Joint Detection of GP73 and AFP-L3 in Primary Hepatic Carcinoma with Low Concentration of AFP J Int Transl Med, 2015, 3(1):28-32; doi: 10.11910/2227-6394.2015.03.01.06 Open Access
Research Article
Diagnostic value of Joint Detection of GP73 and AFP-L3 in Primary Hepatic Carcinoma with Low Concentration of AFP CAI Lei1, RAO Xiao-hui2, SU Qi3, QIN Jia-sheng1, CAI Li-quan1, HONG He1, WANG Kang-hua1, AI Zhi-guo2, ZHANG Sheng2 1. Second Department of Hepatobiliary Surgery, Zhujiang Hospital, State Key Laboratory of Organ Failure Research, Co-Innovation Center for Organ Failure Research, Southern Medical University, Guangzhou, 510280, China; 2. Department of Hepatobiliary Surgery, Huizhou Municipal Central Hospital, Huizhou, 516001, China 3. Radiology Department, Qinhuangdao Chinese Medical Hospital, Qinhuangdao, 066002, China.
ABSTRACT Objective: To explore the applicative value of serum Golgi protein (GP73) and alpha-proteinvariant (AFP-L3) in the diagnosis of patients with primary hepatic carcinoma (PHC). Methods: Totally 110 patients were enrolled, including 60 PHC patients with low concentration of AFP ((1.1 ～ 108.0 μg/L)) and 50 patients with non-PHC digestive system diseases (20 cases of patients with chronic hepatitis, 15 patients with liver cirrhosis, 4 patients with bile duct cancer, 4 patients with gastric cancer, 4 patients with rectal cancer and 3 patients with colon cancer). In addition, 42 healthy people were selected as control group. GP73 was detected by enzyme-linked immunosorbent assay (ELISA) trace centrifugal column method was adopted for separation of AFP-L3. Luo’s chemiluminescence method was used to determine the total content of AFP and AFP-L3 in eluent for calculating the ratio of AFP-L3/ AFP. Results: The levels of serum GP73 and AFP-L3(%) in PHC group were significantly higher than the other 2 groups (P ＜ 0.01) and the level of serum GP73 and AFP-L3 (%) in Non-PHC group were higher than those in healthy group (P ＜ 0.01). ROC analysis showed that the area under ROC curve of single GP73 and AFP-L3(%) in diagnosis of PHC and non-PHC were 0.887 and 0.860, respectively. Additionally, the ROC analysis also showed that critical value of GP73 and AFP-L3 for the diagnosis of HPC were 83.78 μg/L and 13.87%, respectively. The sensitivity of joint detection of serum GP73 or AFP-L3 was higher than detection of them alone (90.0 vs. 71.7 and 60.0, P ＜ 0.05) but the specificity was similar between single detection and joint detection (P ＞ 0.05). The positive predictive value and negative predictive value of joint detection of them was higher than single detections of them and there was significant differences in negative predictive
Key words:
value (P ＜ 0.05) but no difference in positive predictive value (P ＞ 0.05). The overall response
Primary hepatic carcinoma
rate of joint detection of GP73 and AFP-L3 was higher than single detection of them (P ＜ 0.05).
Golgi glycoprotein 73
Conclusion: single detection of GP73 and AFP-L3 (%) were similar in the sensitivity and
Alpha-fetoprotein-L3
specificity in diagnosis of PHC patients with low concentration of AFP, which can be auxiliary
Diagnostic value
diagnosis for PHC patients and joint detection of them is more reliable to diagnose PHC.
