Golgi protein-73:A biomarker for assessing cirrhosis and prognosis of liver disease patients

BACKGROUND Reliable biomarkers of cirrhosis,hepatocellular carcinoma(HCC),or progression of chronic liver diseases are missing.In this context,Golgi protein-73(GP73)also called Golgi phosphoprotein-2,was originally defined as a resident Golgi type II transmembrane protein expressed in epithelial cells.As a result,GP73 expression was found primarily in biliary epithelial cells,with only slight detection in hepatocytes.However,in patients with acute or chronic liver diseases and especially in HCC,the expression of GP73 is significantly up-regulated in hepatocytes.So far,few studies have assessed GP73 as a diagnostic or prognostic marker of liver fibrosis and disease progression.AIM To assess serum GP73 efficacy as a diagnostic marker of cirrhosis and/or HCC or as predictor of liver disease progression.METHODS GP73 serum levels were retrospectively determined by a novel GP73 ELISA(QUANTA Lite®GP73,Inova Diagnostics,Inc.,Research Use Only)in a large cohort of 632 consecutive patients with chronic viral and non-viral liver diseases collected from two tertiary Academic centers in Larissa,Greece(n=366)and Debrecen,Hungary(n=266).Aspartate aminotransferase(AST)/Platelets(PLT)ratio index(APRI)was also calculated at the relevant time points in all patients.Two hundred and three patients had chronic hepatitis B,183 chronic hepatitis C,198 alcoholic liver disease,28 autoimmune cholestatic liver diseases,15 autoimmune hepatitis,and 5 with other liver-related disorders.The duration of follow-up was 50(57)mo[median(interquartile range)].The development of cirrhosis,liver decompensation and/or HCC during follow-up were assessed according to internationally accepted guidelines.In particular,the surveillance for the development of HCC was performed regularly with ultrasound imaging and alpha-fetoprotein(AFP)determination every 6 mo in cirrhotic and every 12 mo in non-cirrhotic patients.RESULTS Increased serum levels of GP73(>20 units)were detected at initial evaluation in 277 out of 632 patients(43.8%).GP73-seropositivity correlated at baseline with the presence of cirrhosis(96.4%vs 51.5%,P<0.001),decompensation of cirrhosis(60.3%vs 35.5%,P<0.001),presence of HCC(18.4%vs 7.9%,P<0.001)and advanced HCC stage(52.9%vs 14.8%,P=0.002).GP73 had higher diagnostic accuracy for the presence of cirrhosis compared to APRI score[Area under the curve(AUC)(95%CI):0.909(0.885-0.934)vs 0.849(0.813-0.886),P=0.003].Combination of GP73 with APRI improved further the accuracy(AUC:0.925)compared to GP73(AUC:0.909,P=0.005)or APRI alone(AUC:0.849,P<0.001).GP73 levels were significantly higher in HCC patients compared to non-HCC[22.5(29.2)vs 16(20.3)units,P<0.001)and positively associated with BCLC stage[stage 0:13.9(10.8);stage A:17.1(16.8);stage B:19.6(22.3);stage C:32.2(30.8);stage D:45.3(86.6)units,P<0.001]and tumor dimensions[very early:13.9(10.8);intermediate:19.6(18.4);advanced:29.1(33.6)units,P=0.004].However,the discriminative ability for HCC diagnosis was relatively low[AUC(95%CI):0.623(0.570-0.675)].Kaplan-Meier analysis showed that the detection of GP73 in patients with compensated cirrhosis at baseline,was prognostic of higher rates of decompensation(P=0.036),HCC development(P=0.08),and liver-related deaths(P<0.001)during follow-up.CONCLUSION GP73 alone appears efficient for detecting cirrhosis and superior to APRI determination.In combination with APRI,its diagnostic performance can be further improved.Most importantly,the simple GP73 measurement proved promising for predicting a worse outcome of patients with both viral and nonviral chronic liver diseases.

比较两种方法检测血清高尔基体膜蛋白-73诊断肝细胞肝癌的价值

目的 探讨血清高尔基体膜蛋白-73（GP73）对肝细胞肝癌的诊断价值以及Westem blot和Elisa 2种检测方法的优劣.方法 收集人外周血血清120例,其中肝细胞肝癌、乙型病毒性肝炎、乙型肝炎肝硬化、健康志愿者各30例.应用Western blot方法和Elisa方法检测各组血清GP73的表达水平,计算并比较诊断肝细胞肝癌的受试者工作特征曲线下面积、灵敏度、特异度,并与应用电化学发光法检测的血清甲胎蛋白（AFP）进行比较.结果 Western blot和Elisa 2种方法检测肝细胞肝癌患者血清GP73的表达水平均高于其他各组,差异有统计学意义（P＜0.05）;Western blot方法和Elisa方法检测GP73诊断肝细胞肝癌的受试者工作特征曲线下面积、灵敏度、特异度均高于AFP,差异有统计学意义（P＜0.05）;与Westemblot方法相比,Elisa方法检测的GP73诊断HCC的灵敏度及特异度差异无统计学意义（P＞0.05）.结论 Western blot和Elisa 2种方法均可有效地检测人体外周血清中GP73的表达水平,但Elisa方法操作相对简单、快速,可作为血清中GP73表达水平检测的首选方法;血清GP73检测肝细胞肝癌的诊断价值优于AFP,有望成为肝细胞肝癌诊断的血清肿瘤标志物.

