TNFRSF6B neutralization antibody inhibits proliferation and induces apoptosis in hepatocellular carcinoma cell

The tumor necrosis factor receptor super-family member 6b (TNFRSF6B) is over-expressed in various human cancers, but its function in hepatocellular carcinoma (HCC) remains uncertain. The aim of the study was to investigate the relationship between TNFRSF6B expression and apoptosis in HCC and the effect of anti-TNFRSF6B neutralization monoclonal antibody (McAb) on HCC cells.     

Role of hepatitis B virus genotypes and quantitative HBV DNA in metastasis and recurrence of hepatocellular carcinoma

Identification of risk factors for recurrence and metastasis of HCC is important for the prognosis of HCC surveillance in chronic HBV infection. In this article, 125 HCC patients recruited were followed up prospectively for tumor metastasis and recurrence for a median of 104 (10-130) weeks. HBV DNA level was detected by LightCycler-based real-time fluorescence quantitative polymerase chain reaction-restriction system. HBV genotypes were determined by using PCR restriction-fragment length polymorphism. BCP and PC mutations were performed by PCR and direct sequencing of amplified products. Among 125 HCC patients, 19 patients were excluded because of the lack of follow-up data and the remaining 106 patients were followed up of 2 years and entered into analysis. Sixty-nine patients had tumor metastasis or recurrence during the follow-up and the cumulative probability of HCC metastasis or recurrence was 65.1%. On multivariate analysis, genotype C and HBV DNA level were the risk factors for HCC recurrence or metastasis. The incidence of recurrence or metastasis increased with baseline HBV DNA level in a dose-response relationship ranging from 22% for HBV DNA level of less than 3 log10 copies/ml to 80% for HBV DNA level of 5 log10 copies/ml or greater (P = 0.012). Fifty-seven (74.0%) and 12 (41.4%) patients had metastasis or recurrence in patients with genotype C and B, respectively. The adjusted OR of recurrence or metastasis for genotype C compared with genotype B was 9.755 (P = 0.009). In conclusion, elevated HBV DNA level and genotype C are strong risk predictors of HCC metastasis or recurrence.     

HBx和CEACAM1在乙肝相关性肝癌中的表达及意义

目的:探讨HBx和癌胚抗原相关细胞黏附分子1(CEACAM1)在乙肝相关性肝癌中的表达及意义。方法:采用免疫组织化学方法(SP法)检测81例乙肝相关性肝癌组织中HBx及CEACAM1的表达情况,分析HBx和CEACAM1与乙肝相关性肝癌的临床病理特征。Western blot检测人正常肝细胞系QZG、肝癌细胞HepG2及稳定转染HBx的肝癌细胞HepG2(HepG2-X)中CEACAM1的表达。结果:81例乙肝相关性肝癌组织中HBx表达阳性率为74.07%(60/81),CEACAM1表达阳性率为71.60%(58/81),二者与门静脉侵袭、淋巴结转移和TNM分期存在显著相关,HBx与CEACAM1的表达呈负相关(rs=-0.310,P<0.01);在HepG2-X中,CEACAM1蛋白表达水平与肝癌细胞HepG2及人正常肝细胞系QZG相比显著降低。结论:HBx的高表达和CEACAM1的低表达与乙肝相关性肝癌的侵袭、转移密切相关,HBx有可能通过抑制CEACAM1的表达而诱导乙肝相关性肝癌的侵袭与转移。     

