抗Dysbindin-1多肽双抗夹心ELISA的建立及在肝癌中的应用

目的:制备抗人Dysbindin-1特异性单克隆抗体,建立DAS-ELISA检测体系,并初步应用于肝癌血清的检测。方法:采用合成免疫原免疫BALB/c小鼠,通过杂交瘤技术制备抗Dysbindin-1单克隆抗体,用HRP标记单克隆抗体,ELISA和SDS-PAGE电泳法检测抗体的亚类、滴度;采用DAS-ELISA技术制备Dysbindin-1检测试剂盒,检测正常、肝硬化及肝癌患者各30例血清并比较其差异。结果:细胞融合后获得了4株稳定产生抗Dysbindin-1单克隆抗体的杂交瘤细胞株(1C6A11、1E8H3、2D1C11、5B6D4),抗体亚类分别为IgG2b,IgG2b,IgG1和IgG2a,杂交瘤细胞诱生的腹水抗体效价达106以上,通过抗体配对实验筛选确定2D1C11作为包被抗体,1E8H3作为酶标抗体。该ELISA方法线性范围为62.5-1000ng/mL,检测限为62.5ng/mL;应用该方法检测显示正常组与肝硬化组及肝癌组间差异显著(P0.001),Dysbindin-1诊断肝癌的敏感性和特异性分别为90%和93.3%。结论:Dysbindin-1可以成为新的候选的肝癌血清标志物,抗Dysbindin-1多肽双抗夹心ELISA体系可以初步应用于肝癌的早期诊断。     

血清AFP,CEA,CA125单独或联合检测对原发性肝癌早期诊断的临床价值研究

目的:探讨血清中甲胎蛋白(AFP)、癌胚抗原(CEA)、糖类抗原125(CA125)单独以及联合检测对于原发性肝癌的早期诊断临床价值。方法:选择2012年1月~2016年6月在我院检验科确诊的120例原发性肝癌患者作为观察组,并以80例健康志愿者作为对照组,检测和比较两组的AFP、CEA和CA125水平,分析血清AFP、CEA、CA125单项及联合检测检出原发性肝癌的阳性率和约登指数。结果:观察组血清AFP(319.53±35.78 ng/mL)、CEA(81.4±27.8 ng/mL)、CA125(20.67±4.61 ng/mL)水平均明显高于健康对照组(P0.05)。血清AFP、CEA、CA125在单独检测时诊断原发性肝癌的敏感性分别为65%(78/120)、75%(90/120)和60%(72/120),而三者的联合检测能够使检测的敏感性达到92%(112/120),显著高于单独检测时的敏感度(P0.05)。血清AFP、CEA、CA125单项检测约登指数均显著低于联合检测(P0.05)。结论:相较于血清AFP、CEA、CA125的单独检测,三者联合检测可明显提高原发性肝癌的检出率。     

不同肝病患者5种肿瘤标志物联合检测结果分析

目的探讨肿瘤标志物AFP、CEA、TSGF、CEA199和CEA125联合检测在不同肝病临床诊断中的应用价值及意义。方法选择长兴县人民医院2013年1月至2013年12月收治的36例急性乙肝、58例慢性乙肝、30例肝硬化和16例肝癌患者为研究对象,用化学发光法分别检测患者血清AFP、CEA、TSGF、CEA199和CEA125,并对各组间5种肿瘤标志物结果进行对比分析。结果肝硬化患者与对照组患者各指标相比差异有统计学意义(P0.05),肝癌患者分别与肝硬化患者和对照组患者各指标相比差异均有统计学意义(P0.01)。5种标志物联合检测能显著提高肺癌的阳性检出率(P0.05),提高检测的敏感性和特异性(P0.05)。结论 AFP、CEA、TSGF、CEA199和CEA125对肝脏良恶性疾病的诊断有一定参考价值,联合检测可以提高肝癌的检出率。     

