HOXD10和P53蛋白在肝细胞癌组织中表达及其临床病理学意义

目的:探讨同源异型盒基因HOXD10和P53蛋白在原发性肝细胞癌hepatocellular carcinoma,HCC组织中的表达及其临床病理学意义。方法:采用免疫组化法检测62例HCC及癌旁肝组织、20例肝脏良性病变组织中HOXD10、P53蛋白的表达,并分析HCC组织中HOXD10、P53蛋白的表达与HCC患者临床病理特征的关系。结果:低分化HCC组织中HOXD10蛋白的表达显著低于中、高分化HCC、癌旁及良性病变肝组织(P0.05)。而低分化HCC组织中P53的表达显著高于中高分化HCC组织(P0.05)。HCC组织中HOXD10蛋白的表达与脉管内癌栓呈负相关(r=-0.299,P=0.026),而HCC组织中HIXD10、P53蛋白表达与患者的性别、年龄、肿瘤大小、多结节、周边肝硬化均无显著相关性(P0.05)。结论:HOXD10的低表达和P53的高表达与低分化HCC密切相关,可能作为HCC治疗和预后预测的参考指标。     

IGFBP-rP_1在原发性肝细胞肝癌中的表达及意义

目的探讨胰岛素样生长因子结合蛋白相关蛋白1(insulin-like growth factor binding proteins-related protein 1,IGFBP-rP1)在原发性肝细胞肝癌(human hepatocellar carcinomas,HCC)中的表达及意义。方法应用免疫组织化学法检测43例HCC、32例癌旁肝组织及9例正常肝组织IGFBP-rP1的表达。结果IGFBP-rP1在HCC的表达显著低于癌旁肝组织(P0.01)及正常肝组织(P0.01),但正常肝组织和癌旁肝组织IGFBP-rP1的表达无明显差异(P0.05)。IGFBP-rP1的表达强度与HCC的恶性生物学行为关系密切。结论IGFBP-rP1抑制HCC的进展,IGFBP-rP1表达下调在HCC的进展中起重要作用。     

DAP12在肝细胞癌组织中的表达及临床意义

目的:研究DAP12在肝细胞癌(HCC)组织中的表达并分析其与患者临床病理特征及预后的关系。方法:收集94例手术切除的HCC标本,采用免疫组织化学染色方法检测其DAP12、CD163的表达情况,免疫荧光共定位检测二者在HCC组织中表达的相关性。单因素统计分析DAP12与临床病理特征的关系,进一步以多因素Cox比例风险回归分析影响患者生存时间的危险因素。结果:DAP12和CD163主要表达于HCC组织内的间质细胞,且DAP12和CD163的表达位置重叠,二者表达呈显著正相关性(r=0.775,P=0.001)。DAP12表达与肿瘤直径、病理组织分级及TNM分期有关(P=0.007、0.036、0.013)。DAP12高表达组患者生存时间显著短于低表达组患者(x2=12.018,P=0.005)。DAP12高表达、TNM分期是影响患者预后的独立危险因素(P=0.013,0.039)。结论:DAP12主要在HCC组织内替代活化型巨噬细胞(TAMs)中表达,DAP12阳性TAMs浸润密度与HCC的恶性进展及预后密切相关。     

肝细胞肝癌E-Cadherin 蛋白的表达及其对判断肝癌肝移植 疗效的意义

目的:探讨E-Cadherin蛋白在肝细胞肝癌组织中的表达及其对判断肝癌肝移植患者预后的价值。方法:选择肝细胞肝癌行全肝移植患者68例,应用免疫组化方法检测其切除的肝癌组织和癌旁正常肝组织中E-Cadherin蛋白的表达情况,并对患者进行移植术后随访,分析E-Cadherin蛋白表达与肝癌肝移植患者预后的关系。结果:肝癌组织中E-Cadherin蛋白阳性表达率明显低于癌旁组织(P<0.01)。经Logrank检验分析显示,E-Cadherin蛋白低表达组移植后的无瘤生存率明显低于E-Cadherin蛋白高表达组(P<0.01)。多因素Cox回归分析显示,E-Cadherin蛋白表达是影响肝细胞肝癌患者肝移植术后肝癌复发的独立预后因素之一。结论:E-Cadherin蛋白表达是一个预测肝细胞肝癌患者肝移植预后的重要因子。     