Hepadnaviruses in cirrhotic liver and hepatocellular carcinoma

Hepadnaviruses share properties of virion structure, genome structure and replication, epidemiologic behavior, and pathogenic effects, including an association with hepatocellular carcinoma (HCC). Epidemiologic evidence implicating hepadnavirus infection in HCC includes the observation that the geographic distributions of HBV infection and HCC are similar, that the incidence of HCC is much higher in hepadnavirus infected than uninfected hosts, and that viral DNA sequences are integrated in the cellular DNA of most (e.g., 80-90%) but not all hepadnavirus-associated HCC. Cirrhosis further increases the risk of HCC in HBV infected humans. The precise role of hepadnaviruses in development of most HCC is unclear, although the finding of viral integrations within or near protooncogenes in a few cases suggests the possibility that these integrations may play a direct role in these HCC. However, in the great majority of HCC associated with HBV infections, viral integrations are in different cellular DNA sites in different HCC, integrations are not within domains of known protooncogenes, and integrations are not found in some 10-15% hepadnavirus-associated HCC, suggesting that persisting viral sequences are not directly involved in the development of these HCC as viral sequences are for tumors caused by viruses with oncogenes or viruses that act by a "promoter-insertion" mechanism. It is possible, however, that oncogenic mutations could arise via other mutagenic mechanism that may operate in chronic hepatitis B and/or cirrhosis and which do not involve persisting viral integrations. For example, liver regeneration, which is a feature of the cirrhosis associated with chronic HBV infection (and sometimes with chronic hepatitis B) involves proliferation of many cells with HBV integrations, and such integrations have been shown to be unstable and may lead to mutations through post-integration rearrangements of cellular sequences at sites of viral integrations. Viral sequences appear to be lost or deleted at some such sites of rearranged cell DNA. Chronic HBV infection shares pathologic features of liver cell injury and reactive inflammation, liver regeneration, and in man sometimes cirrhosis with other important risk factors for HCC including chronic alcoholic liver disease, chronic non-A, non-B hepatitis, hemochromatosis, and crypogenic cirrhosis, suggesting that this common pathologic process may be carcinogenic by a mechanism that does not depend specifically on the factor which initiates liver cell injury. The pathogenetic role of chronic hepadnavirus infection in such a process would be in causing liver cell injury with reactive inflammation and hepatocyte proliferation (regeneration).(ABSTRACT TRUNCATED AT 400 WORDS)     

Integrative analysis of genome-wide association study and expression quantitative trait loci datasets identified various immune cell-related pathways for rheumatoid arthritis

Rheumatoid arthritis (RA) is an autoimmune chronic disorder manifesting as warm, swollen, and painful joints. Multiple immune cells are implicated in the development of RA. Previous studies demonstrated that integrating the genetic information of genome-wide association studies (GWAS) and expression quantitative trait loci (eQTLs) is capable of identifying new disease-risk loci and providing novel insights into the etiology of complex human disease. In this study, we conducted an integrative pathway association analysis of RA by using GWAS summary data and five immune cell types related to eQTL datasets of RA. After combining the cell-specific eQTLs and GWAS summary of RA and performing a pathway-enrichment analysis, we detected a group of RA-associated pathways with common or cell-specific enriched in the five immune cell types. 41 pathways for B cells, 33 pathways for CD4⁺ T cells, 27 pathways for CD8⁺ T cells, 39 pathways for monocyte, and 25 pathways for natural killer cells are significant in RA, among which 48% are common pathways and 32% are cell-specific pathways. We detected a group of RA-associated eQTL pathways related to five different immune cell types. Our findings may provide novel insights into the pathogenesis of RA.     

SLC4A10表达水平对肝癌患者预后的影响

目的 通过生物信息学(生信学)分析在肝细胞癌(HCC)中SLC4A10的表达对HCC患者预后的影响,初步验证并探讨其意义.方法 通过TCGA数据库,分析SLC4A10在HCC组织与正常组织间的差异表达,并利用实时定量PCR进行验证.运用Kaplan-Meier(K-M)曲线分析SLC4A10的表达量对HCC患者预后的影...     

