|Year : 2018 | Volume
| Issue : 1 | Page : 36-41
Serum level of Golgi protein-73 as a diagnostic marker for hepatocellular carcinoma
Fawzy M Khalil1, Sherif I Negm1, Mohamed A El Assal1, Anas A Yousif2, Raafat S Salem1
1 Department of Internal Medicine, Faculty of Medicine, Benha University, Banha, Egypt
2 Department of Clinical Pathology, Faculty of Medicine, Benha University, Banha, Egypt
|Date of Submission||27-Aug-2017|
|Date of Acceptance||10-Oct-2017|
|Date of Web Publication||28-Feb-2018|
Raafat S Salem
Benha Faculty of Medicine, Benha, 13511
Source of Support: None, Conflict of Interest: None
Background Golgi protein-73 (GP73) as a potential serum marker for hepatocellular carcinoma (HCC) has not been validated in studies. The aim of this study was to determine the value of serum Golgi protein-73 (SGP73) in the diagnosis of HCC.
Patients and methods SGP73 and α-fetoprotein (AFP) were compared in a total of 80 human participants in this study; 35 patients with HCC, 35 patients with cirrhosis, and 10 healthy controls were included.
Results Using 10.3 relative units as a cut-off value, the sensitivity and specificity of SGP73 for HCC were 97.1 and 85.7% compared with 57 and 55.6% for AFP (P<0.001) using 100 ng/ml as a cut-off value. The GP73 level was significantly increased in patients with HCC compared with the healthy controls (35.55 vs. 4.13%, P<0.001).
Conclusion GP73 is an accurate serum marker for the detection of HCC with higher sensitivity and specificity than AFP.
Keywords: α-fetoprotein, hepatocellular carcinoma, liver cirrhosis serum Golgi protein-73
|How to cite this article:|
Khalil FM, Negm SI, El Assal MA, Yousif AA, Salem RS. Serum level of Golgi protein-73 as a diagnostic marker for hepatocellular carcinoma. Benha Med J 2018;35:36-41
|How to cite this URL:|
Khalil FM, Negm SI, El Assal MA, Yousif AA, Salem RS. Serum level of Golgi protein-73 as a diagnostic marker for hepatocellular carcinoma. Benha Med J [serial online] 2018 [cited 2018 Sep 22];35:36-41. Available from: http://www.bmfj.eg.net/text.asp?2018/35/1/36/226411
| Introduction|| |
Hepatocellular carcinoma (HCC) is associated strongly with either chronic hepatitis B virus (HBV) or hepatitis C virus (HCV) infection ,. HCC is the fifth most common cancer and the third leading cause of cancer death worldwide. The high mortality associated with HCC is partly because of unresponsiveness to treatment, with a 5-year survival rate of less than 5% after diagnosis. In fact, most HCCs are diagnosed at an advanced stage ,. When diagnosed at an early stage, the treatment of HCC with surgical resection or liver transplantation can be curative . The prognosis can be improved significantly by early diagnosis and treatment. The use of serological markers in patients at the highest risk for developing HCC can thus decrease the cancer-related mortality and reduce medical costs . α-Fetoprotein (AFP) has been the only standard serum marker for the detection of HCC for the last 40 years, even though its sensitivity of 39–65% is not very satisfactory .
Golgi protein-73 [GP73, also called Golgi phosphoprotein-2 (GOLPH2)] is a type II Golgi-specific membrane protein that is normally expressed in epithelial cells of various human tissue types, but not hepatocytes .
However, in patients with hepatic diseases, the hepatic expression of GP73 is significantly upregulated. Therefore, GP73 might provide a solution for the early diagnosis of hepatic diseases . Subsequent studies showed that the GP73 serum level is elevated in many viral and nonviral liver diseases, including hepatitis, cirrhosis, and HCC, and also in nonliver malignances . It is significant that serum Golgi protein-73 (GP73) is markedly elevated in patients with HCC, and the sensitivity and specificity of GP73 for HCC might be superior to those of AFP .
