The Effects Of HBeAg And Mitochondrial DNA D-Loop Region Variations In Hepatocellular Carcinoma

Objective To explore the relationship between the serum HBeAg and Mitochondrial DNA (mtDNA) D-Loop region variations and their effects in hepatocellular carcinoma.Methods 40 cases with hepatocellular carcinoma (HCC) who were consequentially hospitalized for radical resection of HCC in the department of hepatobiliary surgery of the first affiliated hospital to Guangxi medical university from January 2007 to December 2007 were selected randomly,and their cancer tissue and non-cancer tissue were all studied.Additionally,8 patients who underwent liver trauma without any liver desease were selected as normal liver tissue.Polymerase chain reaction (PCR) and direct sequencing were applied to study the mtDNA D-Loop region,and enzyme linked immunosorbent assay (ELISA) was applied to detect the serum HBeAg.Results The sequences of cancer tissue ,non-cancer tissue and normal liver tissue were compared with the sequences of the mtDNA databank using DNAstar software.1. mtDNA D-Loop region was very variable with 123 nucleotide sites variations (11.0%,123/1 122) in our study. 2. Two variations were considered as new polymorphism who were not recorded in data bank.They were nucleotide (NT) 228 G→C and NT 16 092 T→A.3. There were 34 mutational sites in our study with 16 sites of point mutation,5 sites of insertion,and 13 sites of deletion.There were two main kinds of point mutation in our study,and they were T→C and G→A with a percentage of 43.8%(7/16).4. The mutation rates of mtDNA D-Loop region were 40.0%(16/40) in cancer tissue, 10.0%(4/40) in non-cancer tissue,and 0.0%(0/8) in normal tissue.There was a significant difference among them (P<0.05).5. Of 40 cases with HCC,30 cases were HBeAg positive (60.3%,25/40),and 10 cases were also with mtDNA D-Loop region mutation.There was a relationship between mtDNA mutation and HBeAg positive with a high mtDNA D-Loop mutation rate in cancer tissue with HBeAg positive (P<0.05).Conclusions1. The mtDNA D-Loop region is a region with a high rate of variation and mutation, especially in HCC tissue.2. The mutation rate of mtDNA D-Loop region in non-cancer tissue of HCC is 10.0%, which implies that it is the molecule appearance of theprecancerosis of HCC.3. There is a high mtDNA D-Loop mutation rate in HCC tissues with HBeAg positive .