28
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J Int Transl Med, 2015, 3(1):28-32 in average (range 35 ～ 79 years). All patients conformed to viral
Introduction Primary hepatic carcinoma (PHC) is one of malignant tumors with the highest morbidity rate in the world and has a trend [1]
of rising year by year at present . It is the third leading cause of cancer deaths worldwide [2] and the third of China in the [3]
morbidity of malignant tumors . The 5-year survival rate of patients with liver cancer is no more than 10%. The early diagnosis is the main effective strategy of treating PHC and prolonging survival time of PHC patients. Although alphafetoprotein (AFP) is the most commonly-used tumor marker for examining PHC, its sensitivity and specificity are not satisfactory. Moreover, AFP is not accurate for examining low concentration of AFP in patients with liver cancer[4]. Therefore, seeking for tumor markers with higher sensitivity and specificity and diagnostic methods are of significance to clinical treatment of HPC. Serum glogi glycoprotein 73 (GP73) and alphafetoprotein-L3 (AFP-L3) is expected to become mew tumor [5-6]
markers for the early detection and prognosis of PHC In 2005, Block et al
[7]
.
proposed that GP73 level increased
significantly in the serum of patients with liver cancer. In the same year, Marrero et al
[8]
found that the sensitivity and
specificity of GP73 in diagnosis of hepatocellular carcinoma (HCC) was more than 70% and the sensitivity of GP73 were 2.55 fold of sensitivity of AFP in diagnosis of early HCC. When AFP is less than 20 ng/L, GP73 level is increasing obviously in over half of patients with HCC. So for PHC patients with negative AFP, application of GP73 can obviously enhance the detection rate of PHC[9-10]. Another study have proposed that the combination of GP73 and other tumor markers can improve the diagnostic significance of PHC, but the application of joint detection is still to be verified by clinical data. Hence, this study used enzyme-linked immunosorbent assay (ELISA) to detect the levels of serum GP73 and AFP-L3 and evaluate the efficiency of joint detection of GP73 and AFP-L3, so as to improve the clinical application value of diagnosis and early warning of early PHC.
hepatitis diagnostic criteria of Viral Hepatitis Prevention Scheme revised by Chinese Medical Association Infectious Diseases and Parasitic Epidemiology Branch Liver Disease (2001) and PHC diagnostic criteria of Chinese Anti-Cancer Association. The other 50 cases were patients with non-PHC digestive system diseases, 31 males, 19 females, aged 54 years (range 36 ～ 81 years), including 20 cases of patients with chronic hepatitis (Serum hepatitis b or hepatitis c virus detection showed positive) in whom liver cirrhosis and liver lesions were ruled out in accordance with clinical altrosound and CT examinations, 15 patients with liver cirrhosis in whom liver lesions were ruled out, and 4 patients with bile duct cancer, 4 patients with gastric cancer, 4 patients with rectal cancer and 3 patients with colon cancer. In addition, 42 healthy people were selected as control group, in whom there were 20 males and 22 females, aged 55 years (range 36 ～ 82 years). Detection of serum hepatitis virus, ALT showed negative and hepatitis, liver cirrhosis, liver and bile cyst and benign and malignant tumors were ruled out. All the patients agreed to participate in this study and signed the informed consent form.
Key reagents and instruments CODA automatic enzyme standard instrument and electrochemical luminescence instrument were bought from Hamilton Bonaduz AG，Switzerland. GP73 kit was purchased from Roche Diagnostics (Shanghai) Ltd. AFP-L3 separation and purification kit were purchased from Beijing Hot Scene Biological Technology Co., Ltd.
Collection of specimen The fasting blood (3 mL) was taken from cubital vein by procoagulation tube in the early morning, centrifuged at 3 000 r/min for 10 min, with the centrifugal radius being 16 cm. The serum was separated and preserved at -80℃ .
Detection method According to the specification, GP73 was detected by enzyme-
Materials and Methods
linked immunosorbent assay (ELISA) and trace centrifugal
Study objects
chemiluminescence method was used to determine the total
column method was adopted for separation of AFP-L3. Luo’s
A total of 110 patients at Outpatient Department of Second
content of AFP and AFP-L3 in eluent for calculating the ratio of
Department of Hepatobiliary Surgery, Zhujiang Hospital were
AFP-L3/ AFP.
selected, among which, there were 60 PHC patients with low concentration of AFP (1.1 ～ 108.0 μg/L) detected by
Statistical analysis
electrochemiluminescence, 39 males, 21 females, aged 53 years
SPSS13.0 software package was used for data analysis. The
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J Int Transl Med, 2015, 3(1):28-32 detecting data were in skewed distribution and expressed by ( x ± s ). Comparison among multiple groups was analyzed using t test. Different groups of measurement data was used analysis of variance. Comparison of enumeration data and ratio were analyzed using X2 test. Inspection level is α ＝ 0.05, and P ＜ 0.05 was considered to be significantly significant.
