慢性乙型肝炎肝硬化结节样变与病毒变异的关系

Retrospective study on hepatic dysplastic nodules from hepatitis B virus(HBV)-related cirrhosis and HBV mutation

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。

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 G1896A was 52.3%,40.1 % and 37.4% in HDG,LC and HCC group, respectively,which showed no statistically significant difference （χ2 = 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 （750.00 IU/ml versus 416.13 IU/ml,P =0.0451 in HDN group,1300.00 IU/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.