Liquid Biopsy-Based Profiling of TP53 Gene Hotspot Mutations in HBV-Induced Chronic Liver Disease and Hepatocellular Carcinoma Patients: A Study From Bangladesh.

Abstract

Background: The tumor protein (TP53) gene is a common driver gene for somatic mutations in hepatocellular carcinoma(HCC), and detection may provide insight into evaluating disease progression. Circulating cell-free DNA (cfDNA) from liquid biopsy has emerged as a promising noninvasive biopsy material for identifying potential genetic alterations. This study explored the mutational signature of the TP53 gene in Hepatitis B virus (HBV)-infected chronic liver disease (CLD) and HCC patients from cfDNA.

Method: A total of 80 participants: 30 HCC, 30 CLD, and 20 healthy controls were included in this cross-sectional study. cfDNA was extracted from aseptically collected peripheral venous blood. TP53 (R249S) gene was amplified using selected primers via conventional PCR and visualized on a 2% agarose gel electrophoresis. Amplified products underwent Sanger sequencing; mutational analysis was done using MEGA11 software compared with the reference sequence.

Result: TP53 hotspot mutations were found in 23.3% (7/30) of HCC patients. Mutational frequencies were detected: 13.3% (4/30) at position 746G > A (R249S), 6.7% (2/30) at 747G > A (R249S), and 3.3% (1/30) at 735C > T (G245S). Among CLD patients, a single hotspot mutation (3.3%) was detected at position 744G > A/T(R248Q/W). However, no hotspot mutation was detected in healthy controls. All HCC patients with mutations (7/7, 100%) had elevated ALT, AST, serum bilirubin, and AFP levels, and in the unmutated (wild) group, these were 69.6%, 47.8%, 30.4%, and 60.9%, respectively. The mean AST and AFP levels were significantly higher in mutated patients compared with the wild group.

Conclusion: Detection of TP53 gene mutations along with biochemical markers in the management of HCC is evident, and it may play an important role in targeted therapy and personalized treatment plans.