p53 tumor suppressor gene mutations in hepatocellular carcinoma patients in India

BACKGROUND, Specific mutations of the p53 tumor suppressor gene in hepatoce llular carcinoma (HCC) have been reported from several parts of the world, bur to the authors' knowledge to date the status of this gene has not been studied in HCC patients in India, where HCC is one of the major cancers and the frequency of chronic hepatitis B virus (HBV) as well as hepatitis C vi rus (HCV) infection and exposure to dietary aflatoxin B, is very high. The most frequent mutation of the p53 gene in HCC is an AGG(Arg) to AGT(Ser) mi ssense mutation at codon 249 of exon 7. METHODS. Liver biopsy specimens from 21 HCC patients and 10 healthy control s were obtained through surgery or by needle biopsy technique. Phenol-chlor oform-extracted DNA specimens were employed for the detection of HBV infect ion and p53 gene mutations. Nucleotide mutations of exons 4-9 of the p53 ge ne were analyzed by polymerase chain reaction (PCR), single strand confirma tion polymorphism, and direct sequencing. Third-generation sandwich enzyme- linked immunosorbent assay (ELISA) was used for the serologic detection of HBV and HCV infection. RESULTS. Analysis of exons 4-9 of the p53 gene revealed only 3 mutations (3 of 21 specimens, 14.28%; 95% confidence interval, -0.7-29.3), 2 mutations at codon 249 showing G-->T transversions, and 1 mutation (4.7%) at codon 25 0 with a C-->T transition. The base substitutions at the third base of codo n 249 resulted in a missense mutation leading to a change in amino acid fro m arginine to serine whereas at codon 250 it caused a change from proline t o serine. Dot blot hybridization and PCR for HBV DNA from HCCs revealed 58. 8% (10 of 17 specimens) and 90.47% (19 of 21 specimens), positivity, respec tively. ELISA for hepatitis B virus surface antigen in serum showed a posit ivity of 71.42% (15 of 21 specimens), but there was only 40% positivity (8 of 20 specimens) for hepatitis B virus envelope antigen whereas 6 of 17 pat ients (35.29%) showed the presence of antibodies against hepatitis B virus envelope protein. No patient was found to be positive for the HCV antibody. CONCLUSIONS. The very low frequency of p53 mutations and the extremely high frequency of HBV infection (> 90%) in HCC indicate that the mutations in t he p53 gene frequently found in HCC reported from different endemic areas o f the world may not play a direct role in the development of HCC in India. HBV infection and, possibly, exposure to the dietary aflatoxin B-1 appear t o play major roles in the molecular pathogenesis of HCC in India. (C) 2000 American Cancer Society.