ABSTRACT
Bariatric surgery is effective in treating different components of metabolic syndrome including obesity, type 2 diabetes mellitus (T2DM), and hyperlipidemia. But there is no consensus on the ideal biliopancreatic and Roux limb length. This study aimed to explore the effect of biliopancreatic limb and Roux limb lengths during laparoscopic Roux-en-Y gastric bypass (LRYGB) procedures on weight loss and T2DM control.We studied the clinical records of 58 patients with metabolic syndrome, T2DM, and body mass index (BMI) 32 to 50âkg/m who underwent LRYGB in our hospital. The short limb group (Group A) underwent LRYGB with a limb length of 160 to 200âcm (nâ=â31) and the long limb group (Group B) underwent LRYGB with a limb length of 210 to 240âcm (nâ=â27) were compared.The occurrence of acute or chronic internal hernia in Group B was higher than that in Group A (Pâ=â.026). Twelve months after surgery, patients from the 2 groups were also observed with reduction in BMI, percent excess weight loss (EWL), preoperative FPG, and HbA1c as compared with these indicators before surgery. However, the differences of these indicators between 2 groups were not significant at the time point of before and 3, 6, 12 months after surgery.LRYGB had significant effects on weight loss and diabetes control in obese T2DM patients. However, there was no significant difference in the short term on weight loss and diabetes control in the patients receiving different limb lengths.
Subject(s)
Anastomosis, Roux-en-Y/methods , Biliopancreatic Diversion/methods , Diabetes Mellitus, Type 2/surgery , Gastric Bypass/methods , Obesity/surgery , Adult , Body Mass Index , China , Diabetes Mellitus, Type 2/complications , Female , Humans , Laparoscopy/methods , Male , Middle Aged , Obesity/complications , Postoperative Period , Time Factors , Treatment Outcome , Weight Loss , Young AdultABSTRACT
Recent studies have indicated that telomerase activity promotes cancer invasion and metastasis, but the underlying mechanism remains unclear. Several studies have shown that expression of exogenous human telomerase reverse transcriptase (hTERT) can promote motility and invasiveness among telomerase-negative tumor cells, and inhibition of endogenous telomerase activity can reduce invasiveness in tumor cells. However, whether overexpression of hTERT can further enhance the motility and invasiveness of telomerasepositive tumor cells has yet to be determined. In the present study, we showed that stable overexpression of hTERT can increase telomerase activity and telomere length, which significantly promotes the invasive and metastatic potential of telomerasepositive HepG2 cells but does not affect cell proliferation. Further analysis suggested that enhanced invasiveness and metastasis may act through corresponding upregulation of mRNA and protein expression of matrix metalloproteinase 9 (MMP9) and Ras homolog gene family member C (RhoC). Our study indicated that exogenous expression of hTERT may promote invasiveness and metastasis through upregulation of MMP9 and RhoC.
Subject(s)
Cell Movement , Cell Proliferation , Gene Expression Regulation, Neoplastic , Telomerase/metabolism , Blotting, Western , Genetic Vectors , Hep G2 Cells , Humans , In Vitro Techniques , Matrix Metalloproteinase 9/genetics , Matrix Metalloproteinase 9/metabolism , Neoplasm Invasiveness , RNA, Messenger/genetics , Real-Time Polymerase Chain Reaction , Retroviridae/genetics , Reverse Transcriptase Polymerase Chain Reaction , Telomerase/genetics , rho GTP-Binding Proteins/genetics , rho GTP-Binding Proteins/metabolism , rhoC GTP-Binding ProteinABSTRACT
BACKGROUND/AIMS: Hepatocellular carcinoma (HCC) is a prevalent malignant tumor. Tumor markers are very useful in early diagnosis; however a single marker is rather limited. We launched a test to increase the diagnostic sensitivity through the combined detection. METHODOLOGY: Serum concentration of three tumor-markers, Glypican-3 (GPC-3), Human-Cervical-Cancer-Oncogene (HCCR) and a-fetoprotein (AFP), were determined in 189 samples: 101 cases of HCC, 40 cases of cirrhosis, 18 cases of hepatitis and 30 cases of control healthy donors. Every marker was evaluated for its diagnostic value by one-way-analysis-of-variance and receiver-operating-characteristics analysis. RESULTS: GPC-3 was the best marker with an area under the curve (AUC) of 0.892; using 26.8ng/mL as the cut-off for HCC diagnosis, GPC-3 has a sensitivity of 51.5% and maintains a specificity of 92.8%. HCCR, with an AUC of 0.831, can reach a sensitivity of 22.8% and maintain a specificity of 90.9% if the cut-off is set as 58.8mAU/mL. With an AUC of 0.827, the efficacy and sensitivity of AFP were 36.6% and 98.5% when using 199.3ng/mL as the cut-off. No significant correlation was found between these three markers. Simultaneously detecting three markers can significantly increases the sensitivity to 80.2%, much higher than AFP alone. CONCLUSIONS: GPC-3 and HCCR are useful tumor markers complementary to AFP for clinical diagnosis of HCC.
