Home About us Editorial board Search Ahead of print Current issue Archives Submit article Instructions Subscribe Contacts Login 


 
 Table of Contents  
ORIGINAL ARTICLE
Year : 2017  |  Volume : 30  |  Issue : 3  |  Page : 887-891

Plasminogen activator inhibitor-1 SERPINE1 4G/5G polymorphism in hepatocellular carcinoma patients


Department of Clinical Pathology, Faculty of Medicine, Menoufia University, Shebein El-Kom, Egypt

Date of Submission10-May-2016
Date of Acceptance26-Jun-2016
Date of Web Publication15-Nov-2017

Correspondence Address:
Sara A. M. Hegazy
Department of Clinical Pathology, Faculty of Medicine, Menoufia University, Shebein El-Kom, Menoufia Governorate, 32511
Egypt
Login to access the Email id

Source of Support: None, Conflict of Interest: None


DOI: 10.4103/1110-2098.218258

Rights and Permissions
  Abstract 

Objective
The aim of this study was to investigate the distribution of genotypes and the frequency alleles of plasminogen activator inhibitor-1 (PAI-1) 4G/5G polymorphism in patients with hepatocellular carcinoma (HCC).
Background
PAI-1 4G/5G polymorphism involves a guanosine insertion/deletion in the promoter region of SERPINE1 gene at the –675 bp position.
Patients and methods
This study was conducted on 49 HCC patients (39 male and 10 female). Their ages ranged between 44 and 84 years. They were selected from the inpatient and outpatient clinics of Shibin El-Kom Fever Hospital. Patients were genotyped for PAI-1 4G/5G polymorphism using PCR.
Results
In HCC patients, there were no statistically significant differences when comparing carriers of the three genotypes (5G/5G, 4G/5G, and 4G/4G) as regards sex, Child score, total bilirubin, direct bilirubin, albumin, serum glutamate pyruvate transaminase, alkaline phosphatase, γ-glutamyltranspeptidase, international normalized ratio, prothrombin, and α-fetoprotein (P = 0.81, 0.76, 0.46, 0.054, 0.99, 0.92, 0.22, 0.58, 0.49, 0.42, 0.54, and 0.85, respectively). On comparing the three genotypes (5G/5G, 4G/5G, and 4G/4G) among HCC patients as regards serum glutamic oxaloacetic transaminase, there was a statistically significant difference among them (P = 0.02). On comparing carriers of the three genotypes (5G/5G, 4G/5G, and 4G/4G) as regards different tumor characters (tumor size, tumor number, tumor side, and Barcelona Clinic Liver Cancer staging system), there were no significant differences among them (P = 0.16, 0.59, 0.42, and 0.14, respectively). Rate of thrombosis in the HCC group was 62.5% in 4G/5G genotype versus 41.2 and 12.5% in 5G/5G and 4G/5G genotype, respectively; there was a statistically significant difference (P = 0.04).
Conclusion
It could be concluded that PAI-1 4G/5G polymorphism was not associated with HCC.

Keywords: hepatocellular carcinogenesis, SERPINE1 4G/5G polymorphism, plasminogen activator


How to cite this article:
El-EdeL RH, Essa ES, Essa AS, Hegazy SA. Plasminogen activator inhibitor-1 SERPINE1 4G/5G polymorphism in hepatocellular carcinoma patients. Menoufia Med J 2017;30:887-91

How to cite this URL:
El-EdeL RH, Essa ES, Essa AS, Hegazy SA. Plasminogen activator inhibitor-1 SERPINE1 4G/5G polymorphism in hepatocellular carcinoma patients. Menoufia Med J [serial online] 2017 [cited 2020 Apr 8];30:887-91. Available from: http://www.mmj.eg.net/text.asp?2017/30/3/887/218258


  Introduction Top


Hepatocellular carcinoma (HCC) accounts for more than 5% of all human cancers and for 80–90% of primary liver cancer. It is a major health problem worldwide, being the fifth most common malignancy in men and the eighth in women; it is the third most common cause of cancer-related death in the world [1].

In Egypt, HCC is reported to account for about 4.7% of chronic liver disease patients [2]. The prevalence of antihepatitis C virus antibodies among Egyptian patients with HCC ranges from 70 to 80% [3].

In the multistep process of HCC development, both the chemical carcinogens and the oncogenic viruses can cause DNA damage, which is manifested at the chromosome level as deletions, duplications, and translocations, frequently affecting the structure and expression of cancer-related genes [4].