RPN2与肝癌患者预后及对肝癌细胞生长的作用

目的:探讨核糖蛋白2(ribophorin II,RPN2)在肝细胞肝癌(HCC)组织中的表达和对HCC患者生存的影响,同时分析RPN2对肝癌HepG2细胞生长和克隆形成的作用。方法:应用免疫组化方法和HCC公共芯片数据,从蛋白和m RNA水平检测HCC组织中RPN2的表达,同时分析RPN2与HCC患者临床参数的关系及预后相关性;进一步利用MTS法和克隆形成实验在肝癌HepG2细胞中检测RPN2对细胞生长的作用。结果:98例肝癌组织中,RPN2阳性表达率88.78%,对应癌旁肝组织中,RPN2阳性表达率74.49%;癌组织中RPN2染色评分为5.80±3.15,癌旁肝组织RPN2染色评分为2.13±1.59,肝癌组织中RPN2表达显著上调(P0.001)。3个肝癌公共芯片数据(共522例肝癌)中RPN2的m RNA表达水平同样显著升高(均P0.001)。98例肝癌患者RPN2表达水平与肿瘤直径(P=0.004)、门脉侵袭(P=0.012)和TNM分期(P=0.009)相关;RPN2高表达的患者总体生存期(OS)和无复发生存期(RFS)较RPN2低表达的患者短(OS:P=0.027;RFS:P=0.036)。肝癌HepG2细胞转染RPN2小干扰RNA后,细胞生长能力显著受抑制。结论:RPN2在肝癌中表达显著升高,RPN2的表达与肝癌的恶性进展有关,RPN2显著促进肝癌细胞生长。     

肝细胞癌中NF-kBp65,VEGF和MMP-9蛋白的表达及其相关性研究

目的:探讨肝细胞癌(HCC)组织中NF-kBp65,VEGF和MMP-9的表达及其临床意义。方法:应用免疫组织化学(SP法)检测35例HCC组织,15例癌旁肝组织及10例正常肝组织中NF-kBp65,VEGF和MMP-9蛋白的表达并分析其与临床病理因素的关系。结果:NF-kBp65,VEGF和MMP-9蛋白在HCC组织中高表达率(分别为80%,74%,71%)高于对照组(P<0.01)。NF-kBp65的表达与HCC的Edmondson分级有关,即在Edmondson分级Ⅲ-Ⅳ的癌细胞中NF-kBp65的表达强于Edmondson分级Ⅰ-Ⅱ的(P<0.05);有静脉浸润的HCC组织中NF-kBp65,VEGF和MMP-9的表达高于无静脉浸润者,差异有统计学意义(P<0.05)。但NF-kBp65的表达与年龄,性别,肿瘤大小,AFP水平无关(P>0.05);VEGF和MMP-9的表达与年龄,性别,肿瘤大小,AFP水平,HCC的Edmondson分级无关(P>0.05)。NF-kBp65的表达与VEGF和MMP-9呈正相关(rs1=0.579,P1<0.05;rs2=0.406,P2<0.05)。结论:NF-kB...     

T细胞分化蛋白2在肝细胞癌中的表达及其临床意义

目的:观察T细胞分化蛋白2(Mal2)在肝细胞癌(HCC)中的表达并探讨其临床意义。方法:采用免疫组织化学染色方法检测226例HCC患者的肿瘤组织和86例正常肝组织中Mal2蛋白的表达情况,并分析其与HCC临床病理指标的关系。通过生存分析比较不同Mal2表达水平患者的生存情况,并分析影响HCC患者生存情况的危险因素。结果:Mal2蛋白在HCC肿瘤组织中的表达水平显著高于正常肝组织(P0.05)。Mal2蛋白表达水平增高与HCC患者血管侵犯、淋巴结转移和较高的TNM分期相关。生存分析表明Mal2蛋白高表达组患者术后生存率显著低于低表达组患者(P0.05)。Mal2阳性表达、血管癌栓形成、淋巴结转移及较高的TNM分期是影响HCC患者术后生存时间的独立危险因素。结论:Mal2蛋白在HCC组织中呈过表达趋势,且与肿瘤转移密切相关,可能在HCC的诊断与预后判断中有一定的应用价值。     

