Ratiometric fluorescent aptasensor for determination of Golgi Protein 73 based on boron, nitrogen co-doped carbon quantum dots and copper metal-organic framework.

The aim of this study was to determine if Golgi protein-73 (GP73) is up-regulated in hepatocellular carcinoma (HCC), and to explore the possibility of using GP73 in diagnosis and treatment of HCC. Serum GP73 was detected by a quantitative ELISA assay. A total of 372 serum samples were included, among them 43 from healthy donors (Normal), 110 from either chronic hepatitis or cirrhosis (CH/LC), and 219 from HCC patients. The levels of GP73 were compared among the 3 groups. The received operating curve (ROC), sensitivity and specificity of GP73 for HCC patients were calculated. The average level of GP73 expression in normal, CH/LC and HCC groups were (22.1 ± 8.5) ng/ml, (81.4 ± 57.2) ng/ml and (271.5 ± 202.3) ng/ml, respectively. Serum GP73 levels were significantly higher in patients with HCC compared to those with CH/LC (P < 0.001). The GP73 area under ROC was 0.857. Put 100 ng/ml as the optimal cut-off point, GP73 had a sensitivity of 76.7% and a specifically of 73.2%. GP73 level had a significantly higher sensitivity than AFP (32.0%) in diagnosis of early HCC (P < 0.001). Moreover, GP73 level was elevated in the serum (72.5%, 108/149) of individuals with HCC who had serum AFP level less than 400 ng/ml. Following-up study of 4 HCC patients with low level AFP indicated that GP73 was associated with treatment and prognosis of HCC. Higher level of GP73 can be found in the serum of patients with HCC than those without. GP73 is better than AFP for the diagnosis of early HCC and in evaluating treatment result in patients with normal AFP. Further studies may help to validate both the role and mechanism of GP73 in diagnosis of HCC.

The incidence of hepatocellular carcinoma (HCC) has increased over the past decades in China. Current screening methods of HCC such as detection of α-fetoprotein (AFP) combined with liver ultrasonography remain unsatisfactory. Many HCC patients have already missed the optimal treatment period when diagnosed. Our study aimed to evaluate the value of Glypican 3 (GPC3) and Golgi protein 73 (GP73) in the detection of HCC. Thirty-nine patients with HCC and 31 patients with liver cirrhosis were enrolled. The level of serum GPC3 and GP73 were determined by ELISA. The expression of GPC3 mRNA and GP73 mRNA in peripheral blood mononuclear cell (PBMC) and liver tissues were also measured with qRT-PCR. Then, receiving operating characteristic (ROC) curves were plotted to detect the sensitivity and specificity of serum GPC3 and GP73 in the diagnosis of HCC. The levels of serum GPC3 and GP73 in the HCC group were significantly higher than in the cirrhosis group (p < 0.0001). Patients with GPC3 > 9.3 μg/L and GP73 > 77.68 ng/mL had a risk of HCC of 92.31%. The HCC diagnosis ROC curve analysis indicated that when setting the GPC3 cutoff value > 9.3 μg/L, AUC = 0.956. The sensitivity and specificity of GPC3 were 89.74% and 96.77%, respectively, with a positive predictive value of 97.2%, negative predictive value of 88.2%, + LR of 27.82 and - LR of 0.11. When setting GP73 cutoff value > 77.68 ng/mL, AUC = 0.937. The sensitivity and specificity of GP73 were 92.31% and 83.87%, respectively, with positive predictive value of 87.8%, negative predictive value of 89.7%, + LR of 5.72 and - LR of 0.092. No significant difference (p > 0.05) was found between GPC3 and GP73 AUC in ROC curves, indicating that these two biomarkers were equivalent in the prediction of HCC. The expression of serum GPC3 and GP73 was significantly higher in the HCC patients compared with the cirrhosis patients. GPC3 and GP73 might be effective non-invasive diagnostic indicators of HCC.

mTORC1 Up-Regulates GP73 to Promote Proliferation and Migration of Hepatocellular Carcinoma Cells and Growth of Xenograft Tumors in Mice