甲胎蛋白异质体在肝细胞癌中的诊断价值分析

目的:探讨甲胎蛋白异质体(AFP-L3)的检测对肝细胞肝癌(HCC)的诊断价值。方法:选取2009年2月至2012年11月在本院治疗的90例HCC患者为研究组,另选择同期接受治疗的慢性乙型肝炎患者30例和肝炎肝硬化患者30例作为对照组。观察三组患者的AFP、AFP-L3阳性率。结果:肝癌患者AFP和AFP-L3阳性率显著高于乙肝和肝硬化患者,差异具有统计学意义(P0.05)。乙肝和肝硬化两组患者的AFP及AFP-L3阳性率无显著差异(P0.05)。AFP400 ng/m L:肝癌组AFP-L3阳性率显著高于乙肝组和肝硬化组,差异具有统计学意义(P0.05)。结论:AFP-L3在HCC中的诊断价值较甲胎蛋白大,更适合在临床诊断中应用。     

血清PIVKA-Ⅱ与AFP检测在原发性肝细胞癌诊断中的应用

为探究血清PIVKA-Ⅱ与AFP检测在原发性肝细胞癌诊断中的优劣性,对237例患者的血清PIVKA-Ⅱ与AFP进行检测,其中乙肝病毒相关性原发性肝细胞癌(hepatocellular carcinoma, HCC)患者115例、乙肝携带者(asymptomatic carrier, As C) 55例、乙肝病毒相关肝硬化(liver cirrhosis, LC)患者47例、非肝癌肿瘤患者20例。检测结果显示:肝癌组PIVKA-Ⅱ的中位表达量高于非肝癌组(包括As C组、LC组、非肝癌肿瘤组), P均小于0.05;使用AFP、PIVKA-Ⅱ和AFP+PIVKA-Ⅱ诊断肝癌的灵敏度分别为67.8%、81.7%和90.4%,对应的ROC曲线下面积为0.881、0.945和0.962, PIVKA-Ⅱ检测肝癌的cut-off值为32 m AU/m L。已有研究报道以40 m AU/m L为PIVKA-Ⅱ的cut-off值,本研究根据PIVKA-Ⅱ是否≥40 m AU/m L将HCC组分为PIVKA-Ⅱ≥40组、PIVKA-Ⅱ40组,对两组患者的性别、年龄、病毒载量、肿瘤分期、癌结节数目、肿块直径和是否抗病毒治疗进行比较,采用logistic回归分析两组患者的差异性指标,结果显示病毒载量[OR=1.150, 95%CI (1.022, 1.295), P=0.02]为PIVKA-Ⅱ检测肝癌的独立影响因素。相关分析表明PIVKA-Ⅱ与肝癌肿块直径呈正相关。此外, AFP、PIVKA-Ⅱ的cut-off值分组结果表明, PIVKA-Ⅱ≥32且AFP20组的肝癌肿块直径大于PIVKA-Ⅱ32且AFP≥20组(P=0.035)。因此, PIVKA-Ⅱ是优于AFP筛查肝细胞癌的血清学肿瘤标志物,其表达量与肿瘤肿块直径呈正相关。     

血清PIVKA-Ⅱ和AFP联合检测对肝细胞癌的临床诊断价值研究

摘要 目的：探讨血清异常凝血酶原(Protein induced by vitaminK absence or antagonist-Ⅱ,PIVKA-II)和甲胎蛋白(Alpha fetoprotein,AFP)联合检测对肝细胞癌(Hepatocellular carcinoma,HCC)的临床诊断价值。方法：选择169例HCC患者、141例肝硬化患者、66例慢性乙肝患者和100例健康体检者为研究对象,分别检测和比较其血清AFP、PIVKA-Ⅱ含量。进一步分析血清AFP、PIVKA-Ⅱ含量的相关性及其诊断HCC的敏感度、特异度、阳性预测值、阴性预测值、准确度,绘制ROC曲线,比较血清AFP、PIVKA-Ⅱ及二者联合检测诊断HCC的ROC。结果：(1)HCC组血清AFP、PIVKA-Ⅱ水平均显著高于肝硬化组、慢性乙肝组及健康对照组,差异具有统计学意义(P值均<0.05)。(2)与参考线下面积相比,AFP、PIVKA-ⅡROC曲线下面积均显著增加,差异具有统计学意义(P<0.01); 且PIVKA-Ⅱ的曲线下面积(AUC=0.790)明显大于AFP(AUC=0.708)。(3)PIVKA-Ⅱ在对HCC诊断的敏感度、特异度、阳性预测值、阴性预测值及准确度方面均优于AFP;AFP+PIVKA-Ⅱ联合检测敏感度和阴性预测值最高(P<0.05),准确度维持在75 %左右。(4)HCC患者血清AFP和PIVKA-Ⅱ水平并无显著相关性(P>0.05)。结论：血清PIVKA-Ⅱ可作为临床辅助诊断原发性肝癌的重要指标,与AFP联合检测可提高原发性肝癌的检出率。     

