Adenosine nucleotide translocases (ANTs) are a family of proteins abundant in the inner mitochondrial membrane, primarily responsible for shuttling ADP and ATP across the mitochondrial membrane. Additionally, ANTs are key players in balancing mitochondrial energy metabolism and regulating cell death. ANT2 isoform, highly expressed in undifferentiated and proliferating cells, is implicated in the development and drug resistance of various tumors. We conduct a detailed analysis of the potential mechanisms by which ANT2 may influence tumorigenesis and drug resistance. Notably, the significance of ANT2 extends beyond oncology, with roles in non-tumor cell processes including blood cell development, gastrointestinal motility, airway hydration, nonalcoholic fatty liver disease, obesity, chronic kidney disease, and myocardial development, making it a promising therapeutic target for multiple pathologies. To better understand the molecular mechanisms of ANT2, this review summarizes the structural properties, expression patterns, and basic functions of the ANT2 protein. In particular, we review and analyze the controversy surrounding ANT2, focusing on its role in transporting ADP/ATP across the inner mitochondrial membrane, its involvement in the composition of the mitochondrial permeability transition pore, and its participation in apoptosis.

BACKGROUND:Sterol O-acyltransferase 1 (SOAT1) is an important target in the diagnosis and treatment of liver cancer. However, the prognostic value of SOAT1 in patients with hepatocellular carcinoma (HCC) is still not clear. AIM:To investigate the correlation of SOAT1 expression with HCC, using RNA-seq and gene expression data of The Cancer Genome Atlas (TCGA)-liver hepatocellular carcinoma (LIHC) and pan-cancer. METHODS:The correlation between SOAT1 expression and HCC was analyzed. Cox hazard regression models were conducted to investigate the prognostic value of SOAT1 in HCC. Overall survival and disease-specific survival were explored based on TCGA-LIHC data. Biological processes and functional pathways mediated by SOAT1 were characterized by gene ontology (GO) analysis and the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of differentially expressed genes. In addition, the protein-protein interaction network and co-expression analyses of SOAT1 in HCC were performed to better understand the regulatory mechanisms of SOAT1 in this malignancy. RESULTS:SOAT1 and SOAT2 were highly expressed in unpaired samples, while only SOAT1 was highly expressed in paired samples. The area under the receiver operating characteristic curve of SOAT1 expression in tumor samples from LIHC patients compared with para-carcinoma tissues was 0.748, while the area under the curve of SOAT1 expression in tumor samples from LIHC patients compared with GTEx was 0.676. Patients with higher SOAT1 expression had lower survival rates. Results from GO/KEGG and gene set enrichment analyses suggested that the PI3K/AKT signaling pathway, the IL-18 signaling pathway, the calcium signaling pathway, secreted factors, the Wnt signaling pathway, the Jak/STAT signaling pathway, the MAPK family signaling pathway, and cell-cell communication were involved in such association. SOAT1 expression was positively associated with the abundance of macrophages, Th2 cells, T helper cells, CD56bright natural killer cells, and Th1 cells, and negatively linked to the abundance of Th17 cells, dendritic cells, and cytotoxic cells. CONCLUSION:Our findings demonstrate that SOAT1 may serve as a novel target for HCC treatment, which is helpful for the development of new strategies for immunotherapy and metabolic therapy.

Background and Objective:The incidence and mortality of pancreatic cancer (PC) have increased in recent years. The current status of PC diagnosis and treatment remains grim in clinical practice because the commonly used early screening tools are not sufficient. Improving the early detection of PC and strengthening standardized comprehensive treatment remain the focus of PC research. Many studies have shown that micro RNAs (miRNAs) play an important role in the occurrence, development, and treatment of PC. It is expected that miRNAs will become new molecular markers of PC.Methods:We extracted and compiled useful information from the PubMed database that met our criteria for analyzing PC diagnosis, treatment, and prognosis.Key Content and Findings:In this narrative review, we summarize the mechanism of some miRNAs in the occurrence and development of PC and review them as potential markers for the diagnosis, treatment, and prognosis of PC. The function of miRNAs in PC has great potential in studying the pathogenesis of PC. The discovery of many important oncogenic miRNAs and their downstream targets will bring new ideas and research paths for the diagnosis and targeted therapy of PC.Conclusions:MiRNAs are expected to provide novel ideas and research directions for the diagnosis and targeted treatment of PC. However, more patient data and clinical trials are needed before miRNAs can become novel molecular markers for PC.

