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Diagnostic accuracy of PIVKA-II, ALPHA-Fetoprotein and a combination of both in diagnosis of Hepatocellular Carcinoma

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Patients affected by liver cirrhosis are at high risk for developing hepatocellular carcinoma. The aim of this study was to evaluate the feasibility of PIVKA-II (protein induced by vitamin K absence or antagonist-II) alone or in combination with α-1 fetoprotein, as a screening marker for development of hepatocellular carcinoma. Subjects and methods: A case-control study was conducted on 103 Military Hospital. 53 patients affected by hepatocellular carcinoma were considered as cases, while 31 patients affected by liver cirrhosis were considered as control. Results: At the set cut-off value of 40 mAU/mL, PIVKA-II was more sensitive (92.45% vs. 81.13%), but less specific than α-1 fetoprotein at the set cut-off value of 10 ng/mL (61.29% vs. 83.87%). The negative predictive value of PIVKA in combination with α-1 fetoprotein was 88.23% and the sensitive was 96.2%. Conclusion: PIVKA- II was more sensitive but less specific than α-1 fetoprotein in diagnosis of hepatocellular carcinoma and PIVKA-II, when combined with α-1 fetoprotein, may be considered as a screening test for hepatocellular carcinoma due to its high sensitive and negative predictive value.

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DIAGNOSTIC ACCURACY OF PIVKA-II, ALPHA-FETOPROTEIN

AND A COMBINATION OF BOTH IN DIAGNOSIS OF

HEPATOCELLULAR CARCINOMA

Pham Van Tran 1 ; Hoang Thi Minh 1

SUMMARY

Background/aim: Patients affected by liver cirrhosis are at high risk for developing hepatocellular carcinoma The aim of this study was to evaluate the feasibility of PIVKA-II (protein induced by vitamin K absence or antagonist-II) alone or in combination with α-1 fetoprotein, as a screening marker for development of hepatocellular carcinoma Subjects and methods: A case-control study was conducted on 103 Military Hospital 53 patients affected by hepatocellular carcinoma were considered as cases, while 31 patients affected by liver cirrhosis were considered as control Results: At the set cut-off value of 40 mAU/mL, PIVKA-II was more sensitive (92.45%

vs 81.13%), but less specific than α-1 fetoprotein at the set cut-off value of 10 ng/mL (61.29%

vs 83.87%) The negative predictive value of PIVKA in combination with α-1 fetoprotein was 88.23% and the sensitive was 96.2% Conclusion: PIVKA- II was more sensitive but less specific than α-1 fetoprotein in diagnosis of hepatocellular carcinoma and PIVKA-II, when combined with α-1 fetoprotein, may be considered as a screening test for hepatocellular carcinoma due to its high sensitive and negative predictive value

* Keywords: Cirrhosis; Hepatocellular carcinoma; PIVKA-II; α-1 fetoprotein

INTRODUCTION

Chronic hepatitis, cirrhosis, hepatocellular

carcinoma (HCC) are common and

dangerous diseases of the liver Currently,

there are many supportive methods for

the diagnosis, treatment monitoring and

prognosis of these diseases Non-invasive

diagnostic methods have been developed

One of these methods is the quantification

of biomarkers circulating in the blood

This method is easy to use and can be

carried out regularly

The most commonly utilized biomarker

in clinical practice for HCC is α-1 fetoprotein

(AFP) AFP may be used as a diagnostic

and prognostic marker Serum AFP levels also correlate with tumor size, tumor doubling time and post-treatment relapse [1, 2] However, it is not recommended for routine screening because it lacks specificity AFP may be elevated in other malignancies

(e.g gastric, embryonic cancer, intrahepatic

cholangiocarcinoma), as well as in many

benign conditions, both hepatic (e.g cirrhosis,

necrosis, acute hepatitis), and extrahepatic

(e.g pneumonia) Finally, AFP lacks sensitivity;

serum AFP levels are high in only 40 - 60%

of HCC cases and only 10 - 20% of early HCC [2] For these reasons, more sensitive and specific HCC markers are required

1.103 Military Hospital

Corresponding author: Pham Van Tran (phamvantran@yahoo.fr)