血清miR-21 及AFP 在大鼠肝癌发生过程中的动态表达比较分析

目的:分析血清miR-21在肝癌发生过程中的表达水平并将其与传统肝癌血清标志物甲胎蛋白(AFP)比较,探索其成为肝癌早期诊断血清标志物的可能性。方法:二乙基亚硝胺(DENA)腹腔注射诱导建立大鼠肝癌模型,建模过程中收集造癌各个阶段的血清。Realtime-PCR检测血清miR-21的表达情况,ELISA法检测血清AFP水平。结果:与正常组及纤维化期大鼠相比,miR-21在肝硬化期、肝癌早期、肝癌晚期的大鼠血清均有不同程度上调(P<0.05),AFP在肝癌早、晚期的大鼠血清明显上调(P<0.05);与肝硬化期大鼠比较,肝癌早、晚期大鼠体内的miR-21表达显著上调(P<0.05),AFP在肝癌早期、肝癌晚期均显著上调(P<0.05)。结论:血清miR-21参与了肝癌发生的过程,对于肝癌发生的各个阶段均有很大的指示作用,可能作为肝癌预防和早期诊断的一个潜在标志物。     

DEC1-mRNA在肝癌患者血清中的表达和手术预后的关系

目的:研究DEC1在肝癌(hepatocellular carcinoma,HCC)患者血清中的表达水平及其临床意义。方法:收集60例HCC患者,30例正常人,30例慢性肝炎患者,30例慢性肝硬化患者血清标本。采用实时荧光定量RT-PCR(RT-PCR)检测血清DEC1的表达情况。分析血清DEC1的表达水平与AFP相关性。探讨血清DEC1的表达水平与HCC手术预后的关系。结果:肝癌患者血清DEC1明显高于正常人、慢性肝炎和慢性肝硬化患者(P0.05)。皮尔森相关分析结果显示HCC患者血清DEC1的表达与血清AFP水平是呈正相关的。Kaplan-Meier结果显示血清DEC1低表达组术后的复发转移率显著低于高表达组,其术后生存率也显著高于高表达组(P0.01)。结论:DEC1在肝癌者血清中高表达,不仅可能成为HCC诊断的标记物,且其血清表达水平也有助于HCC临床预后的评估。     

血清Dickkopf1蛋白水平对原发性肝癌的诊断价值

目的:探索血清Dickkopf1(DKK1)水平对原发性肝癌(PLC)的诊断价值。方法:将我院收治的50例PLC患者、60例良性肝病患者、50例健康体检者及50例其它恶性肿瘤患者分别组成PLC组、良性肝病组、健康对照组及其他肿瘤组,采用ELISA定量检测各组血清DKK1蛋白的浓度,分析和比较其与α-fetoprotein(AFP)单独或联合诊断PLC的效能。结果:PLC组血清DKK1水平(838.96±104.14 ng/L)明显高于良性肝病组(322.61±25.44 ng/L)及健康对照组(213.03±25.70 ng/L),差异有统计学意义(P0.05)。AFP、DKK1单独鉴别诊断PLC和良性肝病的灵敏度分别为66%、46%,特异度分别为81.7%、96.7%,一致率分别为71.8%、73.6%;两者联合诊断的灵敏度、特异度及一致率分别为84%、78.3%、80.9%。AFP、DKK1联合诊断PLC的灵敏度和一致率均明显高于AFP、DKK1单独诊断。结论:血清DKK1在原发性肝癌中高表达,与原发性肝癌的发生密切相关,对PLC的早期诊断有一定参考价值,与AFP联合诊断可有效提高PLC的诊断效能。     