BACKGROUND:Circulating microRNAs (miRNAs) have become increasingly appreciated in the diagnosis and prognosis of colorectal cancer (CRC). OBJECTIVE:The aim of this study was to assess the potential diagnostic and prognostic significance of serum miR-30a-5p as a potential biomarker. METHODS:The expression levels of serum miR-30a-5p were measured in 138 cases with CRC, 50 cases with benign lesions (colorectal adenoma and polyps) and 60 healthy volunteers by quantitative real time polymerase chain reaction (qRT-PCR). RESULTS:The results showed that serum miR-30a-5p levels were frequently downregulated in patients with CRC and benign lesions in comparison with normal controls. Moreover, serum miR-30a-5p levels in early-stage CRC patients were significantly increased after surgery. Receiver-operating characteristic (ROC) curve analysis demonstrated serum miR-30a-5p could well distinguish CRC patients, early-stage CRC patients from healthy controls with a relative high value of area under the curve (AUC). Furthermore, low serum miR-30a-5p expression was more frequently occurred in CRC patients with aggressive clinical variables. Additionally, CRC patients exhibiting high serum miR-30a-5p expression had significantly prolonged overall survival than those exhibiting low expression. Finally, both univariate and multivariate analyses revealed that serum miR-30a-5p expression was an independent prognostic factor for overall survival in CRC patients. CONCLUSIONS:Collectively, these findings suggested serum miR-30a-5p might act as a novel biomarker for the diagnosis and prognosis of CRC.

Objective To evaluate microRNA (miRNA,miR)-542-3p expression in glioma and its effect on cell proliferation and migration.Methods The expression of miR-542-3p in tumor tissues of 33 patients with glioma and 29 normal brain tissues of patients with traumatic brain injury was examined by real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) was used to detect.Synthesize miR-542-3p mimics and unrelated sequences (miR-542-3p NC) were transfect into human glioma cells (U87) by Lipofectamine 2000,respectively.Cell proliferation,migration and invasion ability was examined by cell counting kit-8 (CCK-8),wound healing and Transwell assay respectively.Results The relative expression levels of miR-542-3 p in glioma and normal brain tissues were 0.74 ± 0.06 and 1.06 ± 0.09,respectively.The relative expression levels of miR-542-3p in glioma tissues were significandy lower than those in normal brain tissues (t =3.03,P ＜ 0.01).Mi-542-3p mimics and microRNAs-542-3p NC were transfected into human glioma cell line U87,and had no significant effect on the proliferation of U87 cells (t =0.236,P ＞ 0.05wever,after transfection of miR-542-3p mimics into U87 cells,the migration and invasion ability of cells increased significantly.Conclusion MiR-542-3p expression in human glioma cells is decreased,and inhibite cell migration and invasion in glioma cells.

