【 Abstract 】 Objective To probe into the role of inositol 1, 4, 5-trisphosphate (IP3) and bax gene expression in apoptosis of HepG2 cells induced by genistein (Gen). Methods HepG2 cells were treated with different concentrations including 20, 40, 60 and 80 μ mol/L Gen as HepG2 cells cultured with 0 μmol/L Gen for 72 h was control; HepG2 cells were treated with 60 μmol/L Gen for 6, 12, 24, 48 and 72 h as HepG2 cells treated with 60 μmol/L Gen for 0 h was control. IP3 content, bax mRNA expression and apoptosis rate were assayed by IP3- ［ 3H ］ Birtrak assay, RT-PCR and flow cytometry, respectively. ResultsHepG2 cells incubated with each concentration of Gen for 72 h ， IP3 content was lower than that of control 〔 (17.7 ± 1.3), (11.2 ± 0.9), (4.9 ± 0.5), (4.8 ± 0.3) pmol/106 cells vs (29.4 ± 0.5) pmol/106 cells 〕 , P ＜ 0.01 ； bax mRNA expression (RI which was the gray degree multiply area of bax/the gray degree multiply area of β -actin) was higher than that of control (0.26 ± 0.02, 0.33 ± 0.05, 0.35 ± 0.06, 0.38 ± 0.05 vs 0.09 ± 0.01), P ＜ 0.01 ； The apoptosis rate was higher than that of control 〔 (10.1 ± 0.9)%, (18.7 ± 1.6)%, (28.7 ± 2.5)%, (27.9 ± 2.0)% vs (2.6 ± 0.1)% 〕 , P ＜ 0.01. HepG2 cells were incubated with 60 μ mol/L Gen for 6, 12, 24, 48 and 72 h ， IP3 content was lower than that of control 〔 (22.6 ± 0.9), (12.0 ± 1.4), (7.5 ± 0.8), (5.6 ± 0.5), (4.3 ± 0.6) pmol/106 cells vs (29.2 ± 0.6) pmol/106 cells 〕 , P ＜ 0.01 ； bax mRNA expression was higher than that of control incubated with 60 μ mol/L Gen for above 12 h (0.25 ± 0.06, 0.29 ± 0.02, 0.30 ± 0.02, 0.35 ± 0.04 vs 0.09 ± 0.01), P ＜ 0.01 ； The apoptosis rate in groups incubated with 60 μ mol/L Gen for 24, 48 and 72 h was significantly higher than that in control 〔 (7.4 ± 0.5)%, (20.5 ± 2.0)%, (30.7 ± 1.6)% vs (2.6 ± 0.1)% 〕 , P ＜ 0.01. ConclusionGen induces apoptosis of HepG2 cells by reducing IP3 production and increasing bax gene expression.
ObjectiveTo investigate the expression and clinical significance of cytochromes b561 (CYB561) in hepatocellular carcinoma (HCC). MethodsThe expression of CYB561 mRNA in HCC tissues and its relationship with prognosis were analyzed by database data. Immunohistochemistry (IHC) was used to detect the expression of CYB561 protein in 61 matched HCC tissues and their adjacent tissues, and the relationship between CYB561 protein expression and clinicopathological features and prognosis of HCC was analyzed. Kaplan-Meier method was used to draw the survival curve and Cox proportional hazard regression model was used to analyze the correlation between the expression of CYB561 protein and the prognosis of HCC. ResultsThe analysis of database data showed that the relative expression of CYB561 mRNA in HCC tissues was higher than that in adjacent tissues (P<0.001). Compared with HCC patients with negative expression of CYB561 mRNA, HCC patients with positive expression of CYB561 mRNA had worse overall survival (OS), relapse-free survival, progression-free survival and disease-free survival (all P<0.05). The results of IHC showed that the positive rates of CYB561 protein in HCC tissues and adjacent tissues were 57.38% (35/61) and 21.31%(13/61), respectively. The former was higher than the latter, with statistical significance (χ2=16.624, P<0.001). Survival analysis showed that the OS of patients with positive expression of CYB561 protein was worse than that of patients with negative expression (P<0.05). Multivariate Cox proportional hazard regression analysis showed that the positive expression of CYB561 protein was a risk factor for postoperative OS in HCC patients [HR=3.308, 95%CI (1.344, 8.144), P=0.009]. ConclusionCYB561 is positively expressed in HCC and suggests a worse survival, and may serve as a potential prognostic biomarker for HCC.
