A blood-based three-gene signature for the non-invasive detection of early human hepatocellular carcinoma

BackgroundIdentifying early stages of disease in high-risk individuals for the development of hepatocellular carcinoma (HCC) would greatly improve the clinical outcomes of these individuals. The aim of this study was to develop a blood-based gene set that could identify early-stage HCC.MethodsComprehensive gene expression profiling of purified RNA of peripheral blood mononuclear cells (PBMC) was performed using microarrays. Gene signatures were developed through bioinformatics-driven approaches and their diagnostic value was evaluated by custom-designed, quantitative, multiplex polymerase chain reaction (PCR) assays.ResultsBioinformatics-driven analysis of microarray data derived from PBMC RNA samples of patients with HCC (N = 10), pancreatic cancer (N = 3), gastric cancer (N = 3) and 10 normal individuals identified six genes that were differentially expressed in HCC. Subsequent multiplex-PCR validation and univariate analyses performed with an independent cohort of 114 HCC patients, 48 normal individuals and 14 patients with chronic hepatitis B (CHB) validated that three genes, namely Chemokine (C-X-C motif) receptor 2 (CXCR2), C–C chemokine receptor type 2 (CCR2) and E1A-Binding Protein P400 (EP400), were able to identify HCC individually with accuracies of 82.4%, 78.4% and 65%, respectively. In combination, these three genes gave an area under the curve (AUC) of 0.96 (95% confidence interval (CI), 0.93–0.99) using multivariate logistic regression and yielded a sensitivity of 93% and a specificity of 89%. When these three genes were used in combination with alpha-fetoprotein (AFP) to predict HCC, the accuracy of predicting HCC improved slightly with an AUC of 0.99 (95% CI, 0.98–1.0), sensitivity of 93% and specificity of 95%.ConclusionsCXCR2, CCR2 and EP400 can provide a promising non-invasive multiplex PCR diagnostic assay to monitor high-risk individuals for the development of HCC.     

小扁豆凝集素结合型甲胎异质体在肝癌诊断中的意义

目的探讨小扁豆凝集素结合型甲胎蛋白异质体（AFP-L3）在良恶性肝病鉴别诊断的临床价值。方法应用装有耦联了小扁豆凝集素（LCA）的微量离心柱分离185例肝病患者的AFP-L3,用时间分辨荧光免疫检测血清AFP和AFP-L3含量,计算AFP-L3％。结果肝细胞癌患者的AFP-L3％明显高于其他肝病患者（χ^2=29．329,P〈0．001）;AFP、AFP-L3％检测肝细胞癌在ROC曲线下的面积AUC分别为0．407和0．841;以AFP-L3％≥12．6％作为诊断标准,AFP-L3诊断肝细胞癌敏感性和特异性分别为83．3％和86．3％。结论AFP-L3对肝细胞癌诊断准确度明显高于AFP,微量离心梓法柃测AFP-L3在良恶性肝脏病变鉴别诊断中具有重要临床价值。     

甲胎蛋白异质体和甲胎蛋白信使核糖核酸用于诊断肝细胞癌

目的：探讨联合检测血清肿瘤标志物甲胎蛋白（AFP）异质体（AFP—L3）和甲胎蛋白信使核糖核酸（AFPmRNA）诊断肝细胞癌及评估肝癌预后的价值。方法：对2005年4月2005年7月收治的82例肝细胞癌（HCC）、11例慢性乙型肝炎肝硬化合并门脉高压和19例肝脏良性肿瘤患者,应用酶联吸附法及荧光定量反转录一聚合酶链反应（RT—PCR）联合检测AFP—L3和AFPmRNA水平,对比其阳性率。结果：AFP—L3和AFPmRNA联合检测阳性率可达87．6％,较单独检测AFP、AFP—L3、AFP—mRNA的阳性率（分别为69．5％、81．7％、36．6％）高,差异有统计学意义;术前、术后联合检测AFP—L3和AFPmRNA均阳性患者的术后3年生存率较低。结论：术前联合检测APF-L3和AFPmRNA可提高HCC,尤其是AFP阴性HCC的诊断率,而术后联合检测对评估HCC的预后有一定的指导意义。     

Exclusion/confirmation of Ataxia-telangiectasia via cell-cycle testing

Ataxia telangiectasia (AT) is an autosomal recessive multisystem disorder with increased radiosensitivity and cancer susceptibility. The responsible gene (ATM) consists of 66 exons and a coding region of 9171 bp which precludes direct sequencing as a screening assay for confirmation or exclusion of the clinical suspicion of AT. Peripheral blood mononuclear cells of 330 patients referred for the exclusion of AT were exposed to ionizing radiation (IR) and incubated for 72 h in the presence of phytohemagglutinin. Using bivariate BrdU-Hoechst/ethidium bromide flowcytometry, the following cell cycle parameters were ascertained: (1) proportion of non-proliferating (G0,G1) cells as a measure of mitogen response, (2) proportion of first-cycle G2-phase cells relative to the growth fraction (G2/GF) as a measure of radiosensitivity. Of the cases tested, 94.2% could be unequivocally assigned either to the AT-negative or the AT-positive group of patients. Of the AT-positive cases, 11 were confirmed by ATM mutation analysis. Nineteen cases presented with non-conclusive results, mostly due to poor mitogen response; however, a combination of cell-cycle data with serum AFP concentrations led to the exclusion of AT in all but two of the uncertain cases. Substitution of ionizing radiation by the radiomimetic bleomycin was additionally tested in a small series of patients. We conclude that cell-cycle testing complemented by serum AFP measurements fulfills the criteria as a rapid and economical screening procedure for the differential diagnosis of juvenile ataxias.     