Subject(s)
Carcinoma, Hepatocellular/blood , Glypicans/blood , Liver Neoplasms/blood , Proto-Oncogene Proteins/blood , alpha-Fetoproteins/metabolism , Adult , Analysis of Variance , Area Under Curve , Biomarkers, Tumor/blood , Carcinoma, Hepatocellular/diagnosis , Enzyme-Linked Immunosorbent Assay , Female , Humans , Liver Neoplasms/diagnosis , Male , Middle Aged , Predictive Value of Tests , ROC Curve , Sensitivity and SpecificityABSTRACT
Therapeutic numbers of antigen-specific cytotoxic T lymphocytes (CTLs) are key effectors in successful adoptive immunotherapy. However, efficient and reproducible methods to meet the qualification remain poor. To address this issue, we designed the artificial antigen-presenting cell (aAPC) system based on poly(lactic-co-glycolic acid) (PLGA). A modified emulsion method was used for the preparation of PLGA particles encapsulating interleukin-2 (IL-2). Biotinylated molecular ligands for recognition and co-stimulation of T cells were attached to the particle surface through the binding of avidin-biotin. These formed the aAPC system. The function of aAPCs in the proliferation of specific CTLs against human Flu antigen was detected by enzyme-linked immunospot assay (ELISPOT) and MTT staining methods. Finally, we successfully prepared this suitable aAPC system. The results show that IL-2 is released from aAPCs in a sustained manner over 30 days. This dramatically improves the stimulatory capacity of this system as compared to the effect of exogenous addition of cytokine. In addition, our aAPCs promote the proliferation of Flu antigen-specific CTLs more effectively than the autologous cellular APCs. Here, this aAPC platform is proved to be suitable for expansion of human antigen-specific T cells.
Subject(s)
Antigen-Presenting Cells/immunology , Artificial Cells/immunology , Immunotherapy, Adoptive , Peptide Fragments/immunology , T-Lymphocytes, Cytotoxic/immunology , Viral Matrix Proteins/immunology , Avidin/chemistry , Biotin/chemistry , Biotinylation , Capsules , Enzyme-Linked Immunospot Assay , Humans , Interleukin-2/chemistry , Interleukin-2/immunology , Lactic Acid/chemistry , Ligands , Lymphocyte Activation , Microspheres , Polyglycolic Acid/chemistry , Polylactic Acid-Polyglycolic Acid Copolymer , T-Lymphocytes, Cytotoxic/transplantationABSTRACT
AIM: To evaluate the diagnostic value of cancer-testis antigen (CTA) mRNA in peripheral blood samples from hepatocellular carcinoma (HCC) patients. METHODS: Peripheral blood samples were taken from 90 patients with HCC before operation. Expression of melanoma antigen-1 (MAGE-1), synovial sarcoma X breakpoint-1 (SSX-1), and cancer-testis-associated protein of 11 kDa (CTp11) mRNA in peripheral blood mononuclear cells (PBMC) was tested by nested reverse transcripts-polymerase chain reaction (RT-PCR). Serum alpha-fetoprotein (AFP) in these patients was also determined. RESULTS: The positive rate of MAGE-1, SSX-1 and CTp11 transcripts was 37.7%, 34.4%, 31.1% in PBMC samples, and 74.4%, 73.3%, 62.2% in their resected tumor samples, respectively. The positive rate for at least one of the transcripts of three CTA genes was 66.7% in PBMC samples and 91.1% in their resected tumor samples. MAGE-1, SSX-1 and/or CTp11 mRNA were not detected in the PBMC of those patients from whom the resected tumor samples were MAGE-1, SSX-1 and/or CTp11 mRNA negative, nor in the PBMC samples from 20 healthy donors and 10 cirrhotic patients. Among the 90 patients, the serum AFP in 44 patients met the general diagnostic standard (AFP > 400 microg/L) for HCC, and was negative (AFP < or = 20 microg/L) or positive with a low concentration (20 microg/L < AFP < or = 400 microg/L) in the other patients. The positive rate for at least one of the transcripts of three CTA genes in PBMC samples from the AFP negative or positive patients with a low concentration was 69.2% and 45.0%, respectively. Of the 90 patients, 71 (78.9%) were diagnosed as HCC by nested RT-PCR and serum AFP. Although the positive rate for at least one of the transcripts of three CTA genes in PBMC samples from 53 patients at TNM stage III or IV was obviously higher than that in PBMC samples from 37 patients at stage I or II (77.9% vs 51.4%, P = 0.010), the CTA mRNA was detected in 41.7% and 56.0% of PBMC samples from HCC patients at stages I and II, respectively. CONCLUSION: Detecting MAGE-1, SSX-1 and CTp11 mRNA in PBMC improves the total diagnostic rate of HCC.
