miR-802在原发性肝细胞癌中的表达及意义

[目的]检测微小RNA(miR)-802在原发性肝细胞癌(hepatocellular carcinoma,HCC)组织与细胞中的表达水平,分析miR-802在HCC中的临床意义。[方法]采用qRT-PCR检测HCC癌组织与癌旁组织中mi R-802的表达水平,及Hep3B、HCCLM3与HL-7702细胞的表达水平。分析miR-802表达水平与HCC临床病理特征的关系。采用Kaplan-Meier方法计算生存曲线,以Log-rank检验评估miR-802表达水平与HCC预后的关系。通过Cox比例风险回归模型单因素和多因素分析评估HCC的预后指标。[结果] HCC癌组织中miR-802的表达水平显著低于癌旁组织(P<0.05),且Hep3B、HCCLM3细胞中mi R-802的表达水平亦显著低于HL-7702细胞(P<0.05)。Ⅲ+Ⅳ期癌组织中miR-802的表达水平低于Ⅰ+Ⅱ期癌组织(P<0.05),血管转移患者癌组织中miR-802的表达水平低于非血管转移患者癌组织(P<0.05)。mi R-802低表达组患者的5年生存率显著低于高表达组(P<0.05)。单因素分析显示,低表达mi R-802、高TNM分期及阳性血管转移是HCC患者预后不良的预测指标(P<0.05)。多因素分析显示,低表达miR-802是HCC患者预后不良的唯一独立预测指标(P<0.05)。[结论] HCC组织和细胞中miR-802表达下调,低表达miR-802是HCC患者预后不良的独立预测指标。     

Tumor Associated Glycoprotein-72 is a Novel Marker for Poor Survival in Hepatocellular Carcinoma

To investigate the relationship of tumor associated glycoprotein-72 (TAG-72) expression with clinicopathological features in hepatocellular carcinoma (HCC) patients. Sixty pairs of HCC and paracarcinomatous (PCLT) tissues, and 10 nomral liver (NL) tissues were collected for Western blot analysis, and 244 pairs of HCC and PCLT tissues were collected for immunohistochemistry analysis. TAG-72 protein expression was elevated significantly in HCC tissues compared with PCLT and NL tissues. Its increased expression was correlated with TNM stage, Edmondson-Steiner grade, vein invasion and multiple tumor nodes. It is noteworthy that the HCC patients with high TAG-72 expression had shorter overall survival and disease-free survival than the patients with low expression. Multivariate Cox regression analysis revealed that TAG-72 expression was an independent prognostic factor for HCC patients. The current study demonstrated for the first time that the increased expression of TAG-72 was correlated with poor survival in patients with HCC, indicating that TAG-72 is a novel prognostic marker for HCC.     

Expression of FOXM1 and PLK1 predicts prognosis of patients with hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is one of the most frequently encountered malignant tumor types and to improve its treatment, effective prognostic biomarkers are urgently required. Cell cycle dysregulation is a significant feature of cancer progression. The aim of the present study was to estimate the expression levels of forkhead box protein M1 (FOXM1) and polo-like kinase 1 (PLK1), both of which have essential roles in cell cycle regulation, and determine their prognostic value in HCC. To this end, FOXM1 and PLK1 expression levels were assessed in The Cancer Genome Atlas and International Cancer Genome Consortium Japan HCC cohorts, and the associations between their co-expression were determined via Pearson''s correlation analysis. Furthermore, the overall survival and disease-free survival in these cohorts for different FOXM1 and PLK1 expression statuses were analyzed. In vitro knockdown experiments were also performed using Huh7 cells. The results obtained indicated overexpression of FOXM1 and PLK1 in HCC tumor tissues as well as a positive correlation between FOXM1 and PLK1 expression. The results also suggested that both FOXM1 and PLK1 are required for HCC cell proliferation. In addition, upregulation of FOXM1 and PLK1 was indicated to be associated with poor prognosis of patients with HCC. However, only their coordinated overexpression was identified as an independent prognostic factor for HCC.     

