| Objective:through studying the mutation and its protein’s expression level of gene p53 under the bi-exposure condition of hepatitis B virus (HBV) and aflatoxin B1 (AFB1), to explore the relationship between gene p53, under bi-exposure condition, and the occurrence and development of hepatocelluar carcinoma (HCC).Methods:82 cases, from different areas of Guangxi province, were categorized into HB V(+)/AFB 1 (+) (experimental group A),44 cases; HBV(+)/AFB1(-) (control group B),15 cases; HBV(-)/AFB1(+) (control group C),15 cases; and HBV(-)/AFB1(-) (control group D),8 cases, according to the exposure condition of HBV and AFB1. And for normal controls,20 cases of hepatic hemangioma liver resection and liver transplant donor hepatic tissue were selected, including HBV(-)/AFB 1 (+) (normal hepatic tissue control group E),9 cases,and HBV (-)/AFB1 (-) (normal hepatic tissue control group F),11 cases. By means of PCR direct sequenced and immunohistochemistry (IHC), the mutation of 11 pairs’ exons of gene p53 and the expression condition of the protein were tested respectively.Results:(1) The total mutation rate of gene P53 in 82 HCC cases was 40.2%, (33/82); the hot point of gene P53 mutation was 249 coding region, occupied 86.8%(33/38)of the total mutation rate; the site mutations out of the 249 coding region took up 13.2%(5/38)of the total mutation rate. In P53exon7 249, G→T mutation rate was 90.9%(30/33), G→C mutation rate was 9.1%(3/33). The positive mutation rate of gene P53exon7 249 was 40.2%(33/82) in 82 cases, among them, the positive rate of experimental group A, control group B, C and D was 50%(22/44),13.3% (2/15),46.7%(7/15) and 25%(2/8) respectively. Overall comparison among experiment group A and control groups B, C and D, the difference was with no statistical significance (P>0.05); through comparison between groups, between experiment group A and control group B, experiment group A was with higher mutation rate (P<0.05), the differences among rest groups were not significant (P>0.05). (2) In the expression of gene p53, the positive rate in 82 cases was 74.4%(61/82), and the positive rate in experiment group A, control group B, C and D was 90.9%(40/44), 46.7%(7/15),73.3%(11/15) and 37.5%(3/8) respectively. The IOD values were:experiment group A (1083.8412±1554.3369), control group B (376.9396±343.64576), control group C (795.9503±93075329), control group D (148.1998±42.8268), and normal hepatic tissue control group (63.4710±33.83466). Compared with the positive expression rate of gene p53 protein, overall comparison among experiment group A and control groups B, C and D, experiment group A was with higher positive expression rate (P<0.05); compared between groups, experiment group A compared with control group B and D, both higher expression rate could be seen in experiment group A (P<0.05), the differences among the rest groups were with no statistical significance (P>0.05). In the IOD value of gene p53 IHC, overall comparison among experiment group A and control group B, C and D, experiment group A had a higher IOD value (P<0.05), through comparisons between each two groups, between expefiment group A and control group D, experiment group A took a higher IOD value (P<0.05), between control group B and D, control group B was with a higher IOD value (P<0.05), between control group C and D, higher IOD value (P<0.05)could be seen in control group C, the differences were with no statistical significance (P>0.05) in the comparison between rest groups. (3) The positive cases of P53exon7 249 mutation and p53 protein expression were both 29. There were 32 cases, which were positive in P53exon7 249 mutation but negative in p53 protein expression. And 4 cases were negative in P53exon7 249 mutation but positive in p53 protein expression. P53exon7 349 mutation and P53 protein expression presented correlation generally (P<0.05); analyzed the comparison between groups, there was certain correlation (P<0.05) between P53exon7 249 mutation and P53 protein expression; between control groups, the correlation of P53exon7 249 mutation and P53 protein expression were all with no statistical significance (P>0.05).Conclusions:Under the bi-exposure condition of HBV/AFB1 in Guangxi province, the mutation and expression of P53 gene in hepatocellular carcinoma were with high positive rate, the P53exon7 249 mutation was its mutation hot point, which is a common molecular biological event in this district; the high mutation rate and expression rate of gene P53 could relate to the synergy of HB V/AFB 1 bi-exposure factor in this district. |
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