Subject(s)
Antigens, Neoplasm , Carcinoma, Hepatocellular , Liver Neoplasms , Neoplasm Proteins , Adolescent , Adult , Aged , Antigens, Neoplasm/blood , Antigens, Neoplasm/genetics , Carcinoma, Hepatocellular/blood , Carcinoma, Hepatocellular/diagnosis , Carcinoma, Hepatocellular/genetics , Cell Line, Tumor , Female , Humans , Liver Neoplasms/blood , Liver Neoplasms/diagnosis , Liver Neoplasms/genetics , Male , Middle Aged , Neoplasm Proteins/blood , Neoplasm Proteins/genetics , Reverse Transcriptase Polymerase Chain Reaction/methods , Young AdultABSTRACT
To study the effects of Smad4 on liver fibrosis and hepatocarcinogenesis in mice treated with CCl(4)/ethanol. The wild-type mice (Smad4 +/+) and the Smad4 knockout mice (Smad4 +/-) were injected subcutaneously with carbon tetrachloride(CCl(4))/ethanol twice a week for twenty weeks. The expression of Smad4, TGFbeta1, Smad2, Smad3, Smad6, TIMP1, MMP2 and MMP9 was detected by RT-PCR. In the cirrhotic liver, the expression of Smad4 mRNA was significantly higher than that in the normal liver. Comparing with wild-type mice (Smad4 +/+), the TGFbeta1-Smad4 signaling was markedly attenuated in the Smad4 knockout mice (Smad4 +/-). After induction by CCl(4)/ethanol, the hepatic fibrosis in the Smad4 knockout mice (Smad4 +/-) was obviously alleviated compared with the wild-type mice (Smad4 +/+), and the incidence rate of hepatocarcinogenesis of the former was also lower than that of the latter(32.0% vs 41.9%). These results indicate that knocking out Smad4 can delay the progression of liver fibrosis and liver cancer.
Subject(s)
Liver Cirrhosis, Experimental/pathology , Liver Neoplasms, Experimental/pathology , Signal Transduction , Smad4 Protein/genetics , Transforming Growth Factor beta1/metabolism , Animals , Carbon Tetrachloride/administration & dosage , Disease Models, Animal , Ethanol/administration & dosage , Female , Liver Cirrhosis, Experimental/chemically induced , Liver Cirrhosis, Experimental/metabolism , Liver Neoplasms, Experimental/chemically induced , Liver Neoplasms, Experimental/metabolism , Male , Mice , Mice, Knockout , RNA, Messenger/genetics , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Smad Proteins/genetics , Smad Proteins/metabolism , Smad4 Protein/metabolism , Tissue Inhibitor of Metalloproteinase-1/genetics , Tissue Inhibitor of Metalloproteinase-1/metabolism , Transforming Growth Factor beta1/geneticsABSTRACT
OBJECTIVE: To observe the effects of galectin-3 on proliferation and apoptosis of hepatic stellate cells. METHODS: RT-PCR and Western blot were used to detect the expression of galectin-3 in hepatic stellate cells. Short hairpin DNA targeting galectin-3 of rat was was ligated into the recombinant vector pGCsilencer U6/Neo/GFP/shRNA plasmid. Then the plasmid was transfected into rat hepatic stellate cells. RT-PCR and Western blot were used to detect the interfering efficiency. Cell proliferation level was observed by CCK8 method at 24, 48 and 72 hours after transfection. Cell apoptosis was measured by Annexin V/PI-labeled flow cytometric analysis. RESULTS: Expression of galectin-3 in HSC was verified by both RT-PCR and Western blot. The recombinant vector was successfully constructed and verified, and was transfected into rat hepatic stellate cells. Western Blot and RT-PCR results demonstrated that the expression level of Galectin-3 was significantly down-regulated in galectin-3 shRNA transfected cells compared to control vector transferred cells. CCK8 assay indicated that proliferation of Galectin-3 knockdown cells was lower than that of control cells 48 and 72 hours post-transfection. Apoptotic cells in shRNA-interfering group were higher than those in control group both in early stage and advanced stage. CONCLUSION: Hepatic stellate cells can express galectin-3. Inhibition of galectin-3 using RNAi technique can suppress proliferation and induce apoptosis in HSC.