肝细胞癌组织中高尔基体糖蛋白73和分泌型聚集素的表达及其相关性分析

背景与目的：肝细胞癌(hepatocellular carcinoma,HCC)是一种具有高度侵袭、转移性的常见恶性肿瘤,预后极差。因此,研究肝癌侵袭的机制,寻找有效的干预靶点显得尤为重要。一些研究提示分泌型聚集素(secreted Clusterin,sCLU)和高尔基体糖蛋白73(Golgi glycoprotein 73,GP73)在肿瘤发生、发展及侵袭、转移过程中起重要作用,且两者具有结构和功能上的相似性。本研究旨在探讨GP73和sCLU在HCC中的表达及其与临床病理特征和预后之间的关系。方法：收集新疆医科大学第一附属医院75例HCC患者癌组织及癌旁正常肝组织标本,采用免疫组化(EnVision二步法)检测HCC组织中GP73、sCLU的表达水平;并与癌旁正常肝组织相比较;Spearman等级相关分析GP73和sCLU在肝癌中表达的关系;研究GP73和sCLU表达水平与HCC临床病理特征及预后的关系。结果：GP73和sCLU在HCC中阳性表达率分别为72%和88%,癌旁正常肝组织的阳性表达率均为4%。GP73在HCC和正常肝组织中阳性表达差异有统计学意义(χ²=73.60,P<0.05)。sCLU在HCC和正常肝组织中阳性表达差异有统计学意义(χ²=207.94,P<0.05);Spearman等级相关分析提示,HCC组织中,GP73和sCLU蛋白表达呈正相关(r=0.405,P<0.05);GP73和sCLU在HCC中表达水平与肿瘤的Edmondson病理分级、TNM分期及血管侵犯相关(P<0.05),而与患者的性别、年龄、HBsAg、合并肝硬化、AFP值、门静脉癌栓、肿瘤数目无相关性(P>0.05);GP73表达与患者生存期有关,而sCLU表达与患者生存期无关。结论：GP73和sCLU在肝癌中有较高的阳性率,且GP73和sCLU表达呈正相关,GP73和sCLU可能与HCC的侵袭性等生物行为密切相关,检测其表达将有助于评价患者预后。     

MicroRNA-212 suppresses tumor growth of human hepatocellular carcinoma by targeting FOXA1

MicroRNA-212 (miR-212) has been reported to play oncogenic or tumor suppressive role in different human malignancies. Here, we demonstrated that the mean level of miR-212 in hepatocellular carcinoma (HCC) tissues was significantly lower than that in matched tumor-adjacent tissues. Similarly, the expression of miR-212 was obviously reduced in HCC cell lines as compared with a nontransformed hepatic cell line. Ectopic expression of miR-212 inhibited cell viability and proliferation, and induced apoptosis in HepG2 cells. In contrast, down-regulation of miR-212 increased cell viability and proliferation, and suppressed apoptosis in Bel-7402 cells. In vivo studies showed that miR-212 inhibited tumor growth of HCC via suppressing proliferation and inducing apoptosis. Furthermore, we confirmed that Forkhead box protein A1 (FOXA1) was a direct target of miR-212, and it abrogated the function of miR-212 in HCC. Finally, we disclosed that the aberrant expression of miR-212 and FOXA1 was evidently correlated with poor prognostic features of HCC. MiR-212, FOXA1 and their combination were valuable prognostic markers for predicting survival of HCC patients. In conclusion, miR-212 may serve as a prognostic indicator for HCC patients and exerts tumor suppressive role, at least in part, by inhibiting FOXA1.     

Decreased expression of BATF2 is associated with a poor prognosis in hepatocellular carcinoma

Human BATF2, a basic leucine zipper protein, was recently detected in several normal immortalized cell lines but not in transformed cell lines. In addition, the expression of BATF2 also slowed the growth rate of malignant tumor cells injected into athymic nude mice. In this study, to study the role of BATF2 in hepatocellular carcinoma (HCC), we examined BATF2 expression in 50 paired HCC tumorous and nontumorous tissues, as well as in five HCC cell lines. Moreover, BATF2 expression in 114 HCC patients was evaluated using immunohistochemistry, and its relationship with clinicopathological parameters and prognosis was investigated. We found that BATF2 expression was significantly reduced in most HCC tumorous tissues, when compared with nontumorous tissues, as well as in the five HCC cell lines. Consistent with these results, the immunohistochemistry revealed that decreased BATF2 expression was present in 63 of the 114 cases and was significantly correlated with age (p = 0.006), tumor size (p = 0.046) and tumor differentiation (p = 0.030). Patients with negative BATF2 expression showed a shorter survival than those with positive expression (p = 0.016). Multivariate analysis revealed that BATF2 expression was an independent predictor of overall survival (p = 0.015). All the data support the hypothesis that BATF2 plays an important role in the progression of HCC and that it may work as a candidate tumor suppressor and a prognostic marker as well as a potential target for treatment.     