Detecting a single serum marker, such as Golgi protein 73 (GP73) or alpha-fetoprotein (AFP), may not meet the requirements for the early diagnosis of hepatocellular carcinoma (HCC) due to low sensitivity and specificity. Therefore, this study aimed to develop a simultaneous multiplex assay of GP73 and AFP. Anti-human GP73- and AFP-coupled microsphere beads and biotin-labeled detectable antibodies were prepared to develop a multiplex assay of GP73 and AFP using the Luminex xMAP technology. The assay was evaluated for cross-reactivity, standard curve, sensitivity, range of detection, and precision. Additionally, the assay was used to determine the levels of serum GP73 and AFP in healthy controls and patients with chronic hepatitis, liver cirrhosis, and HCC. The multiplex assay was successfully developed to simultaneously detect GP73 and AFP without cross-reactivity. The sensitivity for GP73 detection was 0.215 ng/mL and that for AFP detection was 0.666 ng/mL. The ranges of GP73 and AFP detection were 0.98-861.08 ng/mL and 2.01-1848.73 ng/mL, respectively. The intra- and inter-assay coefficients of variation (CVs) were <10%, indicating good precision, with recovery rates of 75-125%. The levels of serum GP73 in healthy controls, chronic hepatitis patients, liver cirrhosis patients, and HCC patients were 61.64 ± 30.60 ng/mL, 208.4 ± 99.42 ng/mL, 183.7 ± 82.78 ng/mL, and 214.1 ± 160.5 ng/mL, respectively. The levels of serum AFP in healthy controls, chronic hepatitis patients, liver cirrhosis patients, and HCC patients were 24.87 ± 14.52 ng/mL, 134.4 ± 216.5 ng/mL, 66.45 ± 133.4 ng/mL, and 891.4 ± 1278 ng/mL, respectively. The receiver operating characteristic (ROC) results showed that the area under the curves (AUC) for the combination of GP73 and AFP was 0.972, which was larger than the AUC for each marker. The sensitivity and specificity of the combined detection of GP73 and AFP for the diagnosis of HCC were 90.91% and 98.86%, respectively. The multiplex assay demonstrated a good correlation with enzyme-linked immunosorbent assay (ELISA), with correlation coefficients of 0.818 and 0.982 for GP73 (p<0.001) and AFP (p<0.001), respectively. A multiplex assay for the simultaneous detection of GP73 and AFP with high sensitivity and accuracy was developed for the diagnosis of HCC. This assay may provide a reliable reference for the early diagnosis of HCC.

Objective The aim of this work was to assess the clinical utility of Golgi protein 73 (GP73) among Egyptian hepatocellular carcinoma (HCC) patients in comparison with α-fetoprotein (AFP). In addition, the level of GP73 was evaluated after surgical resection of HCC. Background Serum Golgi protein 73 (sGP73) is a promising biomarker for detection of HCC. Patients and methods This study included 76 patients; 38 of them had proved HCC (10 of them were followed up after hepatectomy to detect the level of GP73 after treatment), 25 patients had chronic liver diseases (CLDs) and 13 apparently healthy individuals were considered as controls. Clinical examination, abdominal ultrasonography and triphasic computed tomography for focal lesion were performed. Liver function tests were performed using Synchron Cx9 ALX Clinical Autoanalyzer, serum AFP was measured using ELISA method and hepatitis markers and GP73 were determined by ELISA kit for GP73. Results There was highly statistically significant difference in GP73 between HCC and the control group and also between HCC and the CLD group. Moreover, GP73 is significantly lower after hepatectomy. For discrimination of HCC from healthy control, receiver operating characteristic curve showed that sGP73 levels had area under the receiver operating characteristic (AUROC) curve of 0.89 [95% confidence interval (CI) 0.81-0.98)] and sensitivity of 76.3% and specificity of 92.3% at a cutoff point 192 ng/l, whereas AFP had AUROC curve of 0.83 (95% CI 0.71-0.95) and sensitivity of 73.7% and specificity of 61.5% at a cutoff point 10.5 ng/ml. For discrimination of HCC from CLD, receiver operating characteristic curve showed that the sGP73 levels had the AUROC curve of 0.88 (95% CI 0.79-0.96) and sensitivity of 76.3% and specificity of 84% at a cutoff point 195 ng/l, whereas AFP had AUROC curve of 0.83 (95% CI 0.72-0.93) and sensitivity of 73.7% and specificity of 68% at a cutoff point 10.5 ng/ml. Conclusion GP73 expression is highly increased in HCC patients. Its diagnostic performance is superior to that of AFP and is a useful marker for follow-up of HCC patients after surgical hepatectomy.