Subject(s)
Apoptosis , Cell Proliferation , Galectin 3/metabolism , Hepatic Stellate Cells/metabolism , RNA, Small Interfering/genetics , Animals , Cell Line , Down-Regulation , Flow Cytometry , Galectin 3/genetics , Genetic Vectors , Hepatic Stellate Cells/cytology , Liver Cirrhosis/pathology , Plasmids/genetics , RNA Interference , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rats , Reverse Transcriptase Polymerase Chain Reaction , TransfectionABSTRACT
The prognosis of hepatocellular carcinoma (HCC) after surgery is poor due to its high recurrence rate. In order to unfold the mechanism of different recurrent-free survival (RFS) times following resection, expression profiling of tumor tissues from 32 HCC patients with different RFS time were used to identify differential expression of individual genes and signaling pathway components correlated with RFS time. Quantitative RT-PCR, Western blotting, and immunohistochemistry were used to validate the expression of selected genes. Up-regulation of several immune related genes and pathways, especially HLA II-related antigen presenting pathways, significantly correlated with longer RFS time. The expression of MHCII molecules were found to be mainly located in either CD68+ cells or CD45+ cells, and their expression significantly correlated with the expression of CIITA (HLA II genes transactivator) in the tumor. The results suggest that the high expression level of CIITA and MHCII molecules in hepatocellular carcinoma tissue is an effective prognostic marker for longer RFS time in HCC.
Subject(s)
Carcinoma, Hepatocellular/genetics , Liver Neoplasms/genetics , Nuclear Proteins/biosynthesis , Trans-Activators/biosynthesis , Biomarkers, Tumor/biosynthesis , Biomarkers, Tumor/genetics , Biomarkers, Tumor/immunology , Carcinoma, Hepatocellular/immunology , Carcinoma, Hepatocellular/metabolism , Cluster Analysis , Disease-Free Survival , Female , Gene Expression , Genes, MHC Class II , Humans , Immunohistochemistry , Liver Neoplasms/immunology , Liver Neoplasms/metabolism , Male , Microarray Analysis , Middle Aged , Nuclear Proteins/genetics , Reverse Transcriptase Polymerase Chain Reaction , Survival Analysis , Trans-Activators/geneticsABSTRACT
Increasing evidence indicates the immunosuppressive nature of the local environment in tumor. The present study was focused on analyzing the immune status within hepatocellular carcinoma. In contrast to the increasing number of CD4(+) T cells, CD8(+), CD3(-)CD56(+), CD3(+)CD56(+), and gammadeltaT cells were all found to be under-represented in tumor infiltrating lymphocytes. Notably, the relative abundance of CD3(+)CD56(+) cells appeared to be correlated with patient survival. Functional analysis demonstrated that CD4(+) cells in the tumor tended to produce more IL-10 but less IFN-gamma, whereas CD8(+) cells showed impaired capacity for the production of both IFN-gamma and perforin. Consistent with previous reports, we observed a significant increase of Foxp3(+) cells in the tumor tissue. Intriguingly, although over 90% of CD4(+)CD25(high) cells were found to be Foxp3(+), the majority of Foxp3(+) cells were identified in the CD4(+)CD25(medium) and CD4(+)CD25(-) subsets. In support of its role as a negative regulator, CD4(+)CD25(high) cells suppressed the proliferation of CD4(+)CD25(-) cells isolated from the same tissues in an APC dependent manner. In conclusion, the tumor microenvironment of hepatocellular carcinoma is featured by the presence of multiple immunosuppressive factors.
