Hepatitis B virus (HBV) genotype C is a leading risk factor for hepatocellular carcinoma (HCC). Eight HBV isolates from Sohag governorate, Egypt were confirmed by polymerase chain reaction (PCR), resulted in 1110 nt correspond to partial sequence of polymerase (pol) and envelop (env) genesTwo HBV isolates were associated with HCC (Sohag1 and 7). Phylogenetic analysis of the nucleotide sequence and the amino acid of the pol or env genes revealed that 7 out of the 8 isolates were closely related to genotype D. Only Sohag 1-HCC isolate was clustered with genotype C which is known as risk factor of HCC. Multiple sequence alignment resulted in unique amino acid substitutions (V320→L and N464→T) and (I284→L, T300→I and K334→R) in pol and env genes respectively. Further prospective studies are needed to confirm the role of these mutations in the development of HCC.