This study aims to explore the role of long noncoding RNA (lncRNA)-HOX transcript antisense intergenic RNA (HOTAIR) in the occurrence and progression of glioma. Fresh glioma and normal brain tissues were classified into a glioma group (n = 67) and a normal group (n = 64) respectively. U87 cells were assigned into the blank, sh-NC, and sh-HOTAIR groups. Quantitative real-time polymerase chain reaction (qRT-PCR) was utilized to determine HOTAIR expression. Cell proliferation, cell cycle and cell apoptosis rates were detected by cell counting kit-8 (CCK-8) and flow cytometry (FCM). Scratch test and transwell assay were conducted for cell migration and invasion. Orthotopic glioma tumor model in nude mice was established by inoculating tumor cell suspension. Hematoxylin-Eosin (HE) staining was used to observe the growth and invasion of orthotopic glioma tumors. The expression of HOTAIR and cell viability was found to be lowest in the sh-HOTAIR group among the three groups. The sh-HOTAIR group exhibited a higher apoptotic rate and lower number of cell migration compared with the blank and sh-NC groups. Additionally, the speed of wound healing was slower, the migration distance decreased and the survival time of nude mice was extended in the sh-HOTAIR compared to the other groups. Moreover, the sh-HOTAIR group demonstrated reduced lesion sizes and inflammation, no convulsions or hemiplegia and lesser number of satellite metastases. Our findings support that down-regulation of HOTAIR could inhibit cell proliferation, promote cell apoptosis as well as suppress cell invasion and migration in the progression of glioma.

The aim of the study was to explore the clinical significance of let-7 expression in hepatocellular carcinoma (HCC).A PCR array was conducted to screen for let-7 expression in early-stage HCC. Next, the deregulation of let-7 was confirmed by quantitative real-time RT-PCR (qRT-PCR) in another set of liver tissues, including normal control (NC), chronic hepatitis (CH), liver cirrhosis (LC), HCC, and adjacent nontumor (NT) tissues. In addition, as the potential target mRNA of let-7, alpha 2(I) collagen (COL1A2) mRNA was also quantified in the above liver tissues. Finally, an association study comparing let-7 and COL1A2 and their clinical significance in HCC was conducted.PCR array analysis revealed that the expression levels of let-7a/7b/7c were significantly downregulated in early-stage HCC compared to those in NT tissues. As compared to NC samples, qRT-PCR further confirmed that let-7a/7b/7c/7e were significantly upregulated in CH, LC, and NT tissues, while there were no significant differences in expression between the HCC and NC groups. Although COL1A2 may be the target mRNA of let-7, only let-7c expression was inversely correlated with COL1A2 mRNA expression in CH tissues. In HCC tissues, levels of let-7a/7b/7c/7e were positively correlated with that of COL1A2 mRNA. The clinical significance study revealed that elevated let-7a expression was significantly correlated with serosal and vein invasion, while elevated let-7c expression was significantly correlated with vein invasion and advanced TNM stage. Elevated let-7e expression was significantly correlated with vein invasion in HCC. Significantly shorter postoperative overall survival was observed in HCC patients with high let-7c expression.The results suggest that aberrant expression of let-7a/7b/7c/7e occurs in benign liver diseases and HCC. The upregulation of let-7 expression is associated with the progression and poor prognosis of HCC, and further mechanistic studies are warranted.

Cellular senescence is a key physiological barrier against tumor and represents an option for therapeutic intervention. One pivotal intracellular stimulus causing senescence is DNA damage response, while the senescence-associated heterochromatin in cancer limits the strength of the DNA damage response to endogenous genotoxic stress or DNA-damaging agents. Therefore, targeting the maintenance of compacted chromatin in cancer cells represents an optional intervention to improve the therapeutic efficacy in cancer treatment. Given a crosstalk between methionine cycle and histone methylation, we hypothesize that pharmacologically disrupting methylation potential, defined as the ratio of cellular S-adenosylmethionine to S-adenosylhomocysteine, could affect the chromatin structures in cancer cells and thus enhance their sensitivity to DNA damage response signaling. Our results showed that 3-deazaneplanocin A, a chemical inhibitor of S-adenosylhomocysteine hydrolase, elicited a typical cellular senescence in hepatoma cells. Therapy-induced senescence by 3-deazaneplanocin A was mediated through p53-p21 pathway and triggered by enhanced ataxia-telangiectasia mutated activation related to chromatin changes. In conclusion, our study demonstrated that metabolic perturbation of chromatin status in oncogene-activated cancers could be an optional intervention to sensitize DNA damage response signaling.