Subject(s)
Carcinoma, Hepatocellular/immunology , Immunosuppressive Agents/metabolism , Lymphocytes, Tumor-Infiltrating/immunology , Adult , Aged , Aged, 80 and over , CD3 Complex/immunology , CD4 Antigens/immunology , Carcinoma, Hepatocellular/pathology , Cell Proliferation , Female , Flow Cytometry , Forkhead Transcription Factors/metabolism , Humans , Immunosuppressive Agents/immunology , Interferon-gamma/metabolism , Interleukin-10/immunology , Lymphocyte Activation , Male , Middle Aged , T-Lymphocytes/immunology , T-Lymphocytes/pathology , T-Lymphocytes, Regulatory/immunologyABSTRACT
NY-ESO-1 is an important member of cancer-testis antigen family and is widely distributed among many cancer types. As a tumor-specific antigen with the strongest immunogenicity so far identified, it can induce spontaneous antibody and T-cell responses in patients with NY-ESO-1-positive tumors. Therefore, it has been a good vaccine candidate in the immunotherapy against many malignancies. This article reviews the recent research advances in NY-ESO-1 and its relevant vaccines.
Subject(s)
Antigens, Neoplasm/therapeutic use , Immunotherapy , Membrane Proteins/therapeutic use , Neoplasms/immunology , Neoplasms/therapy , Antigens, Neoplasm/genetics , Antigens, Neoplasm/immunology , Cancer Vaccines/immunology , Cancer Vaccines/therapeutic use , Clinical Trials as Topic , Humans , Membrane Proteins/genetics , Membrane Proteins/immunology , Neoplasms/geneticsABSTRACT
The cancer testis (CT) antigen HCA587 is highly expressed in human hepatocellular carcinoma (HCC) and induces specific T-cell responses in a significant proportion of HCC patients. To explore its potential in cancer immunotherapy, a reverse immunology approach was adopted to identify HCA587-derived HLA-A( *)0201-restricted epitopes. Multiple peptides with a top ranking in various prediction programs were thus synthesized and three of them-p248-256, p140-149 and p144-152-were found to bind to HLA-A(*)0201 molecules with a high affinity and effectively induced a recall response of CD8+ T cells, which were either primed in vitro with the HCA587 antigen or directly isolated from HCC patients bearing HCA587+ tumors. Notably, these peptide-specific CD8+ T cells exhibited potent cytotoxic activity over HCA587+ tumor cells. Taken together, the present study has identified three new HLA-A(*)0201-restricted cytotoxic T cell epitopes in the CT antigen HCA587, which may serve as targets for peptide-based immunotherapy for HCC patients.
Subject(s)
Antigens, Neoplasm/analysis , Carcinoma, Hepatocellular/immunology , Epitopes, T-Lymphocyte/analysis , HLA-A Antigens/analysis , Liver Neoplasms/immunology , Neoplasm Proteins/analysis , T-Lymphocytes, Cytotoxic/immunology , Amino Acid Sequence , Antigens, Neoplasm/immunology , Carcinoma, Hepatocellular/therapy , Cell Line, Tumor , Epitope Mapping , Epitopes, T-Lymphocyte/immunology , HLA-A Antigens/immunology , HLA-A2 Antigen , Humans , Immunotherapy , Liver Neoplasms/therapy , Male , Molecular Sequence Data , Neoplasm Proteins/immunology , Peptides/chemical synthesis , Peptides/immunologyABSTRACT
Immunoselection and tumor evasion constitutes one of the major obstacles in cancer immunotherapy. A potential solution to this problem is the development of polyvalent vaccines, and the identification of more tumor-specific antigens is a prerequisite for the development of cancer vaccines. To identify novel tumor-specific antigens, suppression subtractive hybridization (SSH) was performed to isolate genes differentially expressed in human hepatocellular cancer (HCC) tissues. PLAC1 (PLACenta-specific 1) was one of the genes identified highly expressed in HCC tissues but not in paired noncancerous tissues. Further analyses revealed its expression in several other types of cancer tissues as well as tumor cell lines, but not in normal tissues except for placenta. Among HCC samples tested, 32% (22/69) showed PLAC1 mRNA expression while the protein was detected in 23.3% (7/30). A serological survey revealed that 3.8% (4/101) of HCC patients had anti-PLAC1 antibody response, suggesting the immunogenicity of PLAC1 in HCC patients. PLAC1 represents a new class of tumor associated antigen with restricted expression in placenta and cancer tissues, that may serve as a target for cancer vaccination.