CAS基因在原发性肝癌组织中的表达及其与HBV感染的关系

目的 探讨细胞凋亡易感基因(CAS)可否作为肝癌的病理诊断标志物,以及肝细胞癌中CAS蛋白的表达与HBV感染之间的关系.方法 应用免疫组化法检测肝癌、癌旁组织及未发生肿瘤的肝硬化、肝炎组织中CAS蛋白的表达情况,同时应用免疫组化法、核酸原位杂交法检测HBV感染的肝细胞癌组织、癌旁组织中HBsAg、HBcAg以及HBV DNA的表达情况,分析肝细胞癌中CAS蛋白的表达与HBV感染之间的关系.结果 CAS蛋白在肝癌组织中的表达较癌旁组织明显升高(P＜0.01),而癌旁组织中CAS蛋白表达较未发生肿瘤的肝硬化、肝炎组织显著增强(P＜0.01).低分化型肿瘤细胞的CAS蛋白表达明显高于中分化型和高分化型(P＜0.01).CAS蛋白在HBV感染的肝细胞癌组织中的表达明显高于非HBV感染的癌组织(P＜0.01),其中HBV DNA阳性的肝细胞癌组织中CAS蛋白表达水平显著高于HBV DNA阴性的肝细胞癌组织(P＜0.05).结论 CAS蛋白在肝细胞癌组织中呈高表达,肝细胞癌分化愈低其表达愈强,表明CAS蛋白可作为肝细胞癌的病理诊断与分化程度的评价标志物.并推测HBV DNA可能通过上调CAS的表达,在HBV感染相关性肝癌的发生发展过程中发挥重要的作用.     

乙肝病毒促进肝癌细胞系的转移侵袭力

目的探讨乙肝病毒(HBV)对肝癌细胞转移能力的影响及其可能机制。方法以初始汇合度为30%,将3种细胞系HL-7702(人正常肝细胞系)、HepG2(未转染HBV-DNA的人肝癌细胞系)、HepG2.2.15(稳定转染HBV-DNA的人肝癌细胞系)种植于96孔板中,待细胞增殖至70%汇合时,利用划痕器制造划痕伤口,置于活细胞动态成像系统中进行多时间点的显微拍照与数据采集,计算相对伤口密度(RWD),并通过免疫荧光染色与Western blot技术测定细胞中Eph A2蛋白表达,分析其与RWD值的相关性。结果细胞迁移实验中,划痕后24~96 h,HL-7702组RWD显著高于HepG2与HepG2.2.15组(P0.01),划痕后72~144 h,HepG2.2.15组RWD显著高于HepG2组(P0.01);细胞侵袭实验中,HL-7702细胞因不能穿过基质胶,而无RWD值;划痕后72~144 h,HepG2.2.15组RWD显著高于HepG2组(P0.05或P0.01)。Eph A2表达:与HL-7702组比较,HepG2与HepG2.2.15组细胞中Eph A2表达水平显著升高(P0.01),其中HepG2.2.15组中Eph A2表达水平显著高于HepG2组(P0.01),且两组肝癌细胞中Eph A2的表达量与划痕实验的RWD值呈显著正相关(迁移实验:P0.01;侵袭实验:P0.01)。结论乙肝病毒可能促进肝癌细胞的迁移和侵袭能力,其机制可能与上调Eph A2的异常表达有关。     

NF-κB在人肝细胞肝癌中的表达及与HBV X蛋白的关系

目的：研究核转录因子NF－-κB在人肝细胞肝癌组织中的表达及其与乙型肝炎病毒（HBV ）X蛋白的关系。方法；用免疫组织化学S－P法，检测52例人肝细胞肝癌组织中核转录因子NF－-κB及HBV X蛋白的表达；用脂质体介导的基因转染法将HBV x基因真核表达载体pcDNA3.1-HBX转染入人肝癌细胞系HCC－9204，检测肝癌细胞内核转录因子NF－-κB的表达。结果：52例人肝细胞肝癌组织均有核转录因子NF－-κB的广泛表达，并且在11例HBV X蛋白阳性的肝癌组织，核转录因子NF－-κB位于细胞胞质和胞核，而在41例HBV X蛋白阴性的肝癌组织，核转录因子NF－-κB位于肝癌细胞的胞质。将HBV x基因真核表达载体pcDNA3.1-HBX转染 入人肝癌细胞系HCC－9204，并在稳定表达X蛋白 的肝癌细胞，核转录因子NF－-κB定位于其胞质和胞核，而未进行基因转染的亲体细胞，核转录因子NF－-κB仅定位于细胞质，细胞核无核转录因子NF－-κB的表达。结论：核转录因子NF－-κB在人肝细胞肝癌组织中广泛表达，人肝细胞肝癌中存在着核转录因子NF－-κB的异常激活，并且核转录因子NF－-κB的异常激活与HBV X蛋白有关，X蛋白激活核转录因子NF－-κB， 使其从细胞转位于细胞核，这可能在HBV相关的人原发性肝癌肝癌的发生中起一定作用。     