AFP、AFU、β2-MG、CA199联合检测对原发性肝癌的早期诊断价值

目的:探讨原发性肝癌患者血清甲胎蛋白(AFP)、а-L岩藻糖苷酶(AFU)、β2-微球蛋白(β2-MG)、糖类抗原-199(CA199)的含量及其联合检测对原发性肝癌的早期诊断价值。方法:选择56例原发性肝癌患者、60名肝炎肝硬化患者和60名健康对照作为研究对象,分别应用比值法和化学发光法、生化法检测其血清AFP、AFU、β2-MG、CA199的含量。结果:原发性肝癌患者血清AFP、AFU、β2-MG、CA199含量均显着高于肝炎肝硬化组及健康对照组,差异均有统计学意义(P均<0.05);联合检测AFP、AFU、β2-MG、CA199四种肿瘤标志物,其阳性率达(85.7%)明显高于AFP(53.6%)、AFU(55.4%)、β2-MG(48.2%)和CA199(42.9%)单项检测组(P均<0.05);且AFP、AFU、β2-MG、CA199四种肿瘤标志物联合检测的敏感性均高于单一检测指标,差异有统计学意义(P<0.05),但其特异性显著低于AFU、β2-MG单项检测(P<0.05)。结论:联合检测血清AFP、AFU、β2-MG、CA199含量可以提高对原发性肝癌的阳性诊断率,对诊断及鉴别诊断原发性肝癌具有重要意义。     

Association of Serum Level of Growth Differentiation Factor 15 with Liver Cirrhosis and Hepatocellular Carcinoma

Hepatocellular carcinoma (HCC) and liver cirrhosis are associated with high mortality worldwide. Currently, alpha-fetoprotein (AFP) is used as a standard serum marker for the detection of HCC, but its sensitivity and specificity are unsatisfactory, and optimal diagnostic markers for cirrhosis are lacking. We previously reported that growth differentiation factor 15 (GDF15) was significantly induced in HCV-infected hepatocytes. This study aimed to investigate GDF15 expression and its correlation with hepatitis virus-related liver diseases. A total of 412 patients with various liver diseases were studied. Healthy and Mycobacterium tuberculosis-infected subjects were included as controls. Serum and tissue GDF15 levels were measured. Serum GDF15 levels were significantly increased in patients with HCC (6.66±0.67 ng/mL, p<0.0001) and cirrhosis (6.51±1.47 ng/mL, p<0.0001) compared with healthy controls (0.31±0.01 ng/mL), though the GDF15 levels in HBV and HCV carriers were moderately elevated (1.34±0.19 ng/mL and 2.13±0.53 ng/mL, respectively). Compared with HBV or HCV carriers, GDF15 had a sensitivity of 63.1% and a specificity of 86.6% at the optimal cut-off point of 2.463 ng/mL in patients with liver cirrhosis or HCC. In HCC patients, the area under the receiver operating curve was 0.84 for GDF15 and 0.76 for AFP, but 0.91 for the combined GDF15 and AFP. Serum GDF15 levels did not significantly differ between the high-AFP and low-AFP groups. GDF15 protein expression in HCC was significantly higher than that in the corresponding adjacent paracarcinomatous tissue and normal liver. Using a combination of GDF15 and AFP will improve the sensitivity and specificity of HCC diagnosis. Further research and the clinical implementation of serum GDF15 measurement as a biomarker for HCC and cirrhosis are recommended.     