Subject(s)
Antibodies, Neoplasm/blood , Carcinoma, Hepatocellular/immunology , Liver Neoplasms/immunology , Pregnancy Proteins/immunology , Antibody Formation , Blotting, Northern , Blotting, Western , Cancer Vaccines/immunology , Gene Expression Regulation, Neoplastic , Humans , Immunohistochemistry , Nucleic Acid Hybridization/methods , Recombinant Proteins/immunology , Reverse Transcriptase Polymerase Chain Reaction , Subtraction TechniqueABSTRACT
OBJECTIVE: To identify a naturally presented HLA-A2-restricted epitope of MAGE-A3 antigen, FLWGPRALV (MAGE-A3(271 - 279)), on the surface of a human hepatocellular carcinoma (HCC) cell line HLE. METHODS: Synthetic peptide FLWGPRALV, served as positive control target, was analyzed by HPLC and HPLC-ESI-TOF-MSMS, in order to determine its HPLC elution time, mass-spectrometric characteristics and the lowest detection limitation by the two approaches. 3 x 10(9) HLE cells were collected, peptides naturally presented by major histocompatibility complex (MHC) molecules on the cell surface were isolated by mild acid elution, and concentrated by lyophilization, then the mixtures of peptides were fractioned by HPLC. The ingredient ranged from 2 min before the elution time determined by the synthetic peptide to 2 min after that was collected, concentrated by lyophilization, and analyzed by HPLC-ESI-TOF-MSMS, to identify the existence of the MAGE-A3(271 - 279) peptide. RESULTS: The HPLC-ESI-TOF-MSMS detection provided an evidence for the existence of a doubly charged ion of (m/z)(2) 529.9, which was further analyzed by collision induced dissociation. The doubly charged ion was ultimately identified as the MAGE-A3(271 - 279) peptide, its amino sequence was FLWGPRALV and its molecular weight was 1058.4 Da. CONCLUSIONS: MAGE-A3(271 - 279) epitope could be naturally presented by HLA-A2 molecules to the surface of HCC cell line and MAGE-A3(271 - 279) peptide may have potential immunotherapeutic value in HCC patients.
Subject(s)
Antigens, Neoplasm/analysis , Epitopes, T-Lymphocyte/analysis , Neoplasm Proteins/analysis , Amino Acid Sequence , Antigen Presentation , Antigens, Neoplasm/isolation & purification , Carcinoma, Hepatocellular/immunology , Carcinoma, Hepatocellular/pathology , Cell Line, Tumor , Chromatography, High Pressure Liquid , Epitopes, T-Lymphocyte/isolation & purification , HLA-A2 Antigen/immunology , Humans , Liver Neoplasms/immunology , Liver Neoplasms/pathology , Mass Spectrometry , Neoplasm Proteins/isolation & purificationABSTRACT
The MAGE-A3 protein, one of the promising tumor antigens for immunotherapy, is highly expressed in human hepatocellular carcinoma (HCC). In this study, we estimated the specific CD8(+) T cell immune response to MAGE-A3 p271-279 peptide (M3(271)) in the peripheral blood of HCC patients without antigen vaccination in order to evaluate its immunotherapeutic potential in these patients. After expansion in vitro, the functional IFN-gamma producing M3(271) specific CD8(+) T cells were detected in 30.8% (8/26) of HLA-A2(+)MAGE-A3(+) HCC patients. The effector CD8(+ )T cells could release cytotoxic molecules of granzyme B and perforin after restimulation with natural HLA-A2(+)MAGE-A3(+) HCC cell lines in the samples tested. The functional supertype of HLA-A2 in the presentation of HLA-A*0201 restricted M3(271) peptide has been identified in the Chinese HCC patients of Han ethnicity, that widely expanded the applicability of this tumor peptide vaccine in Chinese HCC patients. Thus, the functionally detectable pre-existence of M3(271)-specific CD8(+) T cells in HCC patients makes M3(271) a potential target for immunotherapy in these patients. The responsive CD8(+ )T cells to both NY-ESO-1 and MAGE-A3 antigens provide a rationale for the application of a bivalent vaccine in HCC patients with tumors expressing both antigens.