CISD2在肝细胞肝癌组织和细胞中的表达及其临床意义

目的:探究CDGSH铁硫结构域2(CISD2)在肝细胞肝癌（hepatocellular carcinoma,HCC）组织和细胞中的表达及临床病理意义。方法:采用免疫组织化学染色方法检测100例HCC肿瘤组织标本及46例癌旁肝组织标本中CISD2的蛋白表达水平,分析CISD2与临床病理特征、增殖相关蛋白Ki-67、患者生存率的相关性;运用Western blot法检测人HCC细胞株BEL7404、HepG2、SMMC-7721以及人正常肝细胞株L02中CISD2的表达,分析其表达差异。结果:CISD2在HCC肿瘤组织中的表达水平明显高于癌旁肝组织,CISD2在肝癌细胞株中的表达水平也明显高于正常肝细胞,差异有统计学意义(P＜0.01)。CISD2的表达与HCC患者的肿瘤大小（P＜0.05）以及Ki-67指数相关(P＜0.001),CISD2高表达患者的总生存率低于低表达者。结论:CISD2在HCC组织和细胞中高表达,其表达水平与患者的预后有关。CISD2可能成为HCC预后判断的标志和治疗的潜在靶点。     

A prognosis and impact factor analysis of DC-CIK cell therapy for patients with hepatocellular carcinoma undergoing postoperative TACE

Dendritic cell-cytokine-induced killer (DC-CIK) cell therapy has been experimentally implemented for enhancing anti-tumoral immunity in patients with hepatocellular carcinoma (HCC) undergoing postoperative transcatheter arterial chemoembolization (POTACE). We performed a retrospective study to evaluate the clinical efficacies of DC-CIK cell therapy and its correlations with several immune factors of the primary tumors. The overall survival time of HCC patients with HBV infection in the study group (POTACE plus DC-CIK cell therapy) was significantly longer than that of the control group (POTACE alone). The expression level of PD-L1 but not the tumor-infiltrated CD8 and CD4 T cells in the tumor tissues showed significant negative correlations with relapse-free survival (RFS) and overall survival (OS), which was also an independent prognostic factor for the five-years' suvival of patients with HCC receiving POTACE treatment. Furthermore, our study validated that PD-L1 expression was significantly inversely correlated with the survival time of HCC patients receiving POTACE plus DC-CIK cell therapy treatment. More importantly, DC-CIK cell therapy provided the best clinical benefits to HCC patients with the low PD-L1 expression receiving POTACE, which indicate that PD-L1 expression level can serve as a pivotal predictor for the therapeutic efficacy of DC-CIK cell therapy for HCC patients receiving POTACE treatment.     

Macrophage migration inhibitory factor: roles in regulating tumor cell migration and expression of angiogenic factors in hepatocellular carcinoma

Macrophage migration inhibitory factor (MIF) may contribute to multiple aspects of tumor progression, including control of cell proliferation, differentiation, cell survival and angiogenesis. However, the potential roles of MIF in regulating hepatocellular carcinoma (HCC) tumor cell migration and the expression of angiogenic factors by HCC tumor cells have not been studied yet. In our study, we reported that intracellular MIF mRNA and protein were overexpressed in HCC tissues compared to nontumor tissues by using in situ hybridization and immunohistochemic staining. HCC tumor cell lines also secreted large amounts of MIF into the supernatants of tumor cell culture. To assess the role of MIF in HCC, we employed the transwell invasion chamber to study the effect of MIF on tumor cell migration. Our results showed that recombinant MIF and the supernatants of tumor cell line culture could enhance the invasion and migration of HCC cells. This effect can be inhibited by the addition of a neutralizing anti-MIF antibody. We observed that increased MIF serum levels correlated with higher levels of interleukin-8 (IL-8) in the sera of patients with HCC than in normal volunteers. We therefore hypothesized that MIF may regulate the production of angiogenic factors by HCC cells. To test this hypothesis, we examined the effect of MIF treatment on vascular endothelial growth factor (VEGF) and IL-8 expression by HCC cell lines. MIF induced a significant dose-dependent increase in IL-8 and VEGF production. Taken together, our results indicated that MIF may act as an autocrine-acting factor that stimulates angiogenesis and metastasis in HCC by promoting expression of angiogenic factors and migration of tumor cells. A more detailed understanding of the MIF regulatory mechanisms involved may provide insight into new direction in the treatment of HCC.     

KIF21B在肝细胞癌中的表达及临床意义的研究

背景与目的:驱动蛋白家族(kinesin superfamily,KIF)是一类调控微管运动的分子马达,参与细胞运动、有丝分裂、细胞内运输及癌变过程。KIF21B作为经典的驱动蛋白分子,同时也是微管系统动态平衡的调控子,其在肿瘤中的表达尚未见报道。探讨KIF21B在肝细胞癌(hepatocellular carcinoma,HCC)中的表达水平与HCC患者临床病理学特征的关系及对预后的影响。方法:通过癌症基因组图谱(The Cancer Genome Atlas,TCGA)分析KIF21B在HCC组织及正常组织中的表达差异,及其与临床病理学特征的关系;应用实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR)检测2株HCC细胞及1株正常肝细胞中KIF21B的表达差异;应用免疫组织化学法检测于甘肃省人民医院2014年3月-2018年6月行HCC手术切除的116例HCC组织及其癌旁组织中KIF21B的表达差异,验证TCGA数据库结果,分析KIF21B表达水平与临床病理学特征之间的关系;应用COX回归模型及Kaplan-Meier法分析KIF21B表达对HCC患者预后的影响。结果:KIF21B在HCC组织的表达显著高于正常组织(P<0.01),此结果与TCGA数据库结果一致,与TNM分期、乙肝病毒感染和血管侵犯有关(P<0.05);KIF21B在2株HCC细胞中的表达明显高于正常肝细胞(P<0.05);Kaplan-Meier法分析显示,KIF21B高表达组5年生存率明显低于低表达组(P<0.05);COX分析显示,KIF21B是影响HCC预后的独立因素。结论:KIF21B在HCC组织中的表达明显高于正常组织,且与预后不良密切相关,可能参与HCC的发生、发展过程。     