术前乳酸脱氢酶/白蛋白比值联合AFP评估肝细胞癌患者预后的价值

目的 探讨术前乳酸脱氢酶/白蛋白比值(LAR)联合甲胎蛋白(AFP)评估肝细胞癌(HCC)患者预后的价值.方法 回顾性分析106例HCC患者的临床资料;采用Kaplan-Meier法绘制生存曲线;采用单因素分析影响LAR的可能变量,应用Cox风险回归模型评估术前LAR和AFP对HCC患者预后的临床价值.结果 高LAR组...     

Decreased expression of claudin-18.2 in alpha-fetoprotein-producing gastric cancer compared to conventional gastric cancer

BackgroundAlpha-fetoprotein-producing gastric cancer (AFPGC) is a subtype of gastric cancer (GC) with more aggressive biological behavior. As a highly specific tight junction component exclusively present in gastric mucosa and gastric adenocarcinomas, claudin-18.2 (CLDN18.2) has become an emerging target in GC. In this study, we aimed to provide insight into AFPGC and investigate the expression and the clinical implications of CLDN18.2 in AFPGC.MethodsWe retrospectively collected 98 cases of AFPGC and reviewed their clinical, morphological, and immunohistochemical features. Another 356 patients with stage-matched conventional GC (cGC) were enrolled as a control group. We further surveyed CLDN18.2 expression by immunohistochemistry (IHC) in 51 AFPGC tissues and explained its association with the clinicopathological parameters of AFPGC.ResultsOur results showed that AFPGC was a unique GC type with elevated serum alpha-fetoprotein (AFP), which was a predictor of a worse prognosis. AFPGC showed typical morphological features and positive staining of at least 1 hepatocytic or enteroblastic marker. The expression rate of CLDN18.2 was low, with a positivity rate of 21.6%, which was much lower than that observed in cGC tissues (38.5%). A significant correlation was found between CLDN18.2 expression and the differentiation of AFPGC. CLDN18.2 expression was negatively correlated with the serum AFP level of AFPGC. We also found that AFPGC with a hepatoid type (HPT) component showed a significantly lower CLDN18.2 expression than those without.ConclusionsThis study demonstrated that CLDN18.2 was significantly decreased in AFPGC and was negatively correlated with the patient’s preoperative serum AFP level. The negative correlation between AFP and CLDN18.2 could be explained by retro-differentiation of AFPGC. Special treatment strategies might be needed for this unique tumor type.     

斑点免疫酶渗滤法检测血AFP

应用二个针中蛋白（ＡＦＰ）分子上不同表位的单克隆抗体个包被子硝酸纤维素膜上,另一个与辣根过氧化物酶结合,建立了检测血清中ＡＦＰ的斑点免疫酶渗滤试验。该方法简便,几分钟内即可用肉眼观察结果。该法的敏感度为３０ｎｇ／ｍｌ,以这一深度伙界,判别阳性与阴性,在４０例血清标本检测中,与ＲＩＡ法检测结果完全符合。     

肝脏硬度、层粘连蛋白及AFP值对HBV相关HCC的诊断价值

目的 评估血清甲胎蛋白（alpha-fetoprotein,AFP）、肝脏弹性（也称为硬度）测量值（liver stiffness measurement,LSM）、层粘连蛋白（laminin,LN）检测在HBV相关性肝细胞性肝癌（hepatocellular carcinoma,HCC）诊断中价值。方法 采用回顾性研究方法,纳入2013年10月至2021年3月在延安大学附属医院及延安市第二人民医院就诊的HBV感染者,行上腹部增强MRI或增强CT等检查诊断为肝硬化和肝癌的患者。检查肝纤维化四项LSM及AFP值等指标。采用统计学方法分析两组各指标的差异,Logistic单因素及多因素分析筛选诊断HCC有价值的指标。结果 两组透明质酸（hualuronic acid,HA）、Ⅳ型胶原蛋白（collagen Ⅳ,CⅣ）、Ⅲ型前胶原（procollagen type Ⅲ,PCⅢ）、LN、AFP及LSM水平差异,均有统计学意义（P均<0.05）。多因素分析显示,AFP、LN、LSM是HCC的独立危险因素（P均<0.05）。并经ROC曲线分析发现均有诊断价值,AFP的ROC曲线下面积为...     

Afp-cre-lacZ转基因小鼠的构建

目的 研究甲胎蛋白(Afp)阳性细胞的增殖分化在组织生长修复,肿瘤发生过程中的作用,建立Afp-CreERT转基因小鼠细胞示踪系统.方法 采用DNA雄原核显微注射方法获得Afp-CreERT转基因小鼠,筛选出合适的品系后,使之与Rosa26-lacZ工具小鼠杂交,获得Cre/lacZ双阳性的Afp-cre-lacZ转基因小鼠.结果 显微注射后,共出生小鼠56只,经PCR鉴定Cre阳性小鼠共4只,阳性小鼠传代后各为一系.筛选内源性Afp表达与Cre表达相对符合的品系作为Afp-CreERT转基因小鼠品系建系.Afp-CreERT转基因小鼠与Rosa26-lacZ工具小鼠杂交,经PCR鉴定后获得Cre/lacZ双阳性的Afp-cre-lacZ转基因小鼠.经实时PCR,X-gal染色,免疫荧光染色鉴定后得到Afp-cre-lacZ转基因小鼠细胞示踪系统,同时证实该系统能够正确示踪Afp表达阳性的细胞,同时也能够应用于肝损伤小鼠模型中.结论 成功构建了Afp-cre-lacZ转基因小鼠细胞示踪系统,为研究这类细胞的谱系发生提供了工具.