Improvement of liver cancer detection with simultaneous assessment of circulating levels of free alpha-fetoprotein (AFP) and AFP-IgM complexes

We assessed the presence of alpha-fetoprotein (AFP) complexed with IgM (AFP-IgM IC) in serum of patients affected by hepatocellular carcinoma (HCC), cirrhosis and chronic hepatitis as well as in healthy subjects by means of a dedicated ELISA assay. The amount of AFP-IgM IC was expressed in arbitrary units (AU) on a reference standard curve. Free AFP (FAFP) levels were determined in parallel in each sample by means of an automated immunoassay system. The mean serum concentration of AFP-IgM IC was significantly higher in HCC patients (mean +/- SD: 1378.3 +/- 2935.7 AU/mL) than in cirrhotic patients (129.8 +/- 261.4 AU/mL) and in patients with chronic hepatitis (80.9 +/- 168.9 AU/mL) (p < 0.01). HCC patients had FAFP values above the 20 ng/mL cutoff in 44% of cases (22/50) and AFP-IgM IC values above the 120 AU/mL cutoff in 60% of cases (30/50). The occurrence of the free and IgM-complexed form of circulating AFP did not overlap, and 82% of patients (41/50) were positive for at least one marker. The results indicate that AFP-IgM IC is a complementary serological marker to FAFP and that the combination of these biomarkers may be useful in the diagnosis of liver cancer.     

AFP, CEA, CA 19-9 and TPA in hepatocellular carcinoma.

The present study is based on the assay of four markers (AFP, CEA, TPA, Ca 19-9) using IRMA methods in 36 normal subjects, 44 cirrhosis and 66 HCC patients. Parametric and non parametric tests were used to test differences and correlations. ROC curves and discriminant functions were also elaborated. Normal 95% "cut-off" was determined by the "boostrap" method yielding: CEA 3.4 ng/ml; Ca 19-9 55 U/ml; TPA 58U/l and AFP 5.2 ng/ml. In HCC patients the values of the four markers were, on average, significantly different from those of normal subjects. However, only AFP and TPA exhibited high diagnostic accuracy (90%) for detection of the tumor. Higher than normal mean values for all markers were, also observed in cirrhotic patients. Only AFP yielded effective discrimination between HCC and cirrhosis. The positive prediction for the presence of the tumor on cirrhotic ground was 95% for AFP values higher than 18.5 ng/ml, with a 78% negative predictive value with a 6 ng/ml threshold. Association of AFP with TPA showed only a marginal diagnostic improvement. Results were not improved at all by combining CEA and Ca 19-9 with AFP and/or TPA. In conclusion, AFP is and remains the best marker for HCC and the only one effective in discriminating of HCC from cirrhosis. TPA may be considered a valid alternative if cirrhosis is not present. CEA and Ca19-9 are of no use.     

Using Serum α-Fetoprotein for Prognostic Prediction in Patients with Hepatocellular Carcinoma: What is the Most Optimal Cutoff?

Background and Aims

The prognostic ability of α-fetoprotein (AFP) for patients with hepatocellular carcinoma (HCC) was examined by using different cutoff values. The optimal AFP cutoff level is still unclear.

Methods

A total of 2579 HCC patients were consecutively enrolled in Taiwan, where hepatitis B is the major etiology of chronic liver disease. Four frequently used AFP cutoff levels, 20, 200, 400, 1000 ng/mL, were investigated. One-to-one matched pairs between patients having AFP higher and lower than the cutoffs were selected by using the propensity model. The adjusted hazard ratios of survival difference were calculated with Cox proportional hazards model.

Results

Patients with a higher AFP level were associated with more severe cirrhosis, more frequent vascular invasion, higher tumor burden and poorer performance status (all p<0.0001). In the propensity model, 4 groups of paired patients were selected, and there was no difference found in the comparison of baseline characteristics (all p>0.05). Patients with AFP <20 ng/mL had significantly better long-term survival than patients with AFP ≧20 ng/mL (p<0.0001), and patients with AFP <400 ng/mL had significantly better overall outcome than patients with AFP ≧400 ng/mL (p = 0.0186). There was no difference of long-term survival between patients divided by AFP levels of 200 and 1000 ng/mL. The adjusted hazard ratios of AFP ≧20 ng/mL and AFP ≧400 ng/mL were 1.545 and 1.471 (95% confidence interval: 1.3–1.838 and 1.178–1.837), respectively.