Results Detection results of serum GP73 and AFP-L3 in different groups The levels of serum GP73 and AFP-L3(%) in PHC group were significantly higher than the other 2 groups (P ＜ 0.01) and the level of serum GP73 and AFP-L3 (%) in Non-PHC group were higher than those in Healthy group (P ＜ 0.01), as shown in Table 1.
was higher than detection of them alone (P ＜ 0.05) but the specificity was similar between single detection and joint
Table 1 Comparison of Serum GP73 and AFP-L3 Levels in Different Groups ( x ± s ) Groups
Figure 1 ROC Curve Analysis of GP73 and AFP-L3 in the Diagnosis of PHC.
detection (P ＞ 0.05). The positive predictive value and negative predictive value of joint detection of them was higher than single detections of them and there were significant differences
n
GP73 (μg/L)
AFP-L3 (%)
PHC group
60
185.65±52.84**
15.40±4.25**
positive predictive value (P ＞ 0.05). The overall response rate
Non-PHC group
50
60.22±23.46 △△
9.45±4.66 △△
of joint detection of GP73 and AFP-L3 was higher than single
Healthy group
42
41.76±19.30
6.64±3.84
detection of them (P ＜ 0.05). However, single detections of
Compared with Healthy and Non-PHC groups, **P ＜ 0.01. Compared with Healthy group, △△ P ＜ 0.01. Notes: PHC group refers to PHC patients with low concentration of AFP (1.1 ～ 108.0 μg/L); Non-PHC group refers to other digestive system diseases except from PHC patients of low concentration of AFP.
ROC curve analysis of single detection of serum GP73 or AFP-L3 in differential diagnosis of PHC ROC analysis showed that the area under ROC curve of single GP73 and AFP-L3(%) in diagnosis of PHC and non-PHC were 0.887 (confidence interval 0.823 ～ 0.951, P ＝ 0.000) and 0.860 (0.790 ～ 0.930, P ＝ 0.000), with the standard error being 0.033
in negative predictive value (P ＜ 0.05) but no difference in
them were similar in all testing indexes. (Table 2) Table 2 Diagnostic Value of Single Detection of Serum GP73 or AFP-L3(%) and Joint Detection for HPC GP73
AFP-L3
GP73+ AFP-L3
Sensitivity
71.7(43/60)
60.0(36/60)
90.0(54/60)
Specificity
88.0(81/92)
92.4(85/92)
93.5(86/92)
Positive predictive value
86.0(43/50)
80.0(36/45)
94.8(55/58)
Negative predictive value
79.4(81/102)
79.4(85/107)
91.5(86/94)
Overall response rate 81.6(124/152) 79.6(121/152) 92.1(140/152)
and 0.036, respectively. Additionally, the ROC analysis also
Discussion
showed that critical value of GP73 and AFP-L3 for the diagnosis
Early detection of PHC is one of the hot issues for diagnosis and
of HPC were 83.78 μg/L and 13.87%, respectively. (Figure 1)
treatment of liver cancer. At present, although AFP is the most
Diagnostic value of single detection of serum GP73 or AFP-L3 (%) and joint detection for HPC The sensitivity of joint detection of serum GP73 or AFP-L3
30
commonly-used tumor marker of serum diagnostic markers for PHC and is widely applied in the screening, clinical diagnosis, evaluation of efficacy and prediction of reoccurrence of PHC, its sensitivity and specificity are only up to 60% ～ 70%, which is not satisfactory. For another reason, AFP shows false negative in
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J Int Transl Med, 2015, 3(1):28-32 early liver cancer, especially in small hepatocellular carcinoma,
0.887and 0.860, respectively, with the standard error being 0.033
and serum AFP increase when liver cells are in the process of
and 0.036, respectively. Additionally, the ROC analysis also
damage and regeneration. In recent years, researchers at home
showed that critical value of GP73 and AFP-L3 for the diagnosis
and abroad are trying to seek for new tumor markers of more
of HPC were 83.78 μg/L and 13.87%, respectively. Moreover,
sensitivity and specificity for the diagnosis of PHC. AFP-L3 and
the sensitivity of joint detection of serum GP73 or AFP-L3
GP73 were new serum markers for diagnosis of PHC[11].