Subject(s)
Antigens, Neoplasm/chemistry , CD8-Positive T-Lymphocytes/metabolism , Cancer Vaccines/chemistry , Carcinoma, Hepatocellular/metabolism , HLA-A2 Antigen/biosynthesis , Liver Neoplasms/metabolism , Neoplasm Proteins/chemistry , Antineoplastic Agents/chemistry , China , Epitopes/chemistry , HLA-A2 Antigen/chemistry , Humans , Immunotherapy/methods , Leukocyte Common Antigens/biosynthesis , Peptides/chemistry , T-Lymphocytes/metabolismABSTRACT
BACKGROUND: Hepatic stellate cell (HSC) plays a key role in hepatic fibrosis. This study was undertaken to investigate the expression of 5-hydroxytamine receptors in HSC and the effect of 5-hydroxytamine on biological characteristics of HSC. METHODS: Liver ex vivo perfusion of collagenase and density gradient centrifugation were used to isolate HSCs. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the expression of 5-hydroxytamine receptor subtypes 1A, 2A, 2B and 3. Western blot hybridization was used to elucidate the effect of 5-hydroxytamine and its 2A receptor antagonist ketanserin and 3 receptor antagonist ondanosetron on the expression of transforming growth factor-beta1 (TGF-beta1) and Smad4 in HSC. RESULTS: HSC expressed 5-hydroxytamine receptor subtypes 1A, 2A and 2B. 5-hydroxytamine significantly increased the expression of TGF-beta1 and Smad4 in HSC (P<0.05). This action can be antagonized by ketanserin, not by ondanosetron. CONCLUSIONS: HSC expresses 5-hydroxytamine receptors. 5-Hydroxytamine could effect the biological characteristics of HSC through its receptor mediation, and may play a role in the pathogenesis of liver cirrhosis and portal hypertension.
Subject(s)
Liver/drug effects , Serotonin Antagonists/pharmacology , Serotonin/pharmacology , Animals , Blotting, Western , Cells, Cultured , Gene Expression/drug effects , Ketanserin/pharmacology , Liver/cytology , Liver/metabolism , Liver Cirrhosis/etiology , Male , Ondansetron/pharmacology , RNA/genetics , Rats , Rats, Wistar , Receptors, Serotonin/biosynthesis , Receptors, Serotonin/drug effects , Receptors, Serotonin/genetics , Reverse Transcriptase Polymerase Chain Reaction , Smad4 Protein/biosynthesis , Smad4 Protein/drug effects , Transforming Growth Factor beta/biosynthesis , Transforming Growth Factor beta/drug effects , Transforming Growth Factor beta1ABSTRACT
OBJECTIVE: To construct dendritomas by fusion of human dendritic cells with HLE cells, a human hepatocellular carcinoma cell line. METHODS: HLE cells were cultured in RPMI 1640 with 15% FCS. Human dendritic cells (DCs) were obtained from peripheral blood monocytes cultured in the presence of GM-CSF and IL-4 for 7 days, matured with TNF-alpha and PGE(2) for 2 days. The DCs and HLE cells were labeled with green fluorescence dye PKH67-GL and red fluorescence dye PKH26-GL, respectively, and fused in 50% polyethylene glycol (PEG) + 10% dimethyl sulfoxide (DMSO) to generate dendritomas for rapid fluorescence-activated cell sorting (FACS). RESULTS: Dendritomas with dual red-green fluorescence were constructed successfully, and FACS analysis showed the effective fusion rate was 16.8%. CONCLUSION: With fluorescence dyes PKH67-GL and PKH26-GL as fusion markers, dendritomas for rapid fluorescence-activated cell sorting are constructed, which may throw new light on immunotherapy of hepatocellular carcinoma.
Subject(s)
Carcinoma, Hepatocellular/pathology , Cell Fusion/methods , Dendritic Cells/cytology , Liver Neoplasms/pathology , Cancer Vaccines/biosynthesis , Cell Line, Tumor , Cells, Cultured , Humans , Hybrid CellsABSTRACT
AIM: To study the effect of rosiglitazone, which is a ligand of peroxisome proliferator-activated receptor gamma (PPARgamma), on the expression of PPARgamma in hepatic stellate cells (HSCs) and on the biological characteristics of HSCs. METHODS: The activated HSCs were divided into three groups: control group, 3 micromol/L rosiglitazone group, and 10 micromol/L rosiglitazone group. The expression of PPARgamma, alpha-smooth muscle actin (alpha-SMA), and type I and III collagen was detected by RT-PCR, Western blot and immunocytochemical staining, respectively. Cell proliferation was determined with methylthiazolyltetrazolium (MTT) colorimetric assay. Cell apoptosis was demonstrated with flow cytometry. RESULTS: The expression of PPARgamma at mRNA and protein level markedly increased in HSCs of 10 micromol/L rosiglitazone group (t value was 10.870 and 4.627 respectively, P<0.01 in both). The proliferation of HSCs in 10 micromol/L rosiglitazone group decreased significantly (t = 5.542, P<0.01), alpha-SMA expression level and type I collagen synthesis ability were also reduced vs controls (t value = 10.256 and 14.627 respectively, P<0.01 in both). The apoptotic rate of HSCs significantly increased in 10 micromol/L rosiglitazone group vs control (chi(2) = 16.682, P<0.01). CONCLUSION: By increasing expression of PPARgamma in activated HSCs, rosiglitazone, an agonist of PPARgamma, decreases alpha-SMA expression and type I collagen synthesis, inhibits cell proliferation, and induces cell apoptosis.
