural microvessels were showed by immunohistochemical staining of CD31 antigen; the apoptotic cells in the tumour tissues were detected in situ by terminal deoxynucleotidyl transferasemediated dUTPbiotin nick end labeling (TUNEL) assay. ResultsIn the first and second week after finishing the injection procedure, the tumour sizes were compared as the length (mm) multiplying width (mm) between the two groups, the tumour sizes as the test group vs the control group were (26.46±19.81) mm2 vs (105.77±17.40) mm2 (P＜0.001) and (45.20±23.02) mm2 vs (150.77±77.41) mm2 (P＜0.05), respectively. After 2 weeks the intratumoural microvessel density (iMVD) and the apoptotic index (AI) were compared between two groups with iMVD being (2 311±120)/mm2 vs (3 900±328)/mm2(P＜0.001 ) and AI being (15.31% vs 6.83%), P＜0.005. Conclusion The antiVEGF antibody can suppress the xenograft tumour growth of the human SMMC7721 HCC by antiangiogenesis.In fact, its antitumour effect is produced by elevating the incidence of apoptosis in tumour tissues.
Objective To introduce the possible effects and significances of angiogenesis and antiangiogenic in the development and treatment of hepatocellular carcinoma (HCC). Methods Recently relevant literatures were reviewed. Results Angiogenesis played a significant role in the development and therapy of HCC, and the development and metastasis of HCC could be effectively suppressed by antiangiogenic therapy. This might provide a new approach for the treatment of HCC. Conclusion Comprehending the molecular mechanism of angiogenesis and applying antiangiogenic therapy will contribute a lot for the prevention and treatment of HCC.
Objective To construct the recombinant of hepatocellular carcinoma-targeting adenovirus containing r-Caspase-3 gene and provide the gene therapic strategy for hepatocellular carcinoma. Methods The pAdTrack-EAFP-PALB was constructed and the r-Caspase-3 gene was subcloned into the vector. The linearized shuttle plasmid was homogenously recombined with AdEasy-1 in BJ5183 cells. The candidate clone was analyzed by restriction endonuclease digestion and sequencing, and then pAdEasy-EAFP-PALB/r-Caspase-3 vector was digested with PacⅠand transfected into AD293 cells for packaging and amplifying, recombinant virus was constructed successfully. Infection titer and efficiency of recombinant virus were monitored by green fluorescent protein (GFP) expression. The expression of r-Caspase-3 in infected HepG2 cells was detected by RT-PCR and Western blot. The apoptosis of HepG2 cells was detected by SRB dyeing method. Results Shuttle vector pAdTrack-EAFP-PALB/r-Caspase-3 was correct after identification by restriction endonuclease analysis and sequencing. By PCR and PacⅠ restriction endonuclease analysis， the homologous recombinant of pAdEasy-EAFP-PALB/r-Caspase-3 was successful. The expression of GFP was observed when linearized pAdEasy-EAFP-PALB/r-Caspase-3 was transfected into AD293 cells. AD293 cells could be infected repeatedly by recombinant adenovirus. The expression of r-Caspase-3 gene on HepG2 cells was detected by RT-PCR and Western-blot methods respectively, which confirmed that the Ad-EAFP-PALB/r-Caspase-3 was constructed successfully. The specificity of Ad-EAFP-PALB/r-caspase-3 which targeting induced hepatocellular carcinoma cells was founded by SRB dyeing test. Conclusion The Recombinant of hepatocellular carcinoma-targeting adenovirus containing r-Caspase-3 gene was constructed successfully and which established the foundation of r-Caspase-3 gene therapy in future research to hepatocellular carcinoma.
ObjectiveTo analyse the outcomes of patients with Child-Pugh A class cirrhosis and a single hepatocellular carcinoma (HCC) up to 5 cm in diameter who underwent liver transplantation versus resection. MethodsDuring 2007 to 2011, 263 Child-Pugh A class cirrhotic patients with a single HCC up to 5 cm in diameter either underwent liver resection (n=227) or received liver transplantation (n=36) in our centre. Patients and tumour characteristics and outcomes were analysed. ResultsThe 1-, 3-, and 5-year recurrence-free survival rates of patients who received liver transplantation and liver resection were 91.7%, 85.3%, 81.0% and 80.6%, 59.8%, 50.8%, respectively (P=0.003). The 1-, 3-, and 5-year overall survival rates of patients who underwent liver transplantation were 100%, 87.5%, and 83.1% versus 96.9%, 83.8%, and 76.1% for patients received liver resection (P=0.391). The 1-, 3-, and 5-year recurrence-free survival rates for patients with a diameter of HCC < 3 cm underwent liver transplantation were 92.3%, 92.3%, and 92.3% versus 80.2%, 62.5%, and 50.5% for live resection group (P=0.019). The 1-, 3-, and 5-year overall survival rates for patients with a diameter of HCC < 3 cm underwent liver transplantation and liver resection were 100%, 91.7%, 91.7% and 97.7%, 87.5%, 79.5%, respectively (P=0.470). ConclusionsAlthough more recurrences are observed in Child A class cirrhotic patients with a single HCC up to 5 cm in diameter after liver resection, but overall survival rates for patients with a single HCC up to 5 cm in diameter are similar after liver resection and transplantation.