BackgroundScreening of early hepatocellular carcinoma (HCC) diagnosis is the greatest challenge for hepatologists. Alpha-fetoprotein (AFP) is the most common non-invasive biomarker used in HCC diagnosis.Objectives and AimsTo make a comparison between the new biomarker Golgi protein 73 (GP73) versus the standard biomarker AFP in the diagnosis of HCC.MethodsOur study was a case–control study, and 60 patients were included in the study. They were divided into two groups: 1) HCC patients with either chronic HBV or HCV infection (n=30); and 2) non-HCC patients with HBV or HCV infection who had either chronic hepatitis or liver cirrhosis (n=30). In addition, 30 healthy volunteers were included as a control group. Patients were subjected to liver function tests, kidney function tests, serum Golgi protein 73 and AFP levels. Imaging diagnosis of HCC was done by computed tomography (CT) or magnetic resonance imaging (MRI) based on American Association for the Study of Liver Diseases (AASLD) practice guidelines.ResultsStatistically significant differences between groups in terms of serum AFP (p<0.001) and GP73 (p<0.001) were found. Non-HCC patients (chronic hepatitis and liver cirrhosis) and HCC patients had significantly higher AFP and GP73 than the control group. In addition, patients with HCC had significantly higher AFP and GP73 than chronic hepatitis and cirrhotic patients. GP73 had higher diagnostic performance than AFP. At a cut-off value of ≥8.4 ng/mL, GP73 yielded a sensitivity of 86.7% and specificity of 89% for the discrimination between HCC and normal populations. Similarly, at a cut-off value of ≥8.45 ng/mL, GP73 yielded a sensitivity of 83.3% and specificity of 84% for the discrimination between HCC patients and non-HCC patients. On the other hand, AFP at a cut-off value of ≥2.4 ng/mL yielded a sensitivity of 75.4% and specificity of 90% for the discrimination between HCC and normal populations; and at a cut-off value of ≥20.85 ng/mL, AFP yielded a sensitivity of 72.2% and specificity of 86.2% for the discrimination between HCC and non-HCC patients.ConclusionGolgi protein 73 is a promising and accurate biomarker for early detection of HCC.

To explore the sensitivity and specificity of Golgi protein 73 (GP73) monoclonal antibody in the diagnosis of hepatocellular carcinoma (HCC). Self-prepared GP73 monoclonal antibody was used as the primary antibody for detecting the serum GP73 levels in healthy controls(n=31)and HCC patients (n=59). The baseline level of the healthy controls was determined by semiquantitative analysis. The results were compared with those from GP73 polyclonal antibody and alpha-fetoprotein (AFP). The GP73 level of healthy controls was 1.2 (0.9-1.7) relative unit (RU), which was significantly lower than that of HCC patients [5.7 (2.5-7.8) RU] (P<0.001) with monoclonal antibody. Using polyclonal antibody, the GP73 level of HCC patients was also significantly higher than healthy controls [7.8 (3.0-12.4) RU vs. 1.1 (1.0-2.0) RU, P<0.001]. The sensitivity and specificity of GP73 monoclonal antibody in diagnosis of HCC were 84.7% and 93.5%; on the contrary, those of GP73 polyclonal antibody were 78.0% and 93.5%, respectively. The sensitivity and specificity of AFP (67.8% and 74.2%, respectively) in the HCC patients were markedly lower than those of GP73. Logistic regression analysis showed that the odds ratio (OR) of GP73 monoclonal antibody was 7.18 and that of GP73 polyclonal antibody was 1.51. Our self-prepared monoclonal antibody can effectively detect GP73 serum level in HCC patients, and has higher sensitivity and specificity than AFP. It may be superior to the currently used GP73 polyclonal antibody. The results lay the foundation for the further development of ELISA methods by using this monoclonal antibody.

The response of Golgi protein 73 to transcatheter arterial chemoembolization in patients with hepatocellular carcinoma may relate to the influence of certain chemotherapeutics

Background: Hepatocellular carcinoma (HCC) is amongst the most common malignant tumors that carries a poor prognosis. Clinically, alpha-fetoprotein (AFP) is the most extensively used serum biomarker for diagnosing HCC. Objectives: The current study was conducted to explore the diagnostic value of serum levels of alpha-fetoprotein-L3 (AFP-L3) and Golgi protein 73 (GP73) regarding HCC, and to determine the diagnostic accuracy of these biomarkers when used individually as well as in combination with AFP. Methodology: Blood samples were collected from 50 patients with HCV-related cirrhosis (25 subjects with HCC and 25 without HCC) recruited from the outpatient clinics of the Specialized Internal Medicine Hospital, Mansoura University, Egypt. Serum concentrations of AFP-L3 and GP73 were evaluated using enzyme-linked immunosorbent assay (ELISA). Diagnostic performance of AFP-L3 and GP73 was determined by receiver operating characteristic (ROC) curve analysis. Results: Overall, the median serum level of AFP-L3 was higher in the HCC group compared to the cirrhotic group (p=0.05). Moreover, a statistically-significant difference was observed between the median serum value of GP73 in HCC patients compared to those with cirrhosis (p &lt; 0.001). The ROC curve analysis showed that the area under the ROC curve (AUROC) values for AFP, AFP-L3 and GP73 were 0.88, 0.67 and 0.83, respectively. Of the 3 biomarkers, GP73 demonstrated the highest sensitivity (88%). The AUROC for AFP and AFP-L3 combination was 0.85, whereas that for AFP and GP73 was 0.90. Conclusion: Our findings indicate that GP73 is more sensitive than AFP and AFP-L3 in diagnosing HCC. Furthermore, the combined determination of GP73 and AFP could improve the diagnostic ability of HCC.