Precision pathology analysis of the development and progression of hepatocellular carcinoma: Implication for precision diagnosis of hepatocellular carcinoma

Outcomes for patients with hepatocellular carcinoma (HCC) remain poor because the condition is often unresponsive to the available treatments. Consequently, the early and precise diagnosis of HCC is crucial to achieve improvements in prognosis. For patients with chronic liver disease, the assessment of liver fibrosis is also important to ascertain both the staging of fibrosis and the risk of HCC occurrence. Early HCC was first described in 1991 in Japan and was defined internationally in 2009. As the concept of early HCC spread, the multistage hepatocarcinogenesis process became accepted. Consequently, improvements in imaging technology made the early diagnosis of HCC possible. At present, the most appropriate therapeutic strategy for HCC is determined using an integrated staging system that assesses the tumor burden, the degree of liver dysfunction and the patient performance status; however, pathological and molecular features are not taken into account. The recent introduction of several new therapeutic agents will change the treatment strategy for HCC. Against this background, HCC subclassification based on tumor cellular and microenvironmental characteristics will become increasingly important. In this review, we give an overview of how pathological analysis contributes to understanding the development and progression of HCC and establishing a precision diagnosis of HCC.     

肝细胞癌组织中PEG10、SIAH2的表达及其意义

目的探讨遗传印记基因PEG10(paternally expressed gene 10,PEG10)与泛素连接酶SIAH2(seven in absentia homologs 2,SIAH2)在原发性肝细胞癌(hepatocellular carcinoma,HCC)组织中的表达及其临床意义。方法采用免疫组化SP法检测50例HCC和相应癌旁组织中PEG10、SIAH2蛋白的表达。结果 (1)PEG10和SIAH2在肝癌组织中的阳性率分别为84%和78%,高于相应癌旁组织中的阳性率8%和14%,差异有统计学意义(P<0.01)。(2)PEG10和SIAH2表达与肿瘤TNM分期有相关性(P<0.01),但与患者的年龄、性别、肿瘤大小、血清HbsAg、AFP水平、是否伴肝硬化无相关性(P>0.05)。(3)HCC中PEG10、SIAH2蛋白表达呈正相关(r=0.42,P<0.01)。结论 PEG10、SIAH2表达与HCC的发生、发展具有明显相关性。     

肿瘤易感基因101在肝细胞肝癌组织中的表达及意义

目的:检测TSG101(TSG101)在肝细胞肝癌(HCC)组织中表达的临床意义。方法:应用免疫组织化学及Westernblot方法检测TSG101蛋白在肝癌及其对应非癌肝组织组织的表达情况,并分析其在肿瘤中表达水平与患者年龄、性别、TNM分期及转移等临床病理资料之间的关系。结果:免疫组化及Western blot均显示TSG101在肝癌组织表达水平显著高于其对应非癌肝组织(P<0.05)。TSG101高表达与患者TNM分期及侵袭转移显著相关(P<0.05),而与患者性别、年龄及血清HBsAg水平无明显相关性(P>0.05)。多因素回归分析同样提示TSG101阳性表达率与患者TNM分期及转移相关(P<0.05)。结论:TSG101在肝细胞癌表达水平显著高于其对应非癌肝组织,其在肝癌表达水平与患者TNM分期及转移密切相关。     