Background and Aims. Previous studies indicated Squamous Cell Carcinoma Antigen 1 (SCCA1) may be involved in tumorigenesis and progress of various human malignancies by inhibiting cell apoptosis and promoting cell proliferative activity. The aim of the study was to further investigate SCCA1 expression in different extent of liver diseases and evaluate the clinical significance and prognostic value in HCC.Methods. Eighty nine patient-matched tumors and peritumoral surgical specimens and 56 liver biopsies specimens from 23 patients with chronic hepatitis B (CHB), 19 with dysplastic nodule (DN), and 14 with HCC were enrolled. An additional four normal liver (NL) samples were used as controls. SCCA1 expression in liver tissue was measured by immunochemistry. Another 28 HCC specimens and paired non-tumor tissues were used for SCCA1 detection by Western blot. The prognostic value of SCCA1 expression in HCC was evaluated by the Cox proportional hazards regression model analysis.Results. Western blot analysis showed SCCA1 positive rate in HCC was higher than the matched adjacent noncancerous tissues (p <0.001). Immunohistochemistry revealed that SCCA1-positive rate increased gradually from NL, CHB, PNT to DN and HCC (p <0.05). Clinicopathological analysis showed that SCCA1 expression was positively associated with tumor differentiation (p = 0.043) and patients' Child-Pugh score (p = 0.021). The SCCA1-poistive group showed better overall survival than the negative group (p = 0.029). Importantly, SCCA1 expression was an independent prognostic factor for the overall survival of HCC patients (hazard ratio = 3.757, p <0.001).Conclusion. SCCA1 expression pattern may relate to the progression of chronic liver diseases. Furtheiniore, our study supports a potential association of negative SCCA1 expression with poor outcome in HCC. (C) 2015 IMSS. Published by Elsevier Inc.

Objective To search for the targets of microRNA (miR)-7 in the epidermal growth factor receptor (EGFR) downstream pathway,and study the internal mechanism of glioma growth inhibition by miR-7.Methods After CHG5 glioma cells were transiently transfected with miR-7 sequence,reverse transcriptase quantitative PCR (RT-qPCR) method was used to detect the gene transfection.Methyl thiazol tetrazolium (MTT) assay was used to draw cell growth curves,Transwell assay to detect cell migration,and the soft agar colony formation assay to detect tumorigenicity.Western blotting was used to detect the expression of phosphatidylinositol 3 kinase (PI3K),Raf-1,Cyclin D1,p-protein kinase B (AKT) and pmitogen extracellular kinase 1/2 (p-MEK1/2).The TargetScan and Sanger softwares were used to co-predict the potential targets of miR-7.Luciferase experiments were used to test the relationship between miR-7 and PI3K or Raf-1.Results As compared with control and scramble groups,the ability of proliferation and migration,and clone of cells in miR-7 group were obviously declined (P ＜ 0.05).Western blotting results showed that the expression of EGFR downstream members was decreased.Luciferase experiments confirmed that both PI3K and Raf-1 were the direct targets of miR-7.Conclusion miR-7 can inhibit glioma growth by simultaneously regulating PI3K/ATK and Raf/MEK/ERK pathways both in EGFR downstream pathway.