Simple SummaryBiliary tract cancer (BTC) represents a rare liver malignancy with unfavorable outcome. It is often challenging to identify the ideal surgical candidates and present stratification algorithms are rarely based on the individual tumor biology. In the present manuscript, we evaluated a role of serum Golgi protein 73 (GP73) in patients with resectable BTC. We could show that elevated levels of GP73 before surgery identified a subgroup of BTC patients with a significantly reduced overall survival after tumor resection. Therefore, measurement of GP73 serum levels might become a novel tool in the challenging preoperative stratification process of patients with resectable BTC.Background: Tumor resection represents the only potentially curative therapy for patients with biliary tract cancer. Nevertheless, disease recurrence is observed in about 50% of patients, leading to a 5-years survival rate of less than 50%. The Golgi protein 73 (GP73), a type II Golgi transmembrane protein, exerts important functions of intracellular protein processing and transportation. Circulating GP73 has recently been suggested as a prognostic marker following resection of hepatocellular carcinoma (HCC) but its role in the context of BTC has remained unknown. In this study, we evaluate a potential role of circulating GP73 as a novel biomarker in patients with resectable BTC. Methods: GP73 serum levels were measured by immunoassay in n = 97 BTC and n = 40 HCC patients as well as n = 31 healthy controls. Results were correlated with clinical data. Results: Serum GP73 levels were significantly elevated in BTC patients compared to healthy controls but lower compared to HCC patients. The combination of GP73/CA19-9 showed a sensitivity and specificity of 83.5% and 90.3% regarding the differentiation of BTC patients and healthy controls. BTC patients with baseline GP73 levels above the ideal cut-off value (42.47 ng/mL) showed a significantly reduced median overall survival (193 days) compared to patients with preoperative GP73 levels below this cut-off (882 days). These results were confirmed in uni- and multivariate Cox-regression analysis including several clinicopathological parameters such as age, ECOG performance status, tumor stage as well as established tumor markers and parameters of liver and kidney function. Conclusions: GP73 represents a previously unrecognized biomarker in the patients with resectable BTC that identifies patients with an impaired postoperative outcome. If larger clinical trials confirmed these findings, measurement of GP73 serum levels might become a novel tool in the challenging preoperative stratification process of patients with resectable BTC.

Development of an alpha-fetoprotein and Golgi protein 73 multiplex detection assay using xMAP technology

Hepatocellular carcinoma (HCC) incidence is fast-growing especially in countries highly prevalent with viral hepatitis. Its poor prognosis has driven the research toward the discovery of sensitive markers for early detection. We investigated the usefulness of serum Transforming growth factor-beta1 (TGF-β1), Glypican-3 (GPC3), and Golgi protein-73 (GP73) mRNAs as early biomarkers in HCC Egyptian patients chronically infected with hepatitis C virus (HCV) in comparison with serum alpha-fetoprotein (AFP). Using semi-quantitative RT-PCR and densitometry analysis, circulating TGF-β1, GPC3, and GP73 mRNAs expressions were estimated in 15 healthy adults, 15 chronic HCV (CHC) patients and 25 HCC patients. Serum GP73 expression percentage in HCC group was significantly higher than controls (100 vs. 40%, P≤0.001) and when compared to elevated serum AFP levels (100 vs. 36%, P≤0.001). TGF-β1 and GP73 expression means were also higher in HCC patients than controls and CHC patients (P<0.05). GPC3 expression showed higher frequency in CHC patients compared to HCC group (80 vs. 28%, P=0.0016). According to the study cutoffs, serum TGF-β1 and GP73 mRNAs showed 60 and 96% sensitivities for HCC diagnosis with 100 and 95% specificities, respectively. Furthermore, elevated GP73 mRNA expression levels in early HCC were significantly increased compared to those of TGF-β1 mRNA and to high serum AFP (92.3 vs. 53.8 and 23.1%; P=0.03 and 0.0004, respectively). In conclusion, circulating TGF-β1 and GP73 mRNAs could be useful biomarkers for HCV-induced HCC diagnosis. Moreover, serum GP73 mRNA is sensitive for early cancer detection than AFP and TGF-β1 mRNA. However, these results need further validation studies.