Plasminogen activator inhibitor-1 (PAI-1) is a globular protein with three β sheets (A, B, and C) and nine α-helices (hA–hI) [5]. Evidence has emerged indicating that PAI-1 can modulate angiogenesis [6] and regulate cell attachment, detachment, migration, and invasion, thus facilitating carcinogenesis [7].

The PAI-1 is encoded by the SERPINE1 gene located at 7q21.3–q22 and consists of nine exons. Several polymorphisms in the promoter region have been identified [8] – for example, a guanosine insertion/deletion polymorphism in the promoter region of SERPINE1 gene at the –675 bp position, named 4G/5G [9].


  Patients and Methods Top


This study was conducted on 49 HCC patients (39 male and 10 female) presented to Shibin El-Kom Fever Hospital during the period from March 2013 to February 2015. Patients were genotyped for PAI-1 4G/5G polymorphism using PCR. The study protocol was approved by the ethical committee in Menoufia University. Informed consent was taken from the patients before the beginning of the study.

Plasminogen activator inhibitor-1 4G/5G genotyping assay

DNA was extracted from EDTA-treated blood sample using Thermo Scientific Gene JET Genomic DNA Purification Kit (Thermo Fisher Scientific, Waltham, Massachusetts, USA). Isolated DNA was amplified using PCR.

DNA amplification for molecular detection of SERPINE1 promoter 4G/5G polymorphism was performed with an allele-specific (PCR) analysis using the following specific primers: insertion 5G allele, 5'-GTC TGGACA CGT GGG GG-3'; deletion 4G allele, 5'-GTC TGG ACA CGTGGG GA-3'; each in a separate PCR reaction together with the common downstream primer (5'-TGC AGC CAG CCA CGT GATTGT CTA G-3') and a control upstream primer (5'-AAG CTT TTACCA TGC TAA CCC CTG GT-3') to verify the occurrence of DNA amplification in the absence of the allele on the genomic DNA. PCR cycling conditions included an initial denaturation step at 95°C for 10 min, followed by 35 cycles; each cycle consisted of denaturation at 94°C for 1 min, annealing at 54°C for 30 s, extension at 72°C for 40 s, and final extension of 5 min at 72°C. PCR mix without DNA sample was used to ensure contamination-free PCR product. The amplified DNA fragments were then run on 2% agarose gel using gel electrophoresis and visualized on a ultraviolet transilluminator to detect the presence of amplified material. Each study participant was classified into one of the three possible genotypes: 4G/4G, 4G/5G, or 5G/5G according to the presence of the 139 bp PCR product generated by the allele-specific primers [Figure 1] and [Figure 2].
Figure 1: A representive agarose gel electrophoresis for 5G allele. Lane 1: marker, ladder 100 bp; lanes 2, 4 positive allele bands at 139 bp; lane 3, an negative allele.

Click here to view
Figure 2: A representive agarose gel electrophoresis for 4G allele. Lane 1: marker, ladder 100 bp; lanes 2 and 3: positive allele bands at 139 bp; lanes 4 negative allele.

Click here to view


Statistical analysis

Results were collected, tabulated, and statistically analyzed using statistical package SPSS, version 17.0 (SPSS Inc., Chicago, Illinois, USA).


  Results Top


In HCC patients, on comparing different tumor characters it was found that 75.5% of HCC patients had tumor size more than 32 m, and 34.7% of them had multiple focal lesions.

According to the site of the tumor, right hepatic lobe lesion was found in 65.3%, whereas 14.3% of patients had both right and left lobe lesions. Blood vessels were patent in 53.1% and thrombosed in 46.9% of HCC patients. According to the Barcelona Clinic Liver Cancer staging system, 6.1% of patients were of category A, 22.4% were of category B, 18.4% were of category C, and 53.1% were of category D [Table 1].
Table 1: Tumor characters in the hepatocellular carcinoma group

Click here to view


On comparing carriers of the three genotypes (5G/5G, 4G/5G, and 4G/4G) as regards sex, Child score, age, total bilirubin, direct bilirubin, albumin, serum glutamate pyruvate transaminase, alkaline phosphatase, γ-glutamyltranspeptidase, international normalized ratio, prothrombin, and α-fetoprotein, there were no statistically significant differences among them (P = 0.81, 0.76, 0.46, 0.054, 0.99, 0.92, 0.22, 0.58, 0.49, 0.42, 0.54, and 0.85, respectively). On comparing the three genotypes (5G/5G, 4G/5G, and 4G/4G) among HCC patients as regards serum glutamic oxaloacetic transaminase, there was a statistically significant difference (P = 0.02) [Table 2].
Table 2: Genotypes in relation to different studied parameters among hepatocellular carcinoma patients

Click here to view


On comparing carriers of the three genotypes (5G/5G, 4G/5G, and 4G/4G) as regards different tumor characters (tumor size, tumor number, tumor site, and Barcelona Clinic Liver Cancer staging system), there were no significant differences among them (P = 0.16, 0.59, 0.42, and 0.14, respectively).