大肠癌组织中联合检测USP22和BMI-1表达的临床意义

目的:探讨大肠癌组织中联合检测USP22和BMI-1表达的临床意义。方法:应用定量RT-PCR检测82例大肠癌手术切除标本及相应的癌旁组织中USP22及BMI-1 mRNA的表达水平,并分析其表达的差异,研究其表达水平与大肠癌患者临床病理特征及预后的关系。结果:USP22、BMI-1 mRNA在大肠癌及癌旁组织中均可被检出,大肠癌组织中USP22、BMI-1 mRNA的相对表达水平均显著高于癌旁组织(P0.01),且二者之间呈显著正相关(r=0.708,P0.01)。USP22、BMI-1 mRNA的高表达均与大肠癌的美国癌症分期联合委员会(AJCC)分期密切相关(P0.01)。COX回归分析显示USP22及BMI-1的共表达可作为大肠癌患者预后的独立预测因素(P0.01)。结论:USP22和BMI-1的共同激活可促进大肠癌的进展,并预示预后不良。     

一种新的泛素化-铁死亡相关基因特征用于肝细胞癌患者预后分析

本研究通过筛选泛素化-铁死亡相关基因构建肝细胞癌(hepatocellular carcinoma, HCC)预后模型,并探究泛素化-铁死亡相关基因对HCC预后的影响。使用R包DESeq2对TCGA-LIHC数据集进行差异分析,获取HCC差异表达基因(HCC＿DEGs)。将HCC＿DEGs、泛素化相关基因(ubiquitination related genes, URGs)和铁死亡差异表达基因(ferroptosis＿differentially expressed genes, FERR＿EDGs)取交集获得泛素化和铁死亡相关基因(UBFE＿EDGs),将UBFE＿EDGs依次进行LASSO回归分析、单因素Cox回归分析和多因素Cox回归分析以筛选出最佳独立预后基因用于构建HCC预后模型(ubiquitination and ferroptosis-related prognostic model, UBFE)。由生存曲线、 ROC曲线和校正曲线评估UBFE的预测准确性。单因素Cox回归分析、多因素Cox回归分析评估UBFE风险评分是否为HCC的预后独立危险因素,并构建列线图模型。...     

肝细胞肝癌E-Cadherin蛋白的表达及其对判断肝癌肝移植疗效的意义

目的：探讨E-Cadherin蛋白在肝细胞肝癌组织中的表达及其对判断肝癌肝移植患者预后的价值。方法：选择肝细胞肝癌行全肝移植患者68例,应用免疫组化方法检测其切除的肝癌组织和癌旁正常肝组织中E-Cadherin蛋白的表达情况,并对患者进行移植术后随访,分析E-Cadherin蛋白表达与肝癌肝移植患者预后的关系。结果：肝癌组织中E-Cadherin蛋白阳性表达率明显低于癌旁组织（P〈0.01）。经Logrank检验分析显示,E-Cadherin蛋白低表达组移植后的无瘤生存率明显低于E-Cadherin蛋白高表达组（P〈0.01）。多因素Cox回归分析显示,E-Cadherin蛋白表达是影响肝细胞肝癌患者肝移植术后肝癌复发的独立预后因素之一。结论：E-Cadherin蛋白表达是一个预测肝细胞肝癌患者肝移植预后的重要因子。     

The expression and significance of NET-1 and Contactin in hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide and a major cause of cancer-related mortality. In this study, the significance of NET and Contactin on the pathogenesis and prognosis of HCC was investigated, and further to explore their functions in vitro and in vivo by down regulation with siRNA. The expression of NET-1 and Contactin in HCC and in adjacent non-tumor tissues (ANT) were evaluated by immunohistochemistry, and the correlations of the expression of NET-1 and Contactin with the clinicopathological characteristics and survival of HCC patients were also analyzed. After inhibited by single-target siRNA or dual-target siRNA, the expressions of NET-1 and Contactin mRNA and protein in SMMC-7221 cells were determined by RT-PCR, Western blot and immunofluorescence stain. The cell proliferation and apoptosis were assessed by CCK-8 assays and flow cytometry (FCM). The ability of cell migration and invasion were evaluated by wound-healing migrating assay and transwell chamber assays, respectively. Subsequently, transmission system encapsulated cationic liposome was used to deliver dual-siRNA into HCC xenografts in mice. The expressions of NET-1, Cortactin, Ki67, Bax, Bcl2 and Survivin in xenograft tumor were detected by immunohistochemical staining, respectively. The positive rates of NET-1 and Cortactin in HCC tissues were significantly higher than those in ANT. In HCC, the expression of NET-1 was related to Edmondson''s grade (P<0.05), cirrhosis background (P<0.001) and TNM stage (P<0.05). The expression of Cortactin was related to tumor infiltration (P<0.05), vascular invasion (P <0.001) and TNM stage (P<0.001). The expressions of NET-1 and Cortactin were positively correlated (r=0.280, P=0.004). Significant differences in the 5-year survival rates were seen between the NET-1 negative group and the positive group (P<0.05), and between the Contactin negative group (50%) and Contactin positive group (28.0%, P<0.01). The 5-year overall survival rate (OS) in NET-1 and Contactin co-expression cases (27.78%) were remarkably lower than that in both NET-1 and Contactin negative cases (54.54%) and in NET-1 positive while Contactin negative cases (47.06%, P<0.05). Univariate and multivariate Cox regression analysis revealed that NET-1 and Contactin over-expression were independent indicators for OS in HCC patients (P<0.01). There were higher expressions of NET-1 and Contactin in SMMC-7721 cells than that in other HCC cells. Dual-siRNA was demonstrated to be more effective on inhibiting cancer cell proliferation, migration and inducing apoptosis than individual siRNAs used alone in vitro and in vivo (P<0.05). The results suggest that dual-siRNA may be a great potential in siRNA-based therapeutic applications.     