Conclusions

This study shows the independently predictive ability of baseline serum AFP level in HCC patients. AFP levels of 20 and 400 ng/mL are considered feasible cutoffs to predict long-term outcome in unselected HCC patients.     

Diagnostic Performance of Alpha-Fetoprotein,Protein Induced by Vitamin K Absence,Osteopontin, Dickkopf-1 and Its Combinations for Hepatocellular Carcinoma

Background & Aims

Alpha-fetoprotein (AFP) is the most widely used serum biomarker for hepatocellular carcinoma (HCC), despite its limitations. As complementary biomarkers, protein induced by vitamin K absence (PIVKA-II), osteopontin (OPN), and Dickkopf-1 (DKK-1) have been proposed. This study aimed to perform a head-to-head comparison of the diagnostic performance of AFP, PIVKA-II, OPN and DKK-1 as single or in combination to seek the best biomarker or panel, and to investigate the clinical factors affecting their performance.

Methods

Using 401 stored plasma samples obtained from 208 HCC patients and 193 liver cirrhosis control patients, plasma AFP, PIVKA-II, OPN and DKK-1 levels were measured by ELISA, and receiver operating characteristic curve analyses were performed for each biomarker and for every combination of two to four markers.

Results

Of the four biomarkers, AFP showed the highest area under the curve (0.786). The sensitivity and specificity for each single biomarker was 62% and 90.2% (AFP>20 ng/mL), 51.0% and 91.2% (PIVKA-II>10 ng/mL), 46.2% and 80.3% (OPN>100 ng/mL), and 50.0% and 80.8% (DKK-1>500 pg/mL), respectively. Among the combinations of two biomarkers, AFP>20 ng/mL or DKK-1>500 pg/mL showed the best diagnostic performance (sensitivity 78.4%, specificity 72.5%). Triple or quadruple combination did not improve the diagnostic performance further. The patient’s age, etiology and tumor invasiveness of HCC affected the performance of each marker.

Conclusions

AFP was the most useful single biomarker for HCC diagnosis, and the combined measurement of AFP and DKK-1 could maximize the diagnostic yield. Clinical decision should be based on the consideration of various factors affecting the diagnostic performance of each biomarker. Efforts to seek novel HCC biomarkers should be continued.     

Role of alpha-fetoprotein in selection of patients with hepatocellular carcinoma waiting for liver transplantation: must we reconsider it?

Background: Milan criteria (MC) represent the most commonly adopted criteria for the selection of patients with hepatocellular carcinoma (HCC) waiting for liver transplantation (LT). However, MC are exclusively based on morphological aspects. The aim of the present study was to evaluate pre-LT-detectable biological parameters, to compare them with morphological ones in terms of tumor recurrence prediction and patient survival. Methods: A cohort of 153 consecutive adult patients who underwent LT for HCC on cirrhosis from January 1999 to March 2009 was retrospectively analyzed. Results: HCC recurrence was observed in 12 patients (7.8%). At multivariate logistic regression analysis, serum alpha-fetoprotein (AFP) was the unique independent negative risk factor for the development of HCC recurrence (odds ratio 2.0, p=0.03). Adopting a cutoff value of 210 ng/mL, patients who presented serum AFP =210 ng/mL showed a 5-year survival rate of 23.3% versus 76.2% observed in patients with pre-LT serum AFP <210 ng/mL (log-rank test: <0.0001). Conclusions: In our experience, AFP was the strongest predictor of HCC recurrence, stronger than tumor morphology. AFP could ameliorate the selection of LT candidates. Further studies to evaluate the combination of morphological and biological criteria are needed.     

