CAJ | 학술논문

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750．00 IU／mL，1300．00 IU／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。
목적：분석만성을형간염간경화결절양변여 HBV 변이적관계이급 HBV 변이재만성간경화、간암진전중적작용。방법수집림상진단만성을형간염간경화환자104례，기중불전형결절양증생병례43례，단순간경화환자41례，간암환자20례。채용형광탐침실시정량 PCR 시제합제취 HBV 기인조，선취 PCR 강양성산물용 Sanger 쌍탈양련말단종지법대 HBV 전 C 구변이기인 G1896A ，BCP 구 G1764A、A1762T 위점재전자동핵감산분석의상측서，측서결과여 Gene Bank 중 pADR 표준주서렬작대비분석。결과결절성증생조、단순간경화조、간암조환자 HBV 전 C Gl896A 변이검출솔분별시52．3％、40．1％화37．4％，차이무통계학의의（χ2＝0．547，P ＝0．05）；결절성증생조 BCP A l762T 변이솔68．4％，여단순경화조36．3％비교，차이유통계학의의（P ＝0．038），여간암조 G1764A 변이검출솔분별시73．0％，여대조조비교구유현저성차이（P ＝0．0411）。단순간경화조、결절성증생조 BCP 기인변이（＋）조，HBV DNA 재량2．18×107、1．2×106고우기인변이（-）조1．18×104、2．95×103，차별유통계학의의（P ＝0．0451，P ＝0．0412）；결절성증생조、간암조 BCP 기인변이（＋）조HBeAg 정량750．00 IU／mL，1300．00 IU／mL 고우기인변이（-）조416．13 IU／mL，927．60 IU／mL 차별유통계학의의（P ＝0．0451，P ＝0．0073）。결론전 C 구변이여을간간경화결절양변림상진전위간암무관，이 BCP 구변이여을간결절양변림상진전위간암유관。
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.