| [Objective]Primary hepatolcellular carcinoma(HCC) is one of the highest morbidity of malignant tumors at present. Epidemiology has confirmed that the infection of HBV is the independent risk factor of HCC. Covalently closed circular DNA is an important replicative intermediate in the life period of HBV, which has been generally considered as the basic reason of persistent viral infection and liver progressive pathological changes. Many studies in vitro and animal reseachers have shown that there is a closely relationship between the HBVcccDNA and hepatocellular carcinoma. But there are less reports about the relationship between HBVcccDNA from cancerous tissues of patients and the development of HCC.To explore the quantity of HBVcccDNA and total HBVDNA in cancerous tissues and non-cancerous tissues from patients with HCC by FQ-PCR. To illuminate the relationship between the HBVcccDNA and the criterion of HCC and liver damage through evaluating the effect of HBVcccDNA in the development of HCC.[Methods]Detected 45 patients with HCC(including 36 men and 9 women) who were in Shandong Provincial Hospital since December 2006 to May 2007. There were 42 with HBsAg-positive, 3 with HBsAg-negtive including 2 with HBeAb-positive and 1 with HBcAb-positive. Anti-HCV was all negative. All patients were not given antiviral treatment. The diagnose of HCC were identified with pathology. Cancerous tissues and non-cancerous tissues are obtained from liver operation. There was no cancerous cells on the cutting edge. Part of sample was used to evalue the criterion of HCC, another was used to detect the quantity of HBVcccDNA and total HBVDNA by FQ-PCR. Hepatic function index such as ALT,TBIL,PT was detected with the automatic biochemistry analyzer.[Results]1,HBVcccDNA was detected in all patients. For 42 patients of HBsAg-positive, HBVcccDNA of cancerous tissues was quantified between 3.22×10~1 copies/mg to 4.51×10~4 copies/mg (logarithm value: 3.09±1.22), and the total HBVDNA between 1.67×10~2 copies/mg to 6.43×10~5 copies/mg (logarithm value: 4.33±1.11) ; HBVcccDNA of non-cancerous tissues was quantified between 1.17×10~1 copies/mg to 5.95×10~4 copies/mg (logarithm value: 2.62±0.97) , and the total HBVDNA between 1.42×10~2 copies/mg to 1.26×10~6 copies/mg (logarithm value: 4.13±1.09).2,For sera HBsAg-positive patients , the total HBVDNA levels in cancerous tissues and non-cancerous tissues were comparable (P = 0.439) .But HBVcccDNA was significantly up regulated in the cancerous tissues (3.09±1.22 vs 2.62±0.97, P= 0.039)3,For sera HBsAg-positive patients, the cancerous tissues had a significantly higher proportion of total HBVDNA in the form of cccDNA (P=0.015) .Fourteen out of 42(14/42,33%) cancerous tissues of HBsAg-positive patiens had HBVDNA solely in cccDNA form nearly. 4,There was a significant positive correlation between the quantity of HBVcccDNA and total HBVDNA in cancerous tissues (r=0.8244,P<0.001). A same result was also found in non-cancerous tissues (r=0.8460, P<0.001) .5,HBVcccDNA was also detected in the tissues of 3 HBsAg-negative patiens .The proportion of cccDNA in tumor was higher than in non-cancerous tissues.6,There was no well-defined correlation between quantity of HBVcccDNA in the cancerous tissues and the criterion of hepatocellular carcinoma (P>0.05) . 7,There was no well-defined correlation between quantity of HBVcccDNA and ALT,TBIL,PT (P>0.05) .[Conclusion]1,The proportion of HB VcccDNA in cancerous tissues was higher than innon-cancerous tissues.2,HB VcccDNA was detected in the tissues of HBsAg-negative patiens with HCC,Cancerous tissues of these patients had HBVDNA solely in cccDNA form nearly.3,The cancerous tissues had significantly higher levels of HB VcccDNA, while theintrahepatic total HBVDNA in cancerous tissues and non-cancerous tissues werecomparable.4,There was no well-defined correlation between quantity of HB VcccDNA anddegree of liver parenchyma injury, which suggested the level of HBVcccDNA incancerous tissues could't act as a predictive index of the malignant degree of HCC. |
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