S100A14 Promotes the Growth and Metastasis of Hepatocellular Carcinoma

Background: S100A14 has recently been implicated in the progress of several types of cancers. This studyaimed to investigate the clinical significance and possible mechanisms of action of S100A14 in the invasion andmetastasis of hepatocellular carcinoma (HCC). Methods: S100A14 expression in HCC was detected at mRNA andprotein levels and its prognostic significance was assessed. Functional roles of S100A14 in HCC were investigatedusing MTT, BrdU, wound healing, transwell invasion assay and HCC metastatic mouse model. Results: S100A14was significantly elevated in HCC tissues, correlated with multiple tumor nodes, high Edmondson-Steiner gradeand vascular invasion. Multivariate Cox analysis showed that the S100A14 expression level was a significant andindependent prognostic factor for overall survival (OS) of HCC patients (hazard ratio=1.98, 95% confidenceinterval=1.14-3.46, P=0.013). S100A14 promoted cell proliferation, invasion and metastasis of HCC in vitro andin vivo. Conclusion: These results suggest S100A14 is a novel prognostic marker and therapeutic target for HCC.     

Low cyclin F expression in hepatocellular carcinoma associates with poor differentiation and unfavorable prognosis

Cyclin F, capable of forming Skp1‐Cul1‐F‐box protein ubiquitin ligase complex, is implicated in controlling centrosome duplication and preventing genome instability. Cyclin F oscillates during cell cycle with a similar pattern to cyclin A. However, its expression and significance in cancer remain obscure. In this study, we showed that cyclin F was noticeably decreased in 16 pairs of tissue samples of hepatocellular carcinoma (HCC) compared to paracarcinoma tissues, at both mRNA and protein levels. Immunohistochemical staining data revealed that in 71.8% (176/245) of HCC cases, cyclin F expression in tumor tissue was much lower than that in nontumorous tissue. Low cyclin F expression, defined by receiver operating characteristic curve analysis, was present in 69.0% of HCC patients. Low expression of cyclin F was significantly correlated with tumor size, clinical stage, serum alpha‐fetoprotein level and tumor multiplicity. Further study showed that cyclin F expression was reversely associated with tumor differentiation in HCC. Kaplan – Meier analysis indicated that low cyclin F expression was related to poor overall survival and recurrence‐free survival. The prognostic impact of cyclin F was further confirmed by stratified survival analysis. Importantly, multivariate analysis revealed that low cyclin F expression was an independent poor prognostic marker for overall survival. We conclude that cyclin F is downregulated in HCC and is a promising prognostic marker for patients suffering from this deadly disease.     

Experimental and clinical evidence suggests that GRPEL2 plays an oncogenic role in HCC development

Hepatocellular carcinoma (HCC) continues to cause severe burden worldwide. The limited options especially toward HCC with metastasis prompts us to identify novel molecules for either diagnostic/prognostic or therapeutic purposes. GRPEL2 is well defined in maintaining mitochondrial homeostasis, which is critical to multiple biological processes for cancer survival. However, its role in HCC progression was not investigated before. In our analysis using data from The Cancer Genome Atlas Liver Hepatocellular Carcinoma (TCGA LIHC) dataset and tissue microarray, higher expression levels of GRPEL2 were obseved in HCC tissues compared to in normal liver tissues, and indicated higher tumor grade, higher tumor stage, and shorter overall survival (OS). Consistent with the results of above analyses, the functional experiments validated that GRPEL2 acted as a tumor-promoting factor in HCC progression. GRPEL2 knockdown suppressed cell growth, migration, and invasion in vitro, as well as inhibited tumor growth in vivo. Moreover, GRPEL2 deficiency also accelerated reactive oxygen species (ROS) production and increased mitochondrial membrane potential (MMP), leading to cell apoptosis. In addition, we found that the cell cycle and NF-κB signaling pathways were responsible for GRPEL2-induced HCC progression, based on the results of Gene Set Enrichment Analysis (GSEA) and subsequent experimental validation. Our study, for the first time, identified the role of GRPEL2 in HCC development and provided a compelling biomarker for targted therapy in HCC treatment.