肝癌患者血清中GPC3和GP73含量的变化及其与血管新生、上皮间质转化的相关性

目的:研究肝癌患者血清中磷酯酰肌醇蛋白聚糖-3(GPC3)和高尔基体糖蛋白-73(GP73)含量的变化及其与血管新生、上皮间质转化的相关性。方法:选择在我院接受手术切除的肝癌患者作为肝癌组、同期体检的健康者作为对照组,测定两组受试者血清中GPC3、GP73的含量以及肝癌病灶、癌旁病灶内GPC3及GP73的含量、血管新生基因及上皮间质转化基因的表达量。结果:肝癌组患者血清中GPC3、GP73的含量显著高于对照组,肝癌病灶中GPC3、GP73的含量显著高于癌旁病灶且肝癌组患者血清中GPC3、GP73的含量与肝癌病灶中GPC3、GP73的含量呈正相关;肝癌病灶中SULT2B1、VEGF、Ang-2、Shh、Twist、N-cadherin的mRNA表达量显著高于癌旁病灶且与血清中GPC3、GP73的含量呈正相关,TUM5、Foxg1、E-cadherin的mRNA表达量显著低于癌旁病灶且与血清中GPC3、GP73的含量呈负相关。结论:肝癌患者血清中GPC3和GP73含量的升高与病灶内血管新生、上皮间质转化的加剧密切相关。

Effect of GP73 silencing on proliferation and apoptosis in hepatocellular cancer

AIM: To investigate the roles of Golgi protein (GP) 73 in the regulation of cell proliferation and apoptosis.

Dynamic expression of hepatic GP73 mRNA and protein and circulating GP73 during hepatocytes malignant transformation

Background:Hepatic Golgi protein-73(GP73)expression is related to hepatocellular carcinoma(HCC)progression.The aim of this study was to investigate the dynamic expression of GP73 mRNA and protein during hepatocytes malignant transformation.Methods:Human GP73 expressions in 88 HCC tissues and their self-control surrounding tissues were examined by immunohistochemistry,and survival time of HCC patients was evaluated by the Kaplan-Meier method.HCC model of Sprague-Dawley rats was made by diet containing 2-fluorenylacetamide.The rats were divided into the control,hepatocyte degeneration,precanceration,and HCC groups to observe GP73 protein and mRNA alterations during hepatocytes malignant transformation.Results:The GP73 expression was significantly higher in the cancerous tissues than that in the surrounding tissues,with shorter survival time,and the positive rates of GP73 protein in human HCC tissues were 53.3%at stage I,84.0%at stage II,84.6%at stage III,and 60.0%at stage IV,respectively.The positive rates of hepatic GP73 protein and mRNA in the rat models were none in the control group,66.7%and 44.4%in the hepatocytes degeneration group,88.9%and 77.8%in the hepatocytes precanceration group,and 100%in the HCC group,respectively.There was a positive correlation(r=0.91,P<0.01)between hepatic GP73 and serum GP73 during rat hepatocytes malignant transformation.Conclusions:Abnormal GP73 expression may be a sensitive and valuable biomarker in hepatocarcinogensis.

Changes of serum GPC3 and GP73 contents in patients with liver cancer and their correlation with angiogenesis and epithelial mesenchymal transition

Objective: To study the changes of serum phosphatidylinositol proteinglycan-3 (GPC3) and Golgi protein-73 (GP73) contents in patients with liver cancer and their correlation with angiogenesis and epithelial mesenchymal transition. Methods: The patients with liver cancer who underwent surgical resection in our hospital between April 2014 and October 2017 were selected as liver cancer group, healthy subjects who received physical examination during the same period were selected as control group, and the contents of GPC3 and GP73 in serum as well as the contents of GPC3 and GP73, the expression of angiogenesis genes and epithelial mesenchymal transition genes in liver cancer lesions and adjacent lesions of the two groups of subjects were determined. Results: GPC3 and GP73 contents in serum of liver cancer group were significantly higher than those of control group, GPC3 and GP73 contents in liver cancer lesions were significantly higher than those in adjacent lesions, and the GPC3 and GP73 contents in serum of liver cancer group were positively correlated with GPC3 and GP73 contents in liver cancer lesions;SULT2B1, VEGF, Ang-2, Shh, Twist and N-cadherin mRNA expression in liver cancer lesions were significantly higher those in adjacent lesions and positively correlated with the GPC3 and GP73 contents in serum whereas TUM5, Foxg1 and E-cadherin mRNA expression were significantly lower than those in adjacent lesions and negatively correlated with the GPC3 and GP73 contents in serum. Conclusion: The increase of serum GPC3 and GP73 contents in patients with liver cancer is closely related to the aggravation of angiogenesis and epithelial mesenchymal transition in the lesions.

GP73在肝细胞癌诊断中的价值

目的:探讨GP73(Golgi protein-73)在肝细胞癌诊断中的价值。方法:收集外周血血清504例,其中肝细胞癌144例、肝硬化50例、乙型病毒性肝炎100例、乙型肝炎病毒携带者84例、其他恶性肿瘤50例、肝良性肿瘤26例和健康志愿者50例;应用双抗体夹心酶联免疫定量测定方法和电化学发光法检测血清中GP73和甲胎蛋白(α-fetoprotein,AFP)的表达水平。结果:肝细胞癌组血清GP73的表达水平显著高于其他各组(P<0.05);受试者工作特征(receiver operating characteristic,ROC)曲线设定GP73临界值为64ng/mL时,GP73诊断肝细胞癌的灵敏度和特异度分别为83.3%和88.3%,显著高于AFP(72.2%和76.7%),差异有统计学意义(P<0.05);血清GP73联合AFP检测诊断肝细胞癌的灵敏度可达94.4%。结论:血清GP73的表达水平用于诊断肝细胞癌的灵敏度和特异度优于AFP,血清GP73联合AFP检测可提高肝细胞癌的诊断率。