(CLDs) was tested by methylation-specific PCR (MSP). Results: In HCC, the methylation frequencies were 78.18% in APC, 63.64% in cyclin D2, 58.18% in TFPI2, 49.09% in DKK3, 49.09% in GSTP1, 47.27% in p16, 40.00% in Sigma 14-3-3, 18.18% in SFRP2, 16.36% in ppENK, 9.09% in DKK2, 7.27% in NPTX2, and 5.45% in LHX1. In CLD, the methylation frequencies were 27.78% in APC, 22.22% in cyclin D2, 7.41% in TFPI2, 3.70% in DKK3, 16.67% in GSTP1, 37.04% in p16, 37.04% in Sigma 14-3-3, 11.11% in SFRP2, 20.37% in ppENK, 7.41% in DKK2, 7.41% in NPTX2, and 9.26% in LHX1. The methylation frequencies of APC, cyclin D2, TFPI2, DKK3, and GSTP1 were higher in HCC than in CLD (P<0.01). The methylation index (MI) of the five-gene methylation profile was statistically higher in HCC (median, 0.6; IQR, 0.4-0.8) than CLD (median, 0.2; IQR, 0-0.2) (P<0.01). In HCC, MI was statistically related to the patient's age. Older patients with HCC had a higher MI. No significant correlation was observed between MI and other clinicopathological data. Moreover, MI was not related to the disease free survival and the overall survival in HCC. Conclusion: This five-gene methylation profile may be a promising biomarker for the assistant diagnosis of HCC.

Objective:This study aimed to detect the special methylation profile in peripheral blood for hepatocellular carcinoma (HCC). Methods:The methylation status of 12 tumor suppressor genes (TSGs) in the plasma of 55 HCCs and 54 chronic liver diseases (CLDs) was tested by methylation-specific PCR (MSP). Results:In HCC, the methylation frequencies were 78.18%in APC, 63.64%in cyclin D2, 58.18% in TFPI2, 49.09% in DKK3, 49.09% in GSTP1, 47.27% in p16, 40.00% in Sigma 14-3-3, 18.18% in SFRP2, 16.36% in ppENK, 9.09% in DKK2, 7.27% in NPTX2, and 5.45% in LHX1. In CLD, the methylation frequencies were 27.78% in APC, 22.22%in cyclin D2, 7.41%in TFPI2, 3.70%in DKK3, 16.67%in GSTP1, 37.04%in p16, 37.04%in Sigma 14-3-3, 11.11%in SFRP2, 20.37%in ppENK, 7.41%in DKK2, 7.41%in NPTX2, and 9.26%in LHX1. The methylation frequencies of APC, cyclin D2, TFPI2, DKK3, and GSTP1 were higher in HCC than in CLD (P<0.01). The methylation index (MI) of the five-gene methylation profile was statistically higher in HCC (median, 0.6;IQR, 0.4-0.8) than CLD (median, 0.2;IQR, 0-0.2) (P<0.01). In HCC, MI was statistically related to the patient's age. Older patients with HCC had a higher MI. No significant correlation was observed between MI and other clinicopathological data. Moreover, MI was not related to the disease free survival and the overall survival in HCC. Conclusion:This five-gene methylation profile may be a promising biomarker for the assistant diagnosis of HCC.

OBJECTIVE: To compare characters of the re-transplantation tumors from the CD133 positive and CD133 negative glioma cells in vivo.METHODS: The tissue of 26 cases of tumors from CD133 positive and 2 cases from CD133 negative glioblastoma cells were divided into four parts.The first part of tumor tissue was made to frozen section,then the percentage of apoptosis cells was detected.The second part of tumor tissue was used to extract total protein,then the expression level of Oct4,Sox2,PCNA,EGFR,Ang-2,MMP-2,MMP-9 proteins was detected by Western Blot.The third part was digested to single cell suspension,then the percentage of CD133 positive cells was detected by flow cytometry.The fourth part tumor was divided and transplated into the subcutaneous of de-thymus mice,and the tumor form rate was surveyed.RESULTS: Compared with the tumor tissue from the CD133 negative glioblastoma cells(11.14±2.15)%,the tissue from the CD133 positive glioblastoma cells had much lower percentage of apoptosis cells(4.37±0.74)%,sespectively,and higher expression level of Oct4,Sox2,PCNA,EGFR,Ang-2,MMP-2,MMP-9 proteins.The percentage of CD133 positive cells from the tumors of the CD133 positive and negative glioblastoma cells was(2.47±0.67)% and(0.44±0.14)%.The tumor forming rate from CD133 positive tumor cells was 10/10,but in CD133 negative cells that was 4/10.CONCLUSION: The cells from the tumors of the CD133 positive glioblastoma cells have more malignant performance characteristics than that from the CD133 negative cells,and both of them have higher oncogenicity than initial tumors.