Subject(s)
Hepatocytes/drug effects , PPAR gamma/agonists , Thiazolidinediones/pharmacology , Actins/metabolism , Animals , Apoptosis/drug effects , Base Sequence , Cell Proliferation/drug effects , Collagen/metabolism , DNA/genetics , Hepatocytes/cytology , Hepatocytes/metabolism , In Vitro Techniques , Ligands , PPAR gamma/genetics , PPAR gamma/metabolism , Rats , RosiglitazoneABSTRACT
AIM: To review the experience in surgery for 508 patients with portal hypertension and to explore the selection of reasonable operation under different conditions. METHODS: The data of 508 patients with portal hypertension treated surgically in 1991-2001 in our centers were analyzed. Of the 508 patients, 256 were treated with portaazygous devascularization (PAD), 167 with portasystemic shunt (PSS), 62 with selective shunt (SS), 11 with combined portasystemic shunt and portaazygous devascularization (PSS+PAD), 9 with liver transplantation (LT), 3 with union operation for hepatic carcinoma and portal hypertension (HCC+PH). RESULTS: In the 167 patients treated with PSS, free portal pressure (FPP) was significantly higher in the patients with a longer diameter of the anastomotic stoma than in those with a shorter diameter before the operation (P < 0.01). After the operation, FPP in the former patients markedly decreased compared to the latter ones (P < 0.01). The incidence rate of hemorrhage in patients treated with PAD, PSS, SS, PSS+PAD, and HCC+PH was 21.09% (54/256), 13.77 (23/167), 11.29 (7/62), 36.36% (4/11), and 100% (3/3), respectively. The incidence rate of hepatic encephalopathy was 3.91% (10/256), 9.58% (16/167), 4.84% (3/62), 9.09% (1/11), and 100% (3/3), respectively while the operative mortality was 5.49% (15/256), 4.22% (7/167), 4.84% (3/62), 9.09% (1/11), and 66.67% (2/3) respectively. The operative mortality of liver transplantation was 22.22% (2/9). CONCLUSION: Five kinds of operation in surgical treatment of portal hypertension have their advantages and disadvantages. Therefore, the selection of operation should be based on the actual needs of the patients.
Subject(s)
Hypertension, Portal/mortality , Hypertension, Portal/surgery , Portasystemic Shunt, Surgical/mortality , Adolescent , Adult , Aged , Child , Child, Preschool , Female , Hemorrhage/mortality , Hemorrhage/surgery , Humans , Liver Neoplasms/mortality , Liver Neoplasms/surgery , Liver Transplantation/mortality , Male , Middle Aged , Postoperative Complications/mortality , Retrospective StudiesABSTRACT
OBJECTIVES: To investigate the effect of interlukin-10 (IL-10) on expression and secretion of collagen I, IV in rat's hepatic stellate cells (HSC) of livers injured by CCl4. METHOD: The adenovirus vector encoded IL-10 gene was used to transfect rats with liver injury via the caudal veins. HSC were isolated and purified from the rat livers by collagenase IV perfusion and density gradient centrifugation with Nycodenz. The expression of collagen I, IV mRNA in HSC was detected by semi-quantitative RT-PCR method and the secretion of collagen I, IV in culture serum of HSC by ELISA method. The quantity of collagen was measured in the van Gieson stained histological liver preparations. RESULTS: The expression and secretion of collagen I, IV in the adenovirus vector encoding IL-10 gene group were significantly lower than those in the adenovirus vector without IL-10 gene group and the control group (P < 0.05). The quantity of collagen in the treatment group was lower than that in the control group. CONCLUSION: IL-10 can inhibit collagen I, IV expression and secretion in rat HSC.