ObjectiveTo exclusively compare the short-and long-term outcomes of hepatic resection (HR) patients with multifocal tumors meeting the Milan criteria between locating in same and different sections. MethodsA total of 219 consecutive HR patients with multifocal tumors meeting the Milan criteria were divided into group SS (n=97, same section) and group DS (n=122, different sections) according to their anatomical location (Couinaud's segmentation). ResultsThe 1-, 3-, and 5-year overall survival (OS) and recurrence-free survival (RFS) rates were significantly higher in the group SS than those in the group DS (P < 0.05). The subgroup analysis showed that patients with 2 tumors and those undergoing en bloc resection were associated with better OS and RFS (P < 0.05). ConclusionsFor HCC patients with multifocal tumors meeting the Milan criteria, those with tumors locating in same hepatic section may have better longterm survival and lower HCC recurrence rates than those locating in different sections after HR.
Objective To dynamically study the formation of multidrug resistance(MDR) of human hepatocellular carcinoma cell SMMC-7721 induced by Adriamycin (ADM) and the role of multidrug resistance-associated protein(MRP) in its mechanisms.Methods Hepatocellular carcinoma cell SMMC-7721 was cultured in RPMI-1640 medium containing ADM with progressively increased concentration or directly cultured in medium containing different concentrations of ADM. Resistant index of drug-resistant variants of SMMC-7721 cell was determined by drawing cell dosage-reaction curves.Levels of MRP mRNA expression were detected by reverse transcription-polymerase chain reaction(RTPCR). Intracellular rubidomycin(DNR) concentration was examined by flow cytometry(FCM).Results With progressive increasing of ADM concentration in medium resistant index and levels of MRP mRNA expression were correspondingly increased but intracellular DNR concentration was markly reduced. When parental cells were directly cultured in medium containing different concentrations of ADM, the higher the ADM concentration, the higher the level of MRP mRNA expression, but intracellular DNR concentration was kept at the similar high level and most cells died. Conclusion ADM may progressively induce SMMC-7721 cell resistant to multiple chemotherapeutic drugs with reduced intracellular DNR accumulation associated with the overexpression of MRP gene.
Objective To explore the effects of overexpression of human tissue inhibitors of metalloproteinase-1 (hTIMP-1) on proliferation of human liver cancer cell line HepG2 in vitro. Methods A recombinant adenoviral vector containing full-length cDNA of hTIMP-1 was generated and transfected into HepG2. The viral titer was checked by measuring GFP, and the expression of hTIMP-1 in vitro was detected by the techniques of Western blot and semi-quantitative RT-PCR. The ultrastructure was observed by transmission electron microscope and the effects of overexpression of hTIMP-1 on proliferation of HepG2 in vitro was analyzed by MTT assay and growth curve. Results The resultant AdhTIMP-1 was successfully constructed and the expression of hTIMP-1 was detected by Western blot and RT-PCR. The growth and proliferation of HepG2, which had been transfected with AdhTIMP-1, was significantly inhibited. Conclusion The proliferation of HepG2 was markedly inhibited by recombinant adenovirus-mediated overexpression of hTIMP-1, which may pave the way for further application in liver gene therapy.
Laparoscopic hepatectomy is routinely used in the surgical treatment of hepatocellular carcinoma, and has formed a standardized operating procedure. Tumors located in the segments Ⅶ and Ⅷ of liver as well as the paracaval subsegment of caudate lobe are considered to be difficult sites for laparoscopic hepatectomy due to the deep anatomical location, proximity to important vascular structures, difficulty in exposing the visual field under laparoscopy, and limited operating space. Based on the experience of our team and related research reports, the authors analyzed and summarized countermeasures for the difficulties of laparoscopic hepatectomy in the treatment of hepatocellular carcinoma in difficult sites. Adhering to the tumor-centered and margin-based principles, accurate preoperative assessment, selection of the correct surgical approach, designing liver resection plane guided by hepatic vena while taking into account portal vein territory, and giving preference to ananatomical hepatectomy while preserving functional liver parenchyma as much as possible are the prerequisites for ensuring minimally invasive and oncology benefits for patients with hepatocellular carcinoma in difficult sites.