Expression patterns and polymorphisms of PTCH in Chinese hepatocellular carcinoma patients

Aberrant activation of Hedgehog signaling pathway leads to pathological consequences in a variety of human tumors. PTCH (PTCH1), the receptor of Hedgehog pathway, is reported to function as a gatekeeper in tumor formation. Here we report, by semi-quantitative RT-PCR, PTCH expression was found in 38 hepatocellular carcinoma (HCC) patients (66%). Evidences from real time quantitative RT-PCR further indicate that compared to their matched nontumorous liver tissue, PTCH exhibit a higher expression in well and moderate differentiated tumor, but a lower expression in poorly differentiated tumor. Immunohistochemical staining showed PTCH protein was detected in the cytoplasm of 56.3% HCC samples (9/16). For the first time, we investigate the polymorphisms of PTCH in HCC. First we sequenced the recognized mutation hot spots regions of PTCH of 38 HCC samples. Two previously reported single nucleotide polymorphisms (SNPs) and a novel SNP A1056G were identified. Then we examined these three SNPs in 171 HCC samples and 162 normal liver samples. However, statistic analysis showed none of these SNPs was statistically significant for association with HCC. In conclusion, our data suggest that PTCH is involved in early stage tumor development and the Hh pathway in Chinese HCC is activated by ligand expression but not by mutation.     

Lack of association of RAD51 genetic variations with hepatitis B virus clearance and occurrence of hepatocellular carcinoma in a Korean population

The RecA homolog, E. coli (S. cerevisiae) (RAD51) may modulate hepatitis B virus (HBV) infection by maintaining genome integrity and mediating homologous DNA repairs. In this study, 16 sequence variations were detected by resequencing all exons, the exon-intron boundary, and promoter regions of the human RAD51 gene in DNA samples of 24 unrelated individuals. To investigate the association of common variations in the RAD51 locus with HBV infection and hepatocellular carcinoma (HCC) occurrence, six common polymorphisms were genotyped in a total of 1,103 Korean HBV cohort, composed of 433 spontaneously recovered patients as controls and 670 chronic carriers of HBV, who were stratified further into 327 cirrhosis/chronic hepatitis patients and 343 patients with HCC infected with HBV. Logistic analyses revealed no significant association of RAD51 polymorphisms and haplotypes with HBV clearance and HCC occurrence (P > 0.05). Furthermore, with age of infection as an important factor in disease progression to HCC, results from the Cox proportional hazards analysis showed no significant associations between any of the tested RAD51 variants and the age of onset of HCC (P > 0.05), suggesting that genetic polymorphisms of RAD51 may not play an important role in clearance of HBV and disease progression to HCC. Although studies in other populations are needed to confirm these findings, this preliminary data may contribute to the current knowledge on the pathogenesis of hepatitis.     

DNA methylation changes in normal liver tissues and hepatocellular carcinoma with different viral infection

Hepatocellular carcinoma (HCC) is known to be associated with both HBV and HCV. While epigenetic changes have been previously reported to be associated with hepatocellular carcinoma (HCC), whether the epigenetic profile of HBC associated HCC differs from that of HCV-associated HCC is unclear. We analyzed DNA methylation of ten genes (APC, CCND2, CDKN2A, GSTP1, HOXA9, RARB, RASSF1, RUNX, SFRP1, and TWIST1) using MethyLight assays on 65 archived liver tissue blocks. Three genes (APC, CCND2, and GSTP1) were frequently methylated in normal liver tissues. Five genes (APC, CDKN2A, HOXA9, RASSF1, and RUNX) were significantly more frequently methylated in malignant liver tissues than normal liver tissues. Among HCC cases, HOXA9, RASSF1 and SFRP1 were methylated more frequently in HBV-positive HCC cases, while CDKN2A were significantly more frequently methylated in HCV-positive HCC cases. Our data support the hypothesis that HCC resulting from different viral etiologies is associated with different epigenetic changes.