This study aimed to evaluate the effectiveness of serum Golgi protein 73 (GP73) and Glypican-3 (GPC-3) as tumor markers for diagnosis of hepatocellular carcinoma (HCC). A total of 257 subjects were enrolled and consisted of 61 healthy controls, 32 hepatitis B virus carriers, 80 cirrhosis patients, and 84 HCC patients. Diagnosis was performed based on established clinical procedure. Serum GP73, GPC-3, and α-fetoprotein were measured. Receiving operating characteristic (ROC) curves were plotted to determine the sensitivity and specificity of each serum marker and their combinations. Serum GP73 levels were significantly increased in HCC patients. No significant differences were observed between GP73 and α-fetoprotein (AFP) as markers for HCC diagnosis. However, GP73 was more sensitive than AFP in the diagnosis of small HCC. A combination of GP73 and AFP tests increased the sensitivity and specificity for HCC diagnosis. The area under the ROC curve (AUC) of combined test was 0.93 compared with 0.88 for GP73 and 0.90 for AFP alone. GPC-3 tests were negative in all 84 HCC patients. The AUC for GPC-3 is 0.43, indicating that serum GPC-3 was not an effective tumor marker for HCC diagnosis. Serum GP73 is a potential tumor marker for HCC diagnosis, especially for differential diagnosis of small HCC and cirrhosis. The combination of GP73 and AFP is more sensitive than AFP alone. Serum GPC-3 does not appear to be an effective tumor marker for HCC diagnosis.

Early detection of hepatocellular carcinoma is very important in the treatment which is feasible. Alpha-fetoprotein plus ultrasound in surveillance programs is controversial. GP73 is a protein. Golgi increase significantly in the sera of patients with hepatitis B virus and HCV-related HCC, providing a marker for its early detection. The aim is to detect serum Golgi protein 73 (GP73) in patients with cirrhosis and with hepatocellular carci-noma (HCC), and to determine its sensitivity and specificity as a screening tool for the detection of HCC. A case control study was conducted in four groups of 32 participants each: 1- healthy controls; 2 - chronic liver disease; 3 - decompensated liver disease; 4 - HCC group. The HCC group included 25 males and 7 females with a mean age of 58 ± 7 years, fulfilling diagnostic criteria for HCC. GP73 was estimated in the serum samples taken from the HCC group and control groups. GP73 was elevated in patients with HCC and liver cirrhosis. Serum level was very high in HCC patients (p < 0.01) when compared with the other studied groups. GP73 had a sensitivity of 96.9% and specificity of 96.9% at a cutoff value of 17.5 ng/mL when compared with α-fetoprotein (AFP) that showed a sensitivity of 75% and specificity of 92% at a cutoff value of 9.4 ng/mL. GP73 can be used as a screening tool for the detection of HCC with higher diagnostic performance than AFP.

Background: Hepatocellular Carcinoma (HCC) is one of the most common malignancy in the liver. Modalities of diagnostic are often an obstacle in HCC surveillance. Alpha fetoprotein (AFP) is one of protein that often used in the diagnostic of HCC in chronic liver disease. Golgi protein 73 (GP73), is one of the candidate biomarkers in early diagnostic of HCC and found in biliary epithelial cells but rarely expressed by normal hepatocytes. Expression of GP73 was reported to be increased in a large number of malignancies. Aims of this study to evaluate differences in Golgi protein 73 serum (sGP73) and AFP in diagnosing hepatocellular carcinoma in patients with liver cirrhosis. Materials and Methods: This cross-sectional study was conducted at Haji Adam Malik Hospital in 2020. Serum level of GP73 and others biomarker was detected using enzyme-like immunosorbent assay. Results: From 90 subjects, Liver cirrhosis and HCC group had significantly higher AFP than the control group. AFP was superior in determining HCC to GP73. At a cut off value of &gt; 394.5.00 ng/mL, AFP yielded a sensitivity of 83.3% and specificity of 67%, for discriminating liver cirrhosis and HCC (AUC 0.84), while GP73 with cut off value of &gt; 82.5 ng/mL, sensitivity of 70% and specificity of 57% (AUC 0.74). Conclusion:GP73 was significantly higher in HCC patients in comparison to non-HCC patients and healthy population. Compared with alpha fetoprotein, GP73 was superior in discriminating HCC in healthy population but inferior in group of liver cirrhosis. Keywords: Golgi Protein 73, Alpha Fetoprotein, Hepatocellular carcinoma.