Rate of thrombosis in the HCC group was 62.5% in 4G/5G genotype versus 41.2 and 12.5% in 5G/5G and 4G/5G genotypes, respectively; there was a statistically significant difference (P = 0.04) [Table 3].
Table 3: Genotypes in relation to different tumor characters among hepatocellular carcinoma patients

Click here to view



  Discussion Top


This work aimed to investigate the distribution of genotypes and the frequency of alleles of SERPINE1/PAI-1 4G/5G polymorphism in patients with HCC.

In the present study, we found that the distribution of different genotypes of PAI-1 4G/5G polymorphism among HCC patients was as follows: 49% had 4G/5G genotype, 34.7% had 5G/5G, and 16.3% had 4G/4G.

Divella et al. [9] found that the frequency of the 4G allele and the 4G/4G SERPINE1 genotype was significantly higher in patients with HCC associated with viral infection than in patients without viral infection.

Espino et al. [10] compared the genotype of patients with liver fibrosis and those without fibrosis; no significant differences were observed in the genotype of the 4G/5G polymorphism of PAI-1 gene or in the allele distribution. Moreover, they did not find differences in the genotype or allele distribution when comparing between patients with histological criteria of nonalcoholic steatohepatitis associated with fibrosis and patients without nonalcoholic steatohepatitis. We studied the influence of SERPINE1 4G/5G polymorphism on the clinicopathological profile in patients with HCC. We investigated the association of the PAI-1 4G/5G polymorphism with demographic parameters (age and sex), laboratory parameters (liver function), and Child score in HCC patients. We did not identify any association between the PAI-1 4G/5G polymorphism and all studied parameters.

In this study, HCC was more prevalent in male patients than in female patients with a ratio of 3.9/1 (male patients accounted for 79.6% and female patients accounted for 20.4%). This is in agreement with the findings of Huo et al. [11], Hsu et al. [12], and Lee et al. [13], who found that the prevalence of male HCC patients was 73, 77, and 78%, respectively. HCC has a strong male predominance, with a male-to-female ratio estimated to be 2.4/1 [14].

We found that genotype 4G/5G was significantly increased in thrombosed blood vessels. These results are in agreement with those of Aĭsina et al. [15], who showed that arterial and to a greater extent venous thromboses were associated with the 4G/5G polymorphism of PAI-1 gene and high plasma level of the inhibitor in 79% of antiphospholipid syndrome patients. Moreover, Wang et al. [16] demonstrated the role of PAI-1 4G/5G polymorphism being a risk candidate locus for venous thromboembolism susceptibility, especially in patients with other genetic thrombophilic disorders. Similarly, Wiklund et al. [17] suggested a true and clinically important association between PAI-1 4G/5G genotype and risk for future ischemic stroke.


  Conclusion Top


PAI-1 4G/5G polymorphism was not associated with HCC.

Apart from a significant increase in 4G/5G genotype among HCC patients with thrombosed portal vein than in those with patent vein, there was no significant association between PAI-1 4G/5G polymorphism and the other studied clinicopathological parameters in HCC patients.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

 
  References Top

1.
Altekruse SF, McGlynn KA, Reichman ME. Hepatocellular carcinoma incidence, mortality, and survival trends in the United States from 1975 to 2005. J Clin Oncol 2009; 27:1485–1491.  Back to cited text no. 1
    
2.
El-Bassuonia MA, El-Edel RH, Gawesh EM, Mandour SS. The diagnostic value of serum squamous cell carcinoma antigen for prediction of hepatocellular carcinoma. Menoufia Med J 2016; 29:141–146.  Back to cited text no. 2
    
3.
Elsabawy MN. Hepatitis gender gap in Egypt: a study in medical geography. Procedia Soc Behav Sci 2011; 19:121–130.  Back to cited text no. 3
    
4.
Jones RG, Thompson CB. Tumor suppressors and cell metabolism: a recipe for cancer growth. Genes Dev 2009; 23:537–548.  Back to cited text no. 4
    
5.
Dupont DM, Madsen JB, Kristensen T, Bodker JS, Blouse GE, Wind T, et al. Biochemical properties of plasminogen activator inhibitor-1. Front Biosci 2009; 14:1337–1361.  Back to cited text no. 5
    