细胞培养结合实时荧光RT-PCR法快速检测甲肝病毒滴度的研究

目的建立一种细胞培养与实时荧光RT-PCR相结合的快速检测甲肝病毒滴度的方法。方法根据甲肝病毒(HAV)L-A-1株5'端基因组序列,设计了2条基因特异性引物及一条探针,建立实时荧光RT-PCR法,结合细胞培养检测甲肝病毒滴度,并与ELISA检测法进行比较。结果实验中建立的方法能特异检测甲肝病毒,细胞培养8d检测病毒滴度为lg107.0CCID50/mL。同一样本重复检测3次,批内样本Ct值的变异系数最大为0.89%,批间样本Ct值变异系数最大为1.66%。建立的细胞培养结合实时荧光RT-PCR法(细胞培养8 d)与细胞培养ELISA法(细胞培养28 d)检测甲肝病毒滴度结果差异无统计学意义(P0.05)。结论该方法具有快速、灵敏、特异等优点,应用于疫苗常规检测有良好前景。     

Are 90K/MAC-2BP serum levels correlated with poor prognosis in HCC patients? Preliminary results

In this study we assessed the prognostic significance of 90K/MAC-2BP serum levels in a group of 40 hepatocellular carcinoma patients. This glycoprotein is a new, interesting serum marker that reflects the immune reaction of the host against certain viral infections and tumors such as breast, ovarian and pancreatic cancer. Hepatocellular carcinoma (HCC) is one of the most widespread tumors in the world. AFP is currently the most useful marker for HCC, in spite of its poor diagnostic sensitivity. In this study 40 cirrhotic HCC patients were enrolled. The prevalence of viral hepatic infections in this group was 73% for HCV, 8% for HBV, and 8% for both viruses. Thirteen percent of the patients showed non-virus-related liver damage. 90K serum levels were assayed by an ELISA kit and AFP levels by a chemiluminescent enzyme immunometric system. The overall survival curves were estimated by the Kaplan-Meier method, taking into account age, sex, 90K and AFP serum levels. Statistical analysis showed a highly significant influence on overall survival of age below 70 years and 90K serum levels below the cutoff of 14 ng/mL. Serum AFP (< or = 20 ng/mL) had positive prognostic value only when it was associated with 90K levels (p < 0.02, log-rank).     

Immunohistochemical expression of pi class glutathione S-transferase and alpha-fetoprotein in hepatocellular carcinoma and chronic liver disease

OBJECTIVE: To determine whether tumor marker pi glutathione transferase (GST-pi) is expressed in hepatocellular carcinoma (HCC) and other chronic liver diseases and to compare its expression with that of alpha-fetoprotein (AFP). STUDY DESIGN: Samples used were formalin-fixed, paraffin-embedded liver tissues: normal (n = 3), chronic hepatitis B (n = 15), cirrhosis (n = 15) and HCC (n = 30). The expression of AFP and GST-pi was detected by using immunohistochemistry with the peroxidase-antiperoxidase method. AFP immunoreactivity was based on the cytoplasm of the hepatocytes, while GST-pi immunoreactivity was based on the nuclei of hepatocytes. RESULTS: In normal liver tissues, AFP was not expressed. However, there was strong staining of GST-pi in bile duct epithelium cells and weak staining in hepatocytes. Our results showed higher AFP immunoreactivity in cases of HCC (36.7%) as compared to cirrhosis (6.7%) and hepatitis B (0%), whereas GST-pi immunoreactivity was lower in cases of HCC (53.3%) as compared to cases of cirrhosis (100.0%) and hepatitis B (93.3%). Percent sensitivity of AFP determination for HCC was 36.7% as compared to 53.3% for GST-pi, thus making GST-pi a more sensitive marker for detection of HCC. This study showed a significant relationship between the intensity and percentage of cells stained in hepatitis B, cirrhosis and HCC for GST-pi immunoreactivity (P < .001, .001 and .05, respectively) but not for AFP (P > .05). Statistical analysis showed that there was no significant relationship between expression of AFP and GST-pi in cirrhosis and HCC cases. Hepatitis B virus infection in HCC cases showed a positive rate of 46.7%, with AFP staining positively in 42.9% of tissues and GST-pi staining positively in 57.1% of tissues. CONCLUSION: AFP is a diagnostic but rather insensitive tissue marker for HCC. However, the absence of AFP in benign chronic liver disease makes this marker useful in differentiating between HCC and other chronic liver diseases, whereas GST-pi can be used as a diagnostic marker for HCC as well as in detecting other chronic liver diseases.