BACKGROUND:Increasing evidence has shown that the deregulation of microRNAs (miRNAs) is closely related to the development and progression of hepatocellular carcinoma (HCC). To screen for HCC-specific miRNAs, this study investigated the differentially expressed miRNAs between HCC and matched non-tumorous tissue (NT). METHODS:This study analyzed the differential expression profiles of miRNAs in 11 pairs of HCC and matched NT from 11 hepatitis B virus (HBV) infection patients with the RT2 miRNA PCR array containing 88 human cancer-related miRNAs. The fold change value was more than two between the HCC and the matched NT, which indicated that there was deregulation of miRNAs. The down-regulated let-7a was validated in another sample set of 34 tissues with the TaqMan RT-qPCR method. RESULTS:Compared with the matched NT tissues, 9 miRNAs were up-regulated in the HCC tissues, and three were considered statistically significant (p < 0.05): miR-96, miR-183, and miR-196a, which were up-regulated 4.746-, 7.127-, and 3.498-fold, respectively. Simultaneously, 9 miRNAs were down-regulated in the HCC tissues, and two were considered statistically significant: let-7c and miR-138, which were down-regulated 3.945- and 4.790-fold, respectively. The expression levels of let-7a were 1.071 +/- 0.401, 0.926 +/- 0.477, 0.881 +/- 1.214, and 0.535 +/- 0.719 in the healthy group, chronic hepatitis B(CHB) group, NT group, and HCC group, respectively (p > 0.05). CONCLUSIONS:This study demonstrates that 18 miRNAs were deregulated in the HCC and matched NT tissues. The deregulated miRNAs suggest that further analyses with larger miRNA samples as a diagnostic marker are warranted.

AIM To investigate the methylation status of secreted protein acidic and rich in cysteine (SPARC) in human hepatocellular carcinoma (HCC) and evaluate its clinical implication. METHODS The methylation status of SPARC was analyzed in one HCC cell line (SMMC-7721) and 60 pairs of HCC and corresponding nontumorous tissues by methylation-specific polymerase chain reaction and bisulfite sequencing. The expression of SPARC mRNA and protein were examined by reverse transcription polymerase chain reaction and immunohistochemistry, respectively. The correlations between the methylation status and the gene expression, the clinicopathological parameters, as well as the prognosis after surgery were analyzed. RESULTS In the SMMC-7721 cell line, the loss of SPARC expression was correlated with the aberrant methylation and could be reactivated by the demethylating agent 5-aza-2'-deoxycytidine. Methylation frequency of SPARC in HCC was significantly higher than that in the corresponding nontumorous tissues (45/60 vs 7/60, P < 0.001), and it was correlated with the pathological classification (P = 0.019). The downregulation of the SPARC mRNA expression in HCC was correlated with the SPARC methylation (P = 0.040). The patients with methylated SPARC had a poorer overall survival than those without methylated SPARC (28.0 mo vs 41.0 mo, P = 0.043). CONCLUSION Aberrant methylation is an important mechanism for SPARC inactivation in HCC and SPARC methylation may be a promising biomarker for the diagnosis and prognosis of HCC.