6.
Binder BR, Mihaly J, Prager GW. uPAR-uPA-PAI-1 interactions and signaling: a vascular biologist's view. Thromb Haemost 2007; 97:336–342.  Back to cited text no. 6
    
7.
Chorostowska-Wynimko J, Skrzypczak-Jankun E, Jankun J. Plasminogen activator inhibitor type-1: its structure, biological activity and role in tumorigenesis (Review). Int J Mol Med 2004; 13:759–766.  Back to cited text no. 7
    
8.
Shih CM, Kuo WH, Lin CW, Chen W, Cheng WE, Chen SC, Lee YL Association of polymorphisms in the genes of the urokinase plasminogen activation system with susceptibility to and severity of non-small cell lung cancer. Clin Chim Acta 2011; 412:194–198.  Back to cited text no. 8
    
9.
Divella R, Mazzocca A, Gadaleta C, Simone G, Paradiso A, Quaranta M, Daniele A. Influence of plasminogen activator inhibitor-1 (SERPINE1) 4G/5G polymorphism on circulating SERPINE-1 antigen expression in HCC associated with viral infection. Cancer Genomics Proteomics 2012; 9:193–198.  Back to cited text no. 9
    
10.
Espino A, Villagrán A, Vollrath V, Hanckes P, Salas R, Farah A, et al. Plasminogen activator inhibitor type 1 serum levels and 4G/5G gene polymorphism in morbidly obese Hispanic patients with non-alcoholic fatty liver disease. Ann Hepatol 2011; 10:493–501.  Back to cited text no. 10
    
11.
Huo TI, Hsu CY, Huang YH, Su CW, Lin HC, Lee RC, et al. Prognostic prediction across a gradient of total tumor volume in patients with hepatocellular carcinoma undergoing locoregional therapy. BMC Gastroenterol 2010; 10:146.  Back to cited text no. 11
    
12.
Hsu CY, Hsia CY, Huang YH, Su CW, Lin HC, Lee PC, et al. Selecting an optimal staging system for hepatocellular carcinoma: comparison of 5 currently used prognostic models. Cancer 2010; 116:3006–3014.  Back to cited text no. 12
    
13.
Lee SS, Shin HS, Kim HJ, Lee SJ, Lee HS, Hyun KH, et al. Analysis of prognostic factors and 5-year survival rate in patients with hepatocellular carcinoma: a single-center experience. Korean J Hepatol 2012; 18:48–55.  Back to cited text no. 13
    
14.
International Agency for Research on Cancer (IARC). Combined estrogen-progestogen contraceptives and combined estrogen-progestogen menopausal therapy. IARC Monogr Eval Carcinog Risks Hum 2007; 91:1–528.  Back to cited text no. 14
    
15.
Aĭsina RB, Mukhametova LI, Ostriakova EV, Seredavkina NV, Patrushev LI, Patrusheva NL, et al. Polymorphism of the plasminogen activator inhibitor type 1 gene, plasminogen level and thrombosis in patients with antiphospholipid syndrome. Biomed Khim. 2014; 60:72–93.  Back to cited text no. 15
    
16.
Wang J, Wang C, Chen N, Shu C, Guo X, He Y, Zhou Y. Association between the plasminogen activator inhibitor-1 4G/5G polymorphism and risk of venous thromboembolism: a meta-analysis. Thromb Res 2014; 134:1241–1248.  Back to cited text no. 16
    
17.
Wiklund PG, Nilsson L, Ardnor SN, Eriksson P, Johansson L, Stegmayr B, et al. Plasminogen activator inhibitor-1 4G/5G polymorphism and risk of stroke: replicated findings in two nested case-control studies based on independent cohorts. Stroke 2005; 36:1661–1665.  Back to cited text no. 17
    


    Figures

  [Figure 1], [Figure 2]
 
 
    Tables

  [Table 1], [Table 2], [Table 3]



 

Top
 
 
  Search
 
Similar in PUBMED
   Search Pubmed for
   Search in Google Scholar for
 Related articles
Access Statistics
Email Alert *
Add to My List *
* Registration required (free)

 
  In this article
Abstract
Introduction
Patients and Methods
Results
Discussion
Conclusion
References
Article Figures
Article Tables

 Article Access Statistics
    Viewed686    
    Printed8    
    Emailed0    
    PDF Downloaded56    
    Comments [Add]    

Recommend this journal


[TAG2]
[TAG3]
[TAG4]