Date received: 20/12/2018

Date accepted: 21/01/2019

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PIVKA-II (protein induced by vitamin K

absence or antagonist II) also known as

des-gamma-carboxyprothrombin - DCP),

is an immature form of prothrombine without

any coagulative function It is synthesized

in the presence of acquired defects of

precursor carboxylation found in patients

with HCC [5] PIVKA-II may have a role in

HCC progression by acting as an autologous

growth factor [6] Moreover, high serum

levels of PIVKA-II are associated with

HCC size, microvascular invasion, metastatic

dissemination and HCC relapse after liver

transplantation and/or HCC nodule ablation

PIVKA-II is a specific marker for HCC, it is

poorly related to AFP and exhibits higher

sensitivity and specificity than AFP in

diagnosing HCC PIVKA-II is not elevated

in benign hepatic diseases (e.g cirrhosis,

necrosis), while it is elevated in HCC at

early stage as well as in HCC nodules of

all sizes For these reasons, some authors,

and especially Japanese guidelines on

HCC management proposed the use of

PIVKA-II for routine follow-up in patients

with high risk for developing HCC, as well

as a prognostic biomarker before surgery

to predict microvascular invasion, relapse

and overall poor prognosis [4, 8] In detail,

Japanese guidelines have suggested to

consider as a diagnostic marker of HCC with

high elevation of tumor markers, namely

AFP ≥ 200 ng/mL and PIVKA-II ≥ 40 mAU/mL

However, data on its performance as a

diagnostic marker are still incomplete

The aim of this study was: To evaluate

the feasibility of PIVKA-II (protein induced

by vitamin K absence or antagonist-II)

alone or in combination with AFP, as a

screening marker for development of HCC

SUBJECTS AND METHODS

A case-control study was performed in Gastroenterology Department, 103 Military

Hospital 53 patients affected by HCC were

considered as cases, while 31 patients affected by liver cirrhosis were considered

as control

All patients are diagnosed definitively and grouped based on underwent history-taking, physical examination, routine laboratory examinations and abdominal imaging either by abdominal ultrasonography, CT-scanning Cases: Cytological evidence of HCC as assessed in AASLD Hepatocellular Carcinoma Guidelines [2] Controls: Chronic hepatitis, cirrhosis and absence of nodular hepatic lesions under abdominal ultrasonography

In each patient: Take 2 mL of anticoagulant blood with heparin after fasting for at least 8 hours Centrifugal plasma separator AFP and PIVKA-II assay was performed using Architect ci16200 system of Abbott Company based on the principle of immunization with microcrystalline luminescence

Statistical analysis was performed using software package SPSS statistic 2.2

RESULTS

Table 1: Characteristics of patients (n = 84)

(n = 53)

Control

No differences were observed in gender and age between the two groups

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RESULTS

Table 1: Characteristics of patients (n = 84)

No differences were observed in gender and age between the two groups

Figure 1: The common clinical symptoms in two groups

In HCC, the common clinical symptoms were fatigue and right lower back pain

Other symptoms such as weight loss, gastrointestinal disorders were less frequent and

are comparable to patients with chronic liver disease

Table 2: Characteristics of tumor on abdominal ultrasonography and CT

Location

Number of tumor

Patients with primary tumors were on the right liver, followed by both lobes

Most patients had a tumor The results of abdominal ultrasonography and CT were not

uniform across all parameters

0 10

20

30

40

50

60

70

80

Cases Control

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Table 3: Concentration of AFP and PIVKA-II

Cases (n = 53) Median (min - max)

Controls (n = 31)

Both serum AFP (1286.28 vs 4.59 ng/mL, p < 0.001) and serum PIVKA-II concentrations (5.742 vs 27.10, p < 0.001) were higher in the case group

Figure 2: Linear correlation between AFP and PIVKA-II in HCC

Serum levels of AFP and PIVKA-II were correlated (ρ < 0.05)

Figure 3: Received operating characteristics curve (ROC) comparing specificity and

sensitivity of PIVKA-II and AFP in detecting HCC

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The performance of both PIVKA-II and AFP in the diagnosis of HCC were plotted on

a ROC curve (figure 2) showing similar areas under curve for both markers; specifically,

areas under curve (AUC) for PIVKA-II (cut-off: 40 mAU/mL) and AFP (cut-off: 10 ng/mL) were 0.864 (95%CI = 0.777 - 0.950) and 0.925 (95%CI = 0.870 - 0.981), respectively