A study by Marrero et al.  has shown that GP73 had a 62% sensitivity for the early diagnosis of hepatic carcinoma compared with 25% for AFP. Several reports have indicated that GP73 levels have an excellent clinical value for the diagnosis of PHC  and that the combined use of GP73 and AFP could even improve the detection rate of primary hepatic carcinoma (PHC) . However, Ozkan et al.  suggested that GP73 has a low value for the diagnosis and prognosis of early PHC, whereas AFP was better. However, Ozkan et al.  included patients with HCV-associated PHC and their results are still controversial .
| Aim|| |
The aim of this study was to evaluate the diagnostic value of SGP73 level in patients with HCC.
| Patients and methods|| |
This study was carried out in Faculty of Medicine, Benha University Hospitals, after obtaining approval from the Ethics Research Committee. A total of 80 individuals were enrolled; there were healthy controls (n=10), patients with cirrhosis (n=35) including HBV-positive patients (n=8), HCV-positive patients (n=23), and patients with cirrhosis of other origin (n=4), and patients with HCC (n=35). Written informed consent was obtained from the participants. The diagnosis of cirrhosis was made on the basis of clinical, laboratory, and imaging data. The symptoms and signs can include ascites, anorexia, portal hypertension, hypersplenism, and encephalopathy. The diagnosis of HCC was made on the basis of the American Association for the Study of Liver Disease (AASLD) practice guidelines.
Blood sampling and clinical chemistry
All patients underwent the standard hematological and biochemical workup for liver and kidney functions. AFP, anti-HCV antibodies, and hepatitis B virus surface antigen (HBsAg) pelviabdominal ultrasound triphasic computed tomography were performed.
Detection of serum Golgi protein-73 levels by an enzyme-linked immunosorbent assay
The test was performed using the Human GP73 enzyme-linked immunosorbent assay (ELISA) kit provided by SunRed Biotechnology Company (Shanghai, China), according to the recommendation of the manufacturer. The kit uses a double-antibody sandwich ELISA to assay the level of GP73 samples.
The collected data were tabulated and analyzed using the computer program statistical package for social science version 20 (SPSS; SPSS Inc., Chicago, Illinois, USA). Continuous variables were summarized and analyzed statistically including mean and SD, P value, an independent sample t-test, receiver operating characteristic (ROC) curve analysis, and others. P values less than 0.05 were considered to be significant and P values less than 0.005 were considered to be highly significant.
| Results|| |
The patients’ characteristics are summarized in [Table 1].
Serum Golgi protein-73 is significantly increased in patients with hepatocellular carcinoma
There were 10 healthy individuals, comprising eight women and two men with an average age of 25.2±11.5 years. As shown in [Figure 1] and [Table 2], the median GP73 value in the healthy participants was 4.13±1.43 relative units (RU). In contrast, the median value of GP73 in 35 patients with HCC was 35.55±29.68 RU), which is significantly higher than that of the healthy participants and patients with cirrhosis (8.28±4.92 RU) (P<0.001). The GP73 value of the healthy participants showed no correlation with age and sex.
|Table 2 Median α-fetoprotein and Golgi protein-73 values in different groups|
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Golgi protein-73 has higher sensitivity and specificity than α-fetoprotein in the diagnosis of hepatocellular carcinoma
As shown in [Figure 2], the area under the receiver operating characteristic (AUROC) for GP73 was 0.96 (95% CI: 0.93–1.0), with a sensitivity of 97.1%, a specificity of 85%, and an optimal cut-off value of 10.3 RU. As shown in [Figure 3], the AUROC for AFP was 0.78 (95% CI: 0.67–0.89), with a sensitivity of 57%, a specificity of 55.6%, and a cut-off of 100 ng/ml.
