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Tumor necrosis factor- α -G308A polymorphism is associated with liver pathological changes in hepatitis C virus patients

View Article: PubMed Central - PubMed

ABSTRACT

Aim: To investigate the association of tumor necrosis factor alpha (TNFα) -G308A polymorphism with different liver pathological changes in treatment-naïve Egyptian patients infected with hepatitis C virus (HCV) genotype 4.

Methods: This study included 180 subjects, composed of 120 treatment-naïve chronic HCV patients with different fibrosis grades (F0-F4) and 60 healthy controls. The TNFα -G308A region was amplified by PCR and the different genotypes were detected by restriction fragment length polymorphism analysis. The TNFα protein was detected by enzyme-linked immunosorbent assay. The influence of different TNFα -G308A genotypes on TNFα expression and liver disease progression were statistically analyzed. The OR and 95%CI were calculated to assess the relative risk confidence.

Results: Current data showed that the TNFα -G308A SNP frequency was significantly different between controls and HCV infected patients (P = 0.001). Both the AA genotype and A allele were significantly higher in late fibrosis patients (F2-F4, n = 60) than in early fibrosis patients (F0-F1, n = 60) (P = 0.05, 0.04 respectively). Moreover, the GA or AA genotypes increased the TNFα serum level greater than the GG genotype (P = 0.002). The results showed a clear association between severe liver pathological conditions (inflammation, steatosis and fibrosis) and (GA + AA) genotypes (P = 0.035, 0.03, 0.04 respectively). The stepwise logistic regression analysis showed that the TNFα genotypes (GA + AA) were significantly associated with liver inflammation (OR = 3.776, 95%CI: 1.399-10.194, P = 0.009), severe steatosis (OR = 4.49, 95%CI: 1.441-14.0, P = 0.010) and fibrosis progression (OR = 2.84, 95%CI: 1.080-7.472, P = 0.034). Also, the A allele was an independent risk factor for liver inflammation (P = 0.003), steatosis (P = 0.003) and fibrosis (P = 0.014).

Conclusion: TNFα SNP at nucleotide -308 represents an important genetic marker that can be used for the prognosis of different liver pathological changes in HCV infected patients

No MeSH data available.


Related in: MedlinePlus

Tumor necrosis factor α -G308A polymorphism analysis. A: TNFα -308 PCR-RFLP analysis, genomic DNA was extracted, amplified by PCR, digested with NcoI restriction enzyme and run on a 4% agarose gel. Lanes 1, 2, 3 and 4 correspond to PCR products before digestion, GG homozygote (87 bp), AA homozygote (107 bp) and GA heterozygote (107 and 87 bp) respectively. B: TNFα -308 PCR sequence analysis, the PCR products of different TNFα -308 genotypes were purified and sequenced using the reverse primer. The NcoI restriction site is highlighted and the arrow points to the single nucleotide polymorphism. TNFα: Tumor necrosis factor alpha; HCV: Hepatitis C virus; RFLP: Restriction fragment length polymorphism.
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Figure 1: Tumor necrosis factor α -G308A polymorphism analysis. A: TNFα -308 PCR-RFLP analysis, genomic DNA was extracted, amplified by PCR, digested with NcoI restriction enzyme and run on a 4% agarose gel. Lanes 1, 2, 3 and 4 correspond to PCR products before digestion, GG homozygote (87 bp), AA homozygote (107 bp) and GA heterozygote (107 and 87 bp) respectively. B: TNFα -308 PCR sequence analysis, the PCR products of different TNFα -308 genotypes were purified and sequenced using the reverse primer. The NcoI restriction site is highlighted and the arrow points to the single nucleotide polymorphism. TNFα: Tumor necrosis factor alpha; HCV: Hepatitis C virus; RFLP: Restriction fragment length polymorphism.

Mentions: The amplified TNFα -308 PCR products were digested with NcoI restriction enzyme, and run on 4% agarose gel, as shown in Figure 1A. The homozygote AA genotype at TNFα -308 showed the original 107 bp fragment intact (lacking the NcoI site), while the homozygote GG genotype showed two fragments of 87 and 20 bp. The heterozygote GA genotype showed three fragments of 87, 20 and 107 bp. Moreover, the sequence results confirmed the integrity of the NcoI restriction site’s surrounding area and verified the results of the TNFα -308 PCR-RFLP analysis. The sequence results for the TNFα -308 different genotypes are shown in Figure 1B.