Objective To investigate the effects of adenovirus-mediated melanoma differentiation-associated gene-7 (mda-7)/IL-24 and/or adriamycin (ADM) on transplanted human hepatoma in nude mice and to explore a new way for hepatoma gene therapy combined with chemotherapy. Methods The recombinant adenovirus vector carrying Ad.mda-7 was constructed; Ad.mda-7 and/or ADM were injected into the tumor-bearing mice. Their effects on the growth of the tumor and the survival time of the mice were observed. The expressions of VEGF and TGF-β1 were detected by an immunohistochemistry method. Results Ad.mda-7 was constructed and expressed in vivo successfully. Compared with other three groups 〔control group (43.4±1.67) d, ADM group (64.2±4.14) d, Ad.mda-7 group (61.4±1.67) d〕, the mice treated with Ad.mda-7 combined with ADM had longer average survival time 〔(83.8±4.82) d, P＜0.01〕; the average size of tumor treated with Ad.mda-7 combined with ADM diminished significantly compared with that treated with ADM or Ad.mda-7 separately (P＜0.01). VEGF and TGF-β1 expressions of Ad.mda-7 group were (56.2±7.7)%, (35.2±4.5)%, respectively, and were lower than those in ADM group (VEGF: P＜0.05; TGF-β1: P＜0.01). VEGF expression of Ad.mda-7+ADM group was (37.3±5.0)%, and was significantly lower than that in other three groups (P＜0.01). TGF-β1 expression of Ad.mda-7+ADM group was (31.2±3.1)% and significantly lower than that in control group and ADM group (P＜0.01), however, there was no significant difference compared with Ad.mda-7 group (Pgt;0.05). Conclusion Ad.mda-7 combined with ADM has b antitumor potency and synergistic effects and suppresses the growth of human HCC xenograft in nude mice, possibly by inducing the apoptosis of hepatoma cell lines and suppressing tumor angiogenesis.
ObjectiveTo summarize the current status and progress of surgical treatment for postoperative recurrent hepatocellular carcinoma (HCC).MethodThe literatures about studies of surgical treatment of postoperative recurrent HCC were reviewed.ResultsThe surgical operation was an effective method for the treatment of recurrent HCC. The operation methods included re-hepatectomy and salvage liver transplantation. There was no uniform standard for the indication of re-hepatectomy, but the basic principles were the same. At present, the indication of salvage liver transplantation was mainly based on Milan criteria. For patients with recurrent HCC who met the operation indications, surgical operation could improve the long-term survival rate of patients and benefit the patients.ConclusionIt migh prolong the survival time and improve the long-term survival rate of patients with recurrent HCC when the appropriate patients and reasonable surgical methods are chosen according to the surgical indications, the tumor situation of initial hepatectomy, postoperative recurrence time, and other factors.
Two hundred and thirty patients with solid hepatic space-occupying lesions (SHSOL), on whom hepatic resection was performed in Zhongshan hospital, were analyzed. We found that liver cirrhosis could be a diagnostic marker of hepatocellular carcinoma in patients with SHSOL, for which the sensitivity being 85.2%, the specificity 96.3%, and the positive predictive value 98.7%.
ObjectiveTo summarize the experience of the whole process management of hepatocellular carcinoma (HCC) patients with high-risk of recurrence and metastasis based on the multidisciplinary team (MDT) mode, and to improve the clinicians’ understanding of the concept of whole process management, so as to improve the survival rate of patients with HCC. MethodThe clinicopathologic data of a HCC patient with high-risk of recurrence and metastasis admitted to the Division of Liver Surgery, Department of General Surgery, West China Hospital of Sichuan University were retrospectively analyzed. ResultsA 52-year-old male patient was diagnosed with HCC with intrahepatic metastasis (China liver cancer staging Ⅱ b, Barcelona Clinic Liver Cancer stage B) after admission due to “epigastric discomfort for 1+-month and liver occupying for 1+-week”. Through discussion by the MDT mode, the allogeneic liver transplantation was performed after successful downstaging following two conversion therapies. No serious complications occurred after operation, and the patient was discharged on the 23rd day after operation. Up to now, pulmonary bacterial and fungal infections and pulmonary metastases had been found during the postoperative follow-up. After anti-infective therapy and targeted therapy combined with radiotherapy, the patient was significantly relieved, had survived for 34 months after operation, and was still under regular follow-up. ConclusionsFor HCC patients with high-risk of recurrence and metastasis, MDT mode has a good clinical benefit for the whole process management of patient. Through the MDT model, the diagnosis, treatment, and follow-up of HCC are organically integrated, and the patient’ s diagnosis and treatment plans are dynamically adjusted to realize the whole process management of HCC patient, and to raise the survival rate and improve quality of life of HCC patient.