Objective To investigate the levels of serum Golgi protein(GP73) (sGP73) in patients with HBV-related liver disease and investigate the role of sGP73 as an indicator for diagnosis of hepatocelluar carcinoma (HCC).Methods The concentration of sGP73 in patients with chronic hepatitis B (CHB,n =31),liver cirrhosis (LC,n =60),HCC (n =71),self-limited HBV infectors (n=21 ) and healthy controls (n =42) were tested by enzyme-linked immunosorbent assay (ELISA) assay and statistically analyzed in combination with relevant clinical indicators.Results The median of sGP73 in HBV-related liver disease group was significantly higher than that in the groups of self-limited HBV infectors and healthy controls respectively (P＜0.01).Among the groups with HBV-related liver disease,the median of sGP73 in LC group (231.13 ng/ml) was significantly higher than that in HCC without treatment group ( 117.63 ng/ml) (P ＜ 0.01 ) and CHB group (93.09 ng/ml) (P＜0.01).No significant difference was shown between HCC (without treatment) group and CHB group (P＞ 0.05),neither between self limited HBV infectors and healthy controls (respectively,36.79 ng/ml and 45.40 ng/ml) (P ＞ 0.05). The median of sGP73 in post-operation group (175.12 ng/ml,n=52) was significantly higher than in pre-operation HCC group (107.28 ng/ml,n=52) (P＜0.01).Along with the decreasing of liver function,sGP73 level was elevated in groups with HCC or LC.The receiver operating curve (ROC) constructed with the ratio of AFP and GP73 (AFP/GP73) showed a sensitivity of 78.87 ％ and specificity of 86.21 ％ with an area under the receiver operating curve (AUROC) of 0.878 (95％ CI:0.817-0.938) for diagnosis of HCC; comparably,a sensitivity of 67.61％ and specificity of 85.12％ were shown with a AUROC of 0.826 (95％ CI:0.755-0.897) when performed with AFP.Conclusion The level of sGP73 in HBV-related liver disease group is higher than that in the groups of self-limited HBV infector and healthy control,and it is positively correlated with the degree of hepatic impairment.For the diagnosis of HCC,joint detection of AFP and GP73 could achieve a better combination of sensitivity and specificity than the independent AFP test.

OBJECTIVE:To investigate the correlation between IL-28B rs8099917 polymorphism and the outcome of HBV infection.METHODS:Genotype of rs8099917 (T > G) in IL-28B locus was determined by TaqMan SNP genotyping from 486 individuals which including 199 chronic HBV carriers (including 100 HBV-induced liver cirrhosis and 99 HBV-related HCC). 143 people with self-limited infection and 144 healthy people served as controls. Multivariate analysis was used to assess the effect of IL-28B rs8099917 SNP among all the studied groups.RESULTS:Distribution of genotype and allele of the rs8099917 locus were in accordance with Hardy-Weinberg equilibrium in different groups or with the total population. The frequencies of the rs8099917 TT, GT, GG genotypes were 89.3%, 10.5% and 0.2%, and the frequency of allele T and G accounted for 94.5% and 5.5%, respectively. In respect of genotype or allele frequency, there was no significant differences found among the groups (P > 0.05). When comparing with the TT genotype, data from the multinomial logistic analysis showed that the ORs and (95%CI) of TG/GG genotypes were 1.589 (0.735 - 3.437), 1.351 (0.550 - 3.316) and 1.704 (0.717 - 4.052), respectively. The genotype frequencies in different groups with different clinical features showed that TG/GG genotypes significantly increased the risk of r-GTII(+) for individuals with HBV-related HCC (χ(2) = 17.534, P = 0.001), with OR as 14.821 (3.227 - 68.064). It was particularly so for males (χ(2) = 14.924, P = 0.014), with OR (95%CI) as 45.000 (2.772 - 730.571).CONCLUSION:IL-28B rs8099917 SNP had no correlation with the outcome of HBV infection.

