【摘要】 目的分析慢性乙型肝炎肝硬化结节样变与HBV变异的关系以及HBV变异在慢性肝硬化、肝癌进展中的作用。方法收集临床诊断慢性乙型肝炎肝硬化患者104例,其中不典型结节样增生病例43例,单纯肝硬化患者41例,肝癌患者20例。采用荧光探针实时定量PCR试剂盒提取HBV基因组,选取PCR强阳性产物用Sanger双脱氧链末端终止法对HBV前C区变异基因G1896A,BCP区G1764A、A1762T位点在全自动核苷酸分析仪上测序,测序结果与Gene Bank中pADR标准株序列作对比分析。结果结节性增生组、单纯肝硬化组、肝癌组患者HBV前C Gl896A变异检出率分别是52.3%、40.1%和37.4%,差异无统计学意义(χ2=0.547,P=0.05);结节性增生组BCP A l762T变异率68.4%,与单纯硬化组36.3%比较,差异有统计学意义(P=0.038),与肝癌组G1764A变异检出率分别是73.0%,与对照组比较具有显著性差异(P=0.0411)。单纯肝硬化组、结节性增生组BCP基因变异(+)组,HBV DNA载量2.18×107、1.2×106高于基因变异(-)组1.18×104、2.95×103,差别有统计学意义(P=0.0451,P=0.0412);结节性增生组、肝癌组BCP基因变异(+)组HBeAg定量750.00IU/mL,1300.00IU/mL高于基因变异(-)组416.13 IU/mL,927.60 IU/mL差别有统计学意义(P=0.0451,P=0.0073)。结论前C区变异与乙肝肝硬化结节样变临床进展为肝癌无关,而BCP区变异与乙肝结节样变临床进展为肝癌有关。更多还原

【Abstract】 Objective To analyze the relationship between hepatic dysplastic nodules(HDN)from HBV-related cirrhosis and HBV mutation.Methods One hundred and four cases of clinically diagnosed as HBV-related cirrhosis were enrolled,43(HDN group)of which had a history of liver cirrhosis(LC)with abdominal ultrasound or CT diagnosis of atypical nodular hyperplasia,41(LC group)of which were cirrhosis patients without nodular hyperplasia,and 20(hepatocellular carcinoma,HCC group)of which were liver cancer patients.HBV genome was exacted by real-time quantitative polymerase chain reaction(PCR)using a fluorescent probe extraction reagent kit.Strong positive PCR product was sequenced with Sanger dideoxy chain termination method to examine HBV mutations on pre-C G1896A and BCP G1764A,A1762T.Sequencing results were compared with pADR standard strain sequence of gene bank.Results The HBV mutation detection rate on pre-C G1896 A was 52.3%,40.1% and 37.4% in HDG,LC and HCC group,respectively,which showed no statistically significant difference(2χ= 0.547,P=0.05);there was a significant difference in BCP A1762T mutation rate between HDN and LC group(68.4% versus 36.3%,P=0.038).Compared with LC group,HCC group had a higher mutation detection rate of 73.0% on G1764A,with significant difference(P=0.0411).Mutation positive BCP gene was associated with significantly higher HBV DNA loads in LC and HDN group,compared with mutation negative BCP(2.18×107 versus 1.18×104,P=0.0451 in LC group,1.2×106 versus 2.95×103,P=0.0412 in HDN group,respectively);in addition,mutation positive BCP gene showed statistically higher HBeAg levels than mutation negative BCP gene in HDN and HCC group(750.00IU/ml versus 416.13 IU/ml,P=0.0451 in HDN group,1300.00IU/ml versus 927.60 IU/ml,P=0.0073 in HCC group,respectively).Conclusion BCP mutation was associated with clinical progression of hepatitis B nodular amyloidosis and played a vital role in malignant transformation of hepatitis B nodules to liver cancer.However,pre-C mutation was not involved in this process.Virus mutations and HBeAg quantification could predict the progress of chronic hepatitis B nodular change and have important clinical value.更多还原