Manipulation of Interleukin-6 (IL-6) and Transforming Growth Factor Beta-1(TGFβ-1) towards viral induced liver cancer pathogenesis

Abstract

Background: Hepatocellular carcinoma (HCC) is the most common liver malignancy. Early diagnosis of HCC has always been challenging. Hepatitis-C Virus (HCV)- induced HCC makes up for 25% of HCC cases annually. Many studies have reported the association of TGFβ-1 and IL-6 gene polymorphisms with the predisposition of HCC. This study aims to assess the pathogenicity and the prevalence of IL-6 -174G/C (rs1800795) and TGFβ-1 +29C/T (rs1800470) polymorphisms in HCV-infected HCC patients. Methodology: This study uses multiple bioinformatics tools to analyse the pathogenicity of the TGFβ-1 +29 C/T and IL-6 -174 G/C polymorphisms. AliBaba2 was used to predict the transcription factor binding sites in the mutant and wild type IL 6 genes. The structural changes in the mutant TGFβ-1 structure were determined through project HOPE. To assess the polymorphic prevalence of IL-6 -174 G/C and TGFβ-1 +29 C/T genotypes in HCC and control subjects, ARMS-PCR was performed on 213 diseased and 216 control samples. GraphPad Prism was used for the statistical analysis of the results. CB Dock was used for the molecular docking analysis of tetrahydroxyflavanone with IL-6 and TGFβ-1 proteins to predict its inhibitory potential against the two cytokines. Results: In-silico analysis revealed the regulatory nature of both IL-6 -174 G/C and TGFβ-1 +29 C/T polymorphisms but with limited functional significance. The structural variation in mutant TGFβ-1 such as Pro10Leu substitution might contribute to the structural instability. An additional transcription factor binding site of nuclear factor-1 was identified in the mutant IL-6 structure. ARMS-PCR results revealed that the individuals carrying TT genotype for TGFβ-1 gene have an increased risk of developing HCC (p<0.0001, OR=5.403, RR=2.062) as compared to individuals with CT and CC genotype. Similarly, GC genotype carriers for IL-6 gene exhibit an increased risk of HCC susceptibility (p<0.0001, OR=2.276, RR=1.512) as compared to the people carrying the GG genotype. The molecular docking results revealed considerable docking efficiency of tetrahydroxyflavanone with IL-6 (-7.3 kJ/mol) and TGFβ-1 (-7.7 kJ/mol) proteins. Compared to the conventional chemotherapy, tetrahydroxyflavanonone shows a promising potential for treating HCC. Conclusion: The TT genotype of TGFβ-1 gene and the GC genotype of IL-6 gene are found to be associated with the development of HCC in patients. Tetrahyrdoxyflavanone is proposed as a promising treatment option for HCC.