Musashi2 predicts poor prognosis and invasion in hepatocellular carcinoma by driving epithelial–mesenchymal transition

The high incidence of recurrence and the poor prognosis of hepatocellular carcinoma (HCC) necessitate the discovery of new predictive markers of HCC invasion and prognosis. In this study, we evaluated the expression pattern of two members of a novel oncogene family, Musashi1 (MSI1) and Musashi2 (MSI2) in 40 normal hepatic tissue specimens, 149 HCC specimens and their adjacent non‐tumourous tissues. We observed that MSI1 and MSI2 were significantly up‐regulated in HCC tissues. High expression levels of MSI1 and MSI2 were detectable in 37.6% (56/149) and 49.0% (73/149) of the HCC specimens, respectively, but were rarely detected in adjacent non‐tumourous tissues and were never detected in normal hepatic tissue specimens. Nevertheless, only high expression of MSI2 correlated with poor prognosis. In addition, MSI2 up‐regulation correlated with clinicopathological parameters representative of highly invasive HCC. Further study indicated that MSI2 might enhance invasion of HCC by inducing epithelial–mesenchymal transition (EMT). Knockdown of MSI2 significantly decreased the invasion of HCC cells and changed the expression pattern of EMT markers. Moreover, immunohistochemistry assays of 149 HCC tissue specimens further confirmed this correlation. Taken together, the results of our study demonstrated that MSI2 correlates with EMT and has the potential to be a new predictive biomarker of HCC prognosis and invasion to help guide diagnosis and treatment of post‐operative HCC patients.     

Caveolin-1 Is Up-Regulated by GLI1 and Contributes to GLI1-Driven EMT in Hepatocellular Carcinoma

Caveolin-1 (Cav-1) has been recently identified to be over-expressed in hepatocellular carcinoma (HCC) and promote HCC cell motility and invasion ability via inducing epithelial-mesenchymal transition (EMT). However, the mechanism of aberrant overexpression of Cav-1 remains vague. Here, we observed that Cav-1 expression was positively associated with GLI1 expression in HCC tissues. Forced expression of GLI1 up-regulated Cav-1 in Huh7 cells, while knockdown of GLI1 decreased expression of Cav-1 in SNU449 cells. Additionally, silencing Cav-1 abolished GLI1-induced EMT of Huh7 cells. The correlation between GLI1 and Cav-1 was confirmed in tumor specimens from HCC patients and Cav-1 was found to be associated with poor prognosis after hepatic resection. The relationship between protein expression of GLI1 and Cav-1 was also established in HCC xenografts of nude mice. These results suggest that GLI1 may be attributed to Cav-1 up-regulation which plays an important role in GLI1-driven EMT phenotype in HCC.     