|Figure 2 Receiver operating characteristic curve between hepatocellular carcinoma and non-hepatocellular carcinoma in terms of Golgi protein-73.|
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|Figure 3 Receiver operating characteristic curve between hepatocellular carcinoma and non-hepatocellular carcinoma in terms of α-fetoprotein.|
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GP73 had a better AUROC compared with AFP (P<0.001), indicating higher sensitivity and specificity of GP73 compared with AFP in the diagnosis of HCC. Combined measurement of GP73 and AFP increased the sensitivity for HCC to 77.1%, with a specificity of 91.1%, and the AUROC was 0.96 (95% CI: 0.94–0.97) ([Figure 4]).
|Figure 4 Receiver operating characteristic curve between hepatocellular carcinoma and non-hepatocellular carcinoma in terms of Golgi protein-73 and α-fetoprotein.|
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The positive and negative predictive values of GP73 and AFP in the diagnosis of HCC are 85 and 97.5%, and 58.3 and 78.1%, respectively. The mean value of SGP73 in the patients with AFP levels less than 100 (cut-off value) was 21.31±6.06. The mean value of SGP73 in the patients with AFP levels higher than 100 (cut-off value) was 47.55±36.07 and this difference was significant (P<0.005) ([Table 3]).
|Table 3 Serum Golgi protein-73 levels in hepatocellular carcinoma patients with negative α-fetoprotein (<100)|
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Variations in Golgi protein-73 values in patients with different etiologies
There were no correlations between SGP73 and different etiologies in liver cirrhosis. There was significance between SGP73 and different etiologies in HCC, with a greater increase in HCV and HBV HCC patients ([Table 4]).
|Table 4 Variations in Golgi protein-73 values in patients with different etiologies|
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Golgi protein-73 levels and hepatocellular carcinoma tumor characteristics
The relationships between GP73 levels and tumor size or differentiation status or liver function were investigated. For comparison, AFP values of different tumor sizes or differentiation status or liver function were also analyzed.
GP73 values were correlated with tumor size and the Barcelona clinic liver cancer (BCLC) staging system. There was no correlation between the levels of GP73 and the number and site of focal lesion ([Table 5]).
|Table 5 Correlations between Golgi protein-73 and different classifications of the hepatocellular carcinoma group|
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The AFP value of patients with small HCCs (<3 cm) was significantly lower than that of other HCCs (>5 cm, >3 and <5 cm; P<0.001), but there was no correlation between AFP levels and BCLC staging ([Table 6]).
|Table 6 Correlations between α-fetoprotein and different classifications of the hepatocellular carcinoma group|
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| Discussion|| |
As HCC is among the cancers with the worst prognosis, early diagnosis and treatment are important for effective treatment of patients with HCC. The use of serological markers in patients at the highest risk for developing HCC may thus decrease HCC mortality and reduce medical costs. However, AFP has only been the standard serum marker for the detection of HCC for more than 40 years, even though its sensitivity of 39–65% is not satisfactory ,.
Although studies have identified SGP73 as a potential biomarker for HCC, the small sample sizes in those studies precluded the establishment of this putative tumor marker as a standard clinical test for HCC. Furthermore, whether GP73 is a better serum biomarker than AFP remains controversial and its significance in the assessment of tumor recurrence after HCC surgery remains unknown ,.
GP73 is believed to be a new serum marker for liver disease. It is a 400 amino acid 73 kDa transmembrane glycoprotein that normally resides within the cis-Golgi complex. Its mRNA was first identified in a search for upregulated hepatic genes in a patient with syncytial giant cell hepatitis .
It appears that GP73 expression is activated in hepatocytes in response to microenvironmental changes, such as necroinflammation and fibrosis. If the diseased microenvironment was improved or returned to a healthy state, hepatic GP73 expression and serum level would also return to a normal range. Multivariate regression analysis identified the SGP73 as the only independent factor for predicting hepatic inflammation and fibrosis. Thus, GP73 expression could be used as a prognostic marker for chronic liver diseases .