was higher than detection of them alone but the specificity was similar between single detection and joint detection. The
AFP-L3 is produced by liver cancer cells, revealing the tendency
positive predictive value and negative predictive value of joint
of vascular invasion and metastasis of liver cancer. The higher
detection of them was higher than single detections of them and
the percentage of AFP-L3/AFP, the malignant degree the tumors
there was significant difference in negative predictive value but
are. AFP-L3 is not expressed in cells of benign liver diseases, so
no difference in positive predictive value. The overall response
AFP-L3 of high specificity is conductive to the early diagnosis,
rate of joint detection of GP73 and AFP-L3 was higher than
differential diagnosis, efficacy evaluation and prognosis of
single detection of them. However, single detections of them
PHC[12-13]. GP73 is a kind of type Ⅱ transmembrane protein and
was similar in all testing indexes. Therefore, we held that single
is expressed by biliary epithelial cells in human body but less
detection of serum GP73 had little disadvantages for diagnosis
expressed or no expressed in normal liver cells. However, it is
of PHC but joint detection of GP73 and AFP-L3 could improve
[14]
pretty active in PHC . That is because GP73 arrives endosome
the diagnostic rate of PHC, especially in achieving early
and cell surface out from Golgi apparatus cis Golgi, thus leading
discovery, detection and treatment in PHC patients without
[15]
to the abnormal increase of serum GP73 in patients with PHC .
clinical symptom, signs and negative AFP, thus prolonging the
Additionally, the increased level of GP73 is not only related to
survival time of the patients.
patients infected with hepatitis b virus but also closely associated with liver cell damage in patients with PHC or the function of [16]
In conclusion, single detection of GP73 and AFP-L3 (%)
.
was similar in the sensitivity and specificity in diagnosis of
Relevant studies have reported that the level of serum GP73 in
PHC patients with low concentration of AFP, which can be an
patients with PHC is higher than that in patients with benign
auxiliary diagnosis for PHC patients and joint detection of them
lever cancer and healthy people, showing that GP73 is more
is more reliable to diagnose PHC. Samples in the study are small
GP73 keeping the structural integrity of Golgi apparatus
effective liver cancer markers than AFP
[17-18]
. Additionally,
multiple researches are found that the expression of GP3 is not associated with tumor size, age of study objects, and though
so the accurate outcomes are still to be further study in large clinical evidence.
it increases in patients with hepatitis and liver cirrhosis but it
Declaration
is more obviously in PHC, which proves the value of GP73 in
All authors of this article declare no conflict of interest.
diagnosis of PHC[19-20].
Acknowledgement
In this study, the levels of CP73 and AFP-L3 (%) of 60 PHC
The study was approved by Second Department of Hepatobiliary
patients with low concentration of AFP were detected for
Surgery, Zhujiang Hospital, State Key Laboratory of Organ
evaluation of the detecting efficacy of GP73 and AFP-L3 (%)
Failure Research, Co-Innovation Center for Organ Failure
on PHC with low concentration of AFP. And the results showed
Research, Southern Medical University, Guangzhou, China
that the levels of serum GP73 and AFP-L3(%) in PHC patients
and co-funded by the National High Technology Research
with low concentration of AFP were significantly higher than
and Development Program of China (“863”Program
non-PHC and healthy people and the level of serum GP73 and
2012AA020505), the National Natural Science Foundation of
AFP-L3 (%) in non-PHC patients were higher than those in
China (81470875).
healthy people (P ＜ 0.01), showing that the content of serum GP73 and AFP-L3 was closely related to the liver damage and
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