Table 4: Sensitivity, specificity, positive predictive value (PPV) and negative predictive

value (NPV) of AFP, PIVKA-II and the combination of both in detecting HCC

> 40 mAU/mL

At the set cut-off value of 40 mAU/mL, PIVKA-II was more sensitive (92.45% vs 81.13%) but less specific than AFP at the set cut-off value of 10 ng/mL (61.29% vs

83.87%) The negative predictive value of PIVKA in combination with AFP was 88.23% and it was higher when compared both to AFP (72.22%) and PIVKA alone (82.6%)

(table 2)

DISCUSSION

HCC is the sixth most common cancer

in the world and the third most common

cause of cancer-related death In our

study, patients with HCC were older than

patients without HCC and men dominate

the two groups Male predominance may

be explained by greater exposure to

toxins, HCV prevalence and hepatocyte

androgenic stimulation [9] Imaging

methods play an important role in the

diagnosis and evaluation of treatment

outcome as well as relapse after

treatment In clinical practice today,

ultrasound and CT are the most widely

used method We conducted simultaneous

study of ultrasonographic and CT results

of patients who studied, the results were

quite similar between the two methods

with liver tumors mainly concentrated in the right liver, a few patients have tumor

in the left liver and most patients had a

tumor (table 2)

Patients with cirrhosis, especially if due

to HBV or HCV, are at high risk for developing HCC and, therefore, undergo active surveillance with six-monthly upper abdominal ultrasound However, researches for the identification of an effective serum marker for screening of HCC are currently ongoing AFP is widely used for this purpose, even though it is not recommended by neither EASL nor AASLD Guidelines because of its low sensitivity and specificity [9] AFP is elevated in only 40 - 60% of cases of HCC Sensitivity is even lower in early HCC cases, with serum elevation of AFP

in only 10 - 20% of cases Moreover,

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as a non-specific inflammation and

regeneration marker, it may also be

elevated in liver diseases (e.g liver

cirrhosis and hepatic necrosis), as well as

in some extra-hepatic conditions such as

pneumonia and embryonal tumors [3]

PIVKA-II is an abnormal prothrombine

due to carboxyl group deficiency at

gamma position in 10 glutamic acid

molecules located at the N-terminal side

of the prothrombine Carboxylation

(COOH binding) is dependent on the

presence of vitamin K The loss of

carboxyl group in 10 glutamic acid

molecules should reduce the ability of

Ca2+ to bind to prothrombine and may

cause reversibility normal blood clotting

The role of PIVKA-II in liver cancer was

first reported in 1984 by Liebman H.A et

al In the first time, the role of PIVKA-II in

the diagnosis, prognosis, evaluation of the

effectiveness of HCC treatment was

studied as well as the most clinical

application in Japan In recent times, role

of PIVKA-II has been paid more attention

to research by the United States, some

European countries, as well as in Asia In

chronic liver disease and HCC, the

biological processes of AFP and PIVKA

are independent of each other Most

studies in Japan and Asia have shown

that PIVKA-II plays a better role than

AFP, especially with respect to tumor

size, correlation with thrombosis or

invasion of blood vessels, assess the

possibility of relapse after treatment [5]

When conducting AFP and PIVKA-II levels

in patients with chronic liver disease and

HCC, performing a quantitative test on the

same principle and the same analytical system we found that both plasma PIVKA-II and AFP levels were statistically significantly different in patients with chronic hepatitis and cirrhosis This is also consistent with many other studies in the world as authored by Inagaki Y et al [5] Similarly to other authors, we observed correlation between PIVKA-II and AFP serum levels This allowed us to evaluate the combined performance of both markers in the diagnosis of HCC, as suggested by already available evidence [10] When cut-off values of 40 mAU/mL for PIVKA-II and 10 ng/mL for AFP were selected, we showed that PIVKA-II had a higher sensitivity but a lower specificity than AFP Combining both markers further increased overall sensitivity and negative predictive value at the expense

of specificity (88.23%) These results slightly differed from a recent French study in which the authors showed that PIVKA-II had a better performance than AFP for early HCC diagnosis with a

sensitivity of 77% (vs 61%), a specificity

of 82% (vs 50%), a positive predictive value of 76% (vs 51%) and a negative predictive value of 83% (vs 62%), at a

cut-off values of 42 mAU/mL for PIVKA-II and 5.5 ng/mL for AFP [7]

CONCLUSION

PIVKA-II was more sensitive but less specific than AFP in diagnosis of HCC and PIVKA-II, when combined with AFP, may be considered as a screening test for HCC due to its high sensitive and negative predictive value

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