Objective: To investigate the promoter methylation and mRNA expression of ASC gene and its clinicopathological significance in hepatocellular carcinoma (HCC). Methods: Methylation specific PCR was employed to detect the methylation status of ASC gene promoter in 58 HCC and their matched adjacent noncancerous tissue samples, 15 liver cirrhosis, 5 chronic viral hepatitis and 5 normal liver samples. Real-time fluorescent quantitation PCR was performed to examine the expression of ASC mRNA. Results: Aberrant methylation of the ASC gene was detected in 40 of 58 (69.0%) HCC tissues, and 27 of 58 (46.6%) corresponding noncancerous tissues. There was no methylation found in liver cirrhosis, chronic viral hepatitis and normal liver samples. The frequency of methylation of ASC gene promoter was higher in HCC than in adjacent noncancerous tissue (P= 0.015). ASC gene methylation was significantly associated with tumor size (P= 0.001), growth pattern (P= 0.003) and TNM staging (UICC 6th edition) (P= 0.001). Compared with those in normal liver tissue, mRNA levels were downregulated in 33 cancerous and 14 noncancerous tissue samples. ASC mRNA level was significantly lower in HCC tissue than in adjacent noncancerous tissue (P < 0.05). In 40 HCC tissue samples with ASC methylation, 26 showed low level of mRNA expression. Conclusion: The hypermethylation of ASC gene promoter is frequent and may play an important role in the development of HCC.

Background: Recent studies have focused on regulatory T cells (Tregs) in chronic hepatitis B (CHB) and hepatocellular carcinoma (HCC) and they were also conducted independently of each other. Aims: This study tried to characterize Tregs in blood and tumour infiltration, and to explore the correlations between Tregs and the context of chronic hepatitis B in HCC patients. Methods: The liver-resident Tregs and CD8 + T cells on core biopsy were investigated using immunohistochemistry staining in individuals (n = 209) with CHB (n = 47), HCC (n = 137) or healthy controls (n = 25). Circulating Tregs were detected in the above patients with CHB (n = 27) or HCC (n = 101) by flow cytometry. Results: The number of tumour-infiltrating and circulating FoxP3 + Tregs was significantly high in patients with CHB (P < 0.001). However, there were fewer intratumoural Tregs in patients with advanced HCC than those in patients with early stage HCC (P = 0.043); In contrast, the circulating Tregs frequency increased during the progression of HCC (P = 0.024). Increased tumour-infiltrating and circulating FoxP3 + Tregs were associated with poor overall survival (P = 0.041, 0.002 respectively) and a shorter time to recurrence (P = 0.049, 0.002 respectively) in patients with early stage HCC. Tumour-infiltrating Foxp3 + Tregs were related to chronic hepatitis B natural history in HCC (P = 0.012). Neither tumour-infiltrating CD8 + T cells nor balance of intratumoural Tregs and CD8 + T cells correlated with prognosis of HCC. Conclusions: Increased Foxp3 + Tregs may represent a prognostic predictor in patients with early stage HCC. The CHB natural history influenced density of tumour-infiltrating Tregs in hepatocellular carcinoma patients with chronic hepatitis B viruses infection.

Genome copy number variation (CNV) is one of the mechanisms to regulate the expression level of genes which contributes to the development and progression of cancer. In order to investigate the regions of high-level amplification and potential target genes within these amplicons in hepatocellular carcinoma (HCC), we analyzed HCC cell line (TJ3ZX-01) for CNV regions at the whole genome level using GeneChip Human Mapping 500K array, and also examined the relative copy number and expression levels of the related genes at candidate amplicons in 41 HCC tissues via real-time fluorescence quantitative PCR methods. Through analysis of sequence tag site (STS) markers by quantitative PCR, The two candidate amplicons at 1q found by SNP array were shown to occur in 56.1% (23/41) HCC samples at 1q21 and 80.5% (33/41) at 1q22–23.1. Wilcoxon signed rank test showed expression of CD1d, which located at amplicon of 1q22–23.1 increased significantly within tumor tissues compared with paired nontumor tissues. Our study provides evidences that a novel, high-level amplicon at 1q22–23.1 occurs in both HCC cell line and tissues. CD1d is a potential target for this amplicon in HCC. The up-regulation of CD1d may be used as a novel molecular signature for diagnosis and prognosis of HCC.