Objective To explore the clinical value of gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid （Gd-EOB-DTPA） enhanced magnetic resonance （MR） imaging for cirrhosis-related nodules. Methods Nineteen patients who were suspected cirrhosis with lesions of liver were prospectively included for Gd-EOB-DTPA enhanced MR imaging test between Nov. 2011 and Jan. 2013. The hepatobiliary phase （HBP） images were taken in 20 minutes after agents’ injection. The images were diagnosed independently in two groups： group A， including the plain phase and dynamic phase images； group B， including plain phase， dynamic phase， and HBP phase images. The signal intensity （SI） of lesions in HBP images， background liver SI， and background noise standard deviation were measured by using a circular region of interest， then the lesion signal to noise ratio （SNR） and contrast signal to noise ratio （CNR） were calculated. Results Nineteen patients had 25 tumors in all， including 18 hepatocelluar carcinoma （HCC） and 7 regenerative nodule （RN） or dysplastic nodule （DN）， with the diameter ranged from 0.6 cm to 3.2 cm （average 1.3 cm） . Sixteen HCC manifested hypo SI relative to the normal liver， while 2 HCC manifested hyper SI at HBP. Five HCC had cystic necrosis with the necrotic area， and there were no enhancement in artery phase， while performed flocculent enhancement at HBP. Six RN or DN showed hyper SI while another 1 showed iso SI to background liver at HBP. The diagnostic accuracy rates of group A and group B were 80.0% （20/25） and 92.0% （23/25）. SNR of RN or DN at HBP was 132.90±17.21， and of HCC was 114.35±19.27， while the CNR of RN or DN was 19.47±8.20， and of HCC was 112.15±33.52. Conclusion Gd-EOB-DTPA enhanced MR imaging can improve the diagnosis capacity of cirrhosis-related nodules， so as to develop more accurate and reasonable treatment options.
ObjectiveTo evaluate the prognostic value of the easy albumin-bilirubin (EZ-ALBI) score for postoperative complications and long-term prognosis of hepatocellular carcinoma (HCC) patients. MethodsThe data on consecutive 1 822 HCC patients who underwent hepatectomy were obtained and retrospectively analyzed from five medical centers, including West China Hospital, Sichuan Provincial People’s Hospital, The First People’s Hospital of Neijiang City, The Second People’s Hospital of Yibin City, and People’s Hospital of Leshan City. Non-conditional logistic and Cox proportional hazards regression were used to evaluate the aspect on the postoperative complications and long-term prognosis. ResultsThe patients in EZ-ALBI grade 2 had higher incidences of severe complication (Clavein-Dindo classification>2, P=0.001), post-hepatectomy liver failure (P=0.040), length of stay>10 d (P<0.001), perioperative transfusion (P<0.001), and 90 d mortality (P<0.001). The 1-, 3- and 5-year cumulative survival rates in EZ-ALBI grade 1 group were 85.5%, 67.0%, and 58.7% while in EZ-ALBI grade 2 group were 72.7%, 51.1%, and 39.8%. Multivariate Cox proportional hazards regression manifested that patients in EZ-ALBI grade 2 had a significantly worse overall survival [HR=1.24, 95%CI (1.04, 1.48), P=0.015]. ConclusionThe EZ-ALBI score is an easy and feasible classifying method to predict postoperative complications and survival of HCC.
Objective To observe the effect of forkhead box Q1(FOXQ1) short hairpin RNA (shRNA) on sensitivity of oxaliplatin chemotherapy in hepatpcellular carcinoma cell line SMMC-7721. Methods ① Complementary shRNA oligonucleotides targeting the FOXQ1 gene and negative control-shRNA were designed and inserted into lentiviral vector. shRNA lentivirus vectors were transfected into SMMC-7721 cells and the lentivirus vector with the best silencing effect was screened. ② SMMC-7721 cells were divided into interference group (SMMC-7721 cells were transfected with FOXQ1-shRNA-1), negative control group (SMMC-7721 cells were transfected with negative control-shRNA), and blank control group (SMMC-7721 cells did not received any treatment), and the expressions of FOXQ1 mRNA and its protein in SMMC-7721 cells were detected at 72 hours after transfection. ③ SMMC-7721 cells were divided into interference group, negative control group, blank control group, interference+oxaliplatin group, negative control+oxaliplatin group, and blank control+oxaliplatin group, apoptosis rates and viability of SMMC-7721 cells were detected at 48 hours after transfection. Results ① The expressions of FOXQ1 mRNA and its protein in SMMC-7721 cells of the FOXQ1-shRNA-1 group were both lower than those of the FOXQ1-shRNA-2 group and FOXQ1-shRNA-3 group (P<0.05), so the FOXQ1-shRNA-1 was the best lentiviral vector. ② Compared with the negative control group and the blank control group, the expressions of FOXQ1 mRNA and its protein of the interference group were both lower (P<0.05), but there was no significant difference between the negative control group and the blank control group (P>0.05). ③ Whether added oxaliplatin or not, compared with the negative control group and the blank control group, the apoptosis rates of the interference group were higher (P<0.05), but the viability of the interference group was lower (P<0.05), and there was no significant difference between the negative control group and the blank control group under the same condition (P>0.05). The apoptosis rates of groups (including the interference group, the negative control group, and the blank control group) which added oxaliplatin were higher than those groups didn’t add oxaliplatin (P<0.05), but viability of groups (including the interference group, the negative control group, and the blank control group) which added oxaliplatin was lower than those groups didn’t add oxaliplatin (P<0.05). Conclusion Down-regulation of expression of FOXQ1 by shRNA in hepatocellular carcinoma cell line SMMC-7721 can effectively induce apoptosis and increase sensitivity of SMMC-7721 cells to oxaliplatin.