This study aimed to evaluate the diagnostic value of SGP73 as a tumor marker in patients with HCC.
In terms of the sero prevalence of HBsAg positivity among HCC cases, 20% of cases were positive; this was in agreement with the results obtained by Mohmad et al. , who reported that 17.5% of HCC cases were HBsAg positive. In this study, in terms of the sero prevalence of HCV Ab positivity among HCC cases, 74.3% of cases were positive. These results are slightly lower than those obtained by El-Serag , who reported that up to 90% of the HCC cases in Egypt were attributable to HCV infection. This was explained by the fact that the rate of HCV in Egypt was the highest in the world.
This study showed a statistically very highly significant increase (P<0.001) in the mean SGP73 level in the HCC group (35.5 ng/ml) compared with the cirrhotic group (8.28 ng/ml) and the control group (4.13 ng/ml). This was in agreement with Hu et al. , Tian et al. , and Mariam et al.  who reported that the serum level of GP73 is highly increased in HCC patients in comparison with either patients with benign liver diseases or the healthy controls, and has a better diagnostic performance than AFP for the detection of HCC, but not in agreement with Ozkan et al. , who reported that GP73 has a low value for the diagnosis and prognosis of early PHC whereas AFP was better.
This study also showed that there was no correlation between the levels of GP73 and the etiology of liver cirrhosis and Child’s classification. This result is in agreement with Schwegler et al. , who reported that an increase in GP73 expression is a common feature of hepatocyte response to a variety of disease etiologies.
However, the levels of GP73 showed significant differences in the size of hepatic focal lesions in the HCC group and the BCLC staging system. This finding is in agreement with Hu et al.  and Gao et al. .
This significant discrepancy in the specificity of GP73 in the diagnosis of HCC can be explained partly by differences in the disease stages of patients, but may also be a result of the different antibodies used for the ELISA in each study. In addition, the GP73 protein may consist of several subtypes, some of which may show higher HCC detection specificity and specificity .
How GOLPH2 functions and the mechanisms of regulation in normal and neoplastic tissues are still unclear. Possibly, it is either involved in post-translational protein modification, transport of secretory proteins, cell signaling regulation, or simply maintenance of Golgi apparatus function. GOLPH2 has several potential glycosylation sites and up to 75% of GOLPH2 secreted from hepatocytes are fucosylated .Once hepatic necrosis was triggered, the affected hepatocytes released more GP73 into the blood, resulting in elevated SGP73 concentrations. In the meantime, an inflammatory reaction, including an influx of infiltrating lymphocytes, occurred at the necrotic sites. The infiltrated immune cells may produce and release higher concentrations of proinflammatory cytokines in the liver. In response to this increased inflammatory condition, or in response to other pathologic changes such as fibrosis and carcinogenesis, hepatocytes may show an increase in the expression of GP73. This would explain why hepatic GP73 expression and SGP73 levels were significantly increased in patients with severe hepatitis, cirrhosis, and HCC .
In addition to liver injury-mediated GP73 release from hepatocytes, other mechanisms, including induction and regulation by interleukin-6 or interferon-g, may facilitate GP73 release .
It appears that GP73 expression is activated in hepatocytes in response to microenvironmental changes, such as necroinflammation and fibrosis. If the diseased microenvironment was improved or returned to a healthy state, hepatic GP73 expression and serum levels would also return to a normal range .
| Conclusion|| |
The data obtained in our study indicate that GP73 is a valuable serum marker that can aid the diagnosis of HCC. In combination, measurements of AFP and GP73 have the potential to further improve the detection and treatment of one of the most common malignancies worldwide.
The authors thank Dr Ahmed Mohammed Hussein Dabour for his support and fruitful efforts to complete this work.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
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[Figure 1], [Figure 2], [Figure 3], [Figure 4]
[Table 1], [Table 2], [Table 3], [Table 4], [Table 5], [Table 6]