The relationship among Girdin DNA methylation,its high expression,and immune infiltration in hepatocellular carcinoma: Clues from in silico analysis

Objective: The aim of the present study was to explore the relationship among Girdin DNA methylation, its high expression, and immune infiltration in human hepatocellular carcinoma (HCC).Materials and methods: The Cancer Genome Atlas (TCGA), Gene Expression Omnibus (GEO), and International Cancer Genome Consortium (ICGC) databases were used to compare Girdin mRNA expression between HCC tissues and normal tissues, and determine the relationship between Girdin expression and HCC prognosis. TCGA database was also used to analyze the expression of Girdin and its methylation status, as well as the relationship between Girdin DNA methylation and HCC prognosis. The Tumor IMmune Estimation Resource (TIMER) database was used to explore the correlation between Girdin expression and HCC immune infiltration.Results: Girdin expression was elevated in HCC tissues compared with that in normal tissues. The degree of methylation at cg03188526, a CpG site in the Girdin gene body, was positively correlated with Girdin mRNA expression, while high Girdin expression and cg03188526 hypermethylation were both correlated with poor HCC prognosis. Additionally, HCC tissue with high Girdin expression exhibited abundant immune infiltration, and the high Girdin expression was associated with a worse prognosis in macrophage-enriched HCC specimens.Conclusion: Our findings indicated that Girdin likely functions as an oncogene in HCC and that hypermethylation at cg03188526 in the Girdin gene body may explain the high Girdin expression levels in HCC tissue. Furthermore, we report for the first time that the adverse effects of high Girdin expression in HCC patients may be partially mediated by tumor macrophage infiltration.     

MicroRNA-450b-3p inhibits cell growth by targeting phosphoglycerate kinase 1 in hepatocellular carcinoma

Dysregulation of microRNAs frequently contributes to the occurrence and progression of human diseases, including hepatocellular carcinoma (HCC). In this study, the role of miR-450b-3p in HCC was investigated. Gene Expression Omnibus database and HCC specimens were used to evaluate the expression level of miR-450b-3p and the patient's prognosis. Cell functional analyses and tumor xenograft model were used to assess the role of miR-450b-3p in HCC. Bioinformatics was used to predict the downstream target gene of miR-450b-3p, which was verified by dual-luciferase reporter assay. MiR-450b-3p was found to be downregulated in HCC cell lines and tissues, compared with nontransformed immortal hepatic cells and adjacent normal liver tissues, respectively. Lower expression of miR-450b-3p was associated with poor overall survival and disease-free survival in patients with HCC. Ectopic expression of miR-450b-3p inhibited HCC cell viability, colony formation, and cell-cycle progression in vitro, and suppressed the growth of HCC xenograft tumors in vivo. Interestingly, a negative correlation between miR-450b-3p and phosphoglycerate kinase 1 (PGK1) protein was observed among HCC specimens. Additionally, miR-450b-3p inhibited PGK1 expression and phosphorylation of protein kinase B in HCC cell lines. Further experiments confirmed that PGK1 was a direct target of miR-450b-3p. Moreover, restoration of PGK1 abrogated the inhibitory effect of miR-450b-3p on HCC proliferation and cell division. In conclusion, miR-450b-3p is downregulated in human HCC and exerts tumor suppressive effects at least in part by inhibiting PGK1.     

Decreased expression of SERPINB1 correlates with tumor invasion and poor prognosis in hepatocellular carcinoma

SERPINB1 (serine protease inhibitor, clade B, member1) is a member of the SERPINB family. Recent studies suggested that SERPINB1 may suppress the migration and invasion of lung and breast cancers. In this study, we investigated a possible involvement of SERPINB1 in the regulation of hepatocellular carcinoma metastasis (HCC). The expression of SERPINB1 was evaluated using western blot analysis in 8 paired fresh HCC specimens and immunohistochemistrical assay on 67 paraffin-embedded HCC slices. SERPINB1 was downregulated in HCC specimens and correlatively related with two clinicopathologic features of HCC, metastasis (P = 0.000) and vein invasion (P = 0.006). Univariate and multivariate survival analyses showed a lower level of SERPINB1 expression is associated with poor prognosis and clinical outcome (P = 0.001). In addition, small interfering RNA targeting SERPINB1 was used to knock down the expression of SERPINB1 in Huh7 and BEL-7404 cells. We showed that interference of SERPINB1 promoted migration and invasion of HCC cells, while cell proliferation was not affected. Finally, we observed an apparent increase in the level of active matrix metalloproteinase-2 (MMP2) after SERPINB1 knockdown, implying that SERPINB1 might participate in the regulation of HCC metastasis through modulating the activation of matrix metalloproteinases. Overall, our results suggested an inhibitory role of SERPINB1 in the migration and invasion of HCC, implying that SERPINB1 might be a potential prognostic indicator of HCC metastasis.