Tumor necrosis factor- α -G308A polymorphism is associated with liver pathological changes in hepatitis C virus patients
Tumor necrosis factor α -G308A polymorphism analysis. A: TNFα -308 PCR-RFLP analysis, genomic DNA was extracted, amplified by PCR, digested with NcoI restriction enzyme and run on a 4% agarose gel. Lanes 1, 2, 3 and 4 correspond to PCR products before digestion, GG homozygote (87 bp), AA homozygote (107 bp) and GA heterozygote (107 and 87 bp) respectively. B: TNFα -308 PCR sequence analysis, the PCR products of different TNFα -308 genotypes were purified and sequenced using the reverse primer. The NcoI restriction site is highlighted and the arrow points to the single nucleotide polymorphism. TNFα: Tumor necrosis factor alpha; HCV: Hepatitis C virus; RFLP: Restriction fragment length polymorphism.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5016377&req=5

Figure 1: Tumor necrosis factor α -G308A polymorphism analysis. A: TNFα -308 PCR-RFLP analysis, genomic DNA was extracted, amplified by PCR, digested with NcoI restriction enzyme and run on a 4% agarose gel. Lanes 1, 2, 3 and 4 correspond to PCR products before digestion, GG homozygote (87 bp), AA homozygote (107 bp) and GA heterozygote (107 and 87 bp) respectively. B: TNFα -308 PCR sequence analysis, the PCR products of different TNFα -308 genotypes were purified and sequenced using the reverse primer. The NcoI restriction site is highlighted and the arrow points to the single nucleotide polymorphism. TNFα: Tumor necrosis factor alpha; HCV: Hepatitis C virus; RFLP: Restriction fragment length polymorphism.
Mentions: The amplified TNFα -308 PCR products were digested with NcoI restriction enzyme, and run on 4% agarose gel, as shown in Figure 1A. The homozygote AA genotype at TNFα -308 showed the original 107 bp fragment intact (lacking the NcoI site), while the homozygote GG genotype showed two fragments of 87 and 20 bp. The heterozygote GA genotype showed three fragments of 87, 20 and 107 bp. Moreover, the sequence results confirmed the integrity of the NcoI restriction site’s surrounding area and verified the results of the TNFα -308 PCR-RFLP analysis. The sequence results for the TNFα -308 different genotypes are shown in Figure 1B.

View Article: PubMed Central - PubMed

ABSTRACT

Aim: To investigate the association of tumor necrosis factor alpha (TNFα) -G308A polymorphism with different liver pathological changes in treatment-naïve Egyptian patients infected with hepatitis C virus (HCV) genotype 4.

Methods: This study included 180 subjects, composed of 120 treatment-naïve chronic HCV patients with different fibrosis grades (F0-F4) and 60 healthy controls. The TNFα -G308A region was amplified by PCR and the different genotypes were detected by restriction fragment length polymorphism analysis. The TNFα protein was detected by enzyme-linked immunosorbent assay. The influence of different TNFα -G308A genotypes on TNFα expression and liver disease progression were statistically analyzed. The OR and 95%CI were calculated to assess the relative risk confidence.

Results: Current data showed that the TNFα -G308A SNP frequency was significantly different between controls and HCV infected patients (P = 0.001). Both the AA genotype and A allele were significantly higher in late fibrosis patients (F2-F4, n = 60) than in early fibrosis patients (F0-F1, n = 60) (P = 0.05, 0.04 respectively). Moreover, the GA or AA genotypes increased the TNFα serum level greater than the GG genotype (P = 0.002). The results showed a clear association between severe liver pathological conditions (inflammation, steatosis and fibrosis) and (GA + AA) genotypes (P = 0.035, 0.03, 0.04 respectively). The stepwise logistic regression analysis showed that the TNFα genotypes (GA + AA) were significantly associated with liver inflammation (OR = 3.776, 95%CI: 1.399-10.194, P = 0.009), severe steatosis (OR = 4.49, 95%CI: 1.441-14.0, P = 0.010) and fibrosis progression (OR = 2.84, 95%CI: 1.080-7.472, P = 0.034). Also, the A allele was an independent risk factor for liver inflammation (P = 0.003), steatosis (P = 0.003) and fibrosis (P = 0.014).

Conclusion: TNFα SNP at nucleotide -308 represents an important genetic marker that can be used for the prognosis of different liver pathological changes in HCV infected patients

No MeSH data available.


Related in: MedlinePlus