终末肝病患者血清和肝组织中乙型肝炎病毒转录体的对照研究

目的分析HBV转录体与终末肝癌肝硬化的关系。方法用血清核酸提取试剂盒及TRIzol从血清及肝组织中提取核酸,PCR及RT-PCR扩增HBV XDNAf、RNA和trRNA,琼脂糖电泳显示结果并进行分析。结果HBV DNA和转录体在HBsAg( )血清中的检出率显著高于HBsAg(-)中的检出率(P<0.01);fRNA在HBsAg( )/HBeAg( )血清中的检出率明显高于HBeAg(-)中的检出率(P<0.01)。trRNA在HBsAg(-)的隐匿性感染患者血清中有高检出率44.0%,且同一患者血清和组织中同时检出的一致率高。结论HBV转录体与终末期肝病的发生有密切的关系。trRNA具备作为诊断HBV感染新的血清学标志物的潜在可能。     

江苏省启东市乙型肝炎病毒基因型与肝癌及核心启动子突变的关系

目的 对江苏省启东市肝癌高发区的HBV流行株进行基因型别鉴定,并分析其与肝癌的相关性. 方法自启东市肝癌高发区的182份血清标本中抽提HBV DNA,以PCR方法扩增HBV全基因或X基因,测序后与基因库中已知基因型的参照序列共同构建基因进化树,根据序列在进化树中的位置进行型别鉴定.率的比较采用χ 2检验或Fisher精确概率法. 结果对包含24例HBV携带者、11例慢性乙型肝炎患者及13例肝癌患者在内的48例患者的血清HBV全基因测序结果显示,44例(91.7%)为C2基因型,4例(8.3%)为B2基因型,未见A、D、E、F、G、H等型别,也未见B/C重组病毒.以X基因测序方法对182例患者血清标本的鉴定结果显示,C2和B2型分别占92.9%(169例)和5.5%(10例),另有1.6%(3例)为B2/C2混合感染.启东地区C基因型的感染率明显高于邻近上海地区的78.9%(χ2=12.252,P<0.01).在81例肝癌和77例肝炎患者中,B、C基因型HBV的分布差异并无统计学意义(P>0.05).核心启动子区T1762/A1764联合突变在C基因型中的发生率为70.3%,明显高于B基因型中的30.8%(P<0.05);T1766和A1768突变则仅见于C基因型病毒中. 结论启东地区HBV感染以C2基因型为主,但其与肝癌的发生未见明显相关性.C基因型中较易出现核心启动子突变.     

乙型肝炎病毒基因变异、p16蛋白失活与肝癌的相关性研究

目的探讨乙型肝炎病毒（HBV）基因变异及肿瘤抑制基因p16蛋白失活与肝癌发生的关系。方法用基因芯片和核苷酸序列分析技术对114例HBV感染者（观察组）血清和43份肝组织标本中HBV DNA（前C区1814、1896、BCP区1762、1764位）进行分析,用流式细胞荧光染色法检测观察组和20例正常献血者（正常对照组）外周血白细胞中p16蛋白表达。结果观察组慢性重症乙型肝炎、肝硬化、肝癌患者血清HBV基因总突变率显著高于慢性乙型肝炎患者（P〈0．01）,肝硬化、肝癌肝组织HBV基因总突变率显著高于正常组织;正常对照组外周血自细胞中p16蛋白阳性率显著低于慢性乙型肝炎、肝硬化、肝癌患者,肝硬化、肝癌患者阳性率显著高于慢性乙型肝炎患者。结论HBV变异可能是肝癌发生的始动因子,而p16蛋白失活是肝癌发生的重要因素。     

慢性乙型肝炎病毒感染患者血清及肝组织中抗幽门螺杆菌抗体及特异性基因分析

目的 对慢性乙型肝炎病毒(HBV)感染患者血清抗幽门螺杆菌(Hp)IgG(抗Hp-IgG)阳性率进行流行病学调查,同时对患者肝组织进行Hp特异性基因检测,探讨Hp在肝病发生、发展中的作用.方法 病例对照研究中共纳入502例HBV感染患者和性别、年龄相匹配的429名健康对照者.应用酶联免疫吸附(ELISA)法进行血清抗Hp-IgG检测.同时用针对螺杆菌菌属特异性16S rRNA基因的通用引物对其中56例肝穿刺活检组织进行基因扩增,并对该基因阳性者进一步应用Hp cagA、vacA和glmM基因特异引物进行扩增.结果 HBV感染患者血清抗Hp-IgG阳性率为63.9%,显著高于健康对照者(43.4%,P<0.05),其中肝癌组的阳性率最高(29/36,80.6%),其次为肝硬化组(64/83,77.1%),两组均显著高于慢性乙型肝炎组(228/383,59.5%,P<0.01).56例行肝穿刺活检患者中,35例肝组织中发现螺杆菌菌属特异性16S rRNA基因,其中肝硬化组17例,肝癌组7例,慢性乙型肝炎组11例.进一步的扩增结果 证实35例中21例为Hp DNA.结论 HBV感染患者血清抗Hp-IgG阳性率显著高于健康对照者.HBV感染患者肝组织中除存在Hp DNA外,可能还存在其他螺杆菌DNA.螺杆菌在慢性乙型肝炎向肝硬化和肝癌的发展过程中可能发挥致病作用.     

乙型肝炎患者重叠感染HCV及HDV临床意义探讨

本文对305例乙型肝炎患者的血清标本回顾性地检测了HCV及HDV感染标志,结果有48.2％的乙肝患者重叠感染了HCV和/或HDV,其中双重感染率:HBV/HCV 15.4％,HBV/HDV 21.0％;三重感染率:HBV/HCV/HDV同时感染占11.8％,急性盱炎(AH)、厄症状表面抗原携带者(ASC),慢迁肝(CPH).慢活肝(CAH)、肝硬化(CIR)、重型肝炎(SH)及原发性肝癌(PHC)患者的抗-HCV检出率分别为:6.7％、6.7％、22.5％.29.7％、36.4％、38.5％、33.3％:其DHV的检出率分别为:10.0％、13.3％、25.0％、35.9％、41.1％、53.8％、42.9％。肝病越严重,其HCV及HDV的感染率就越高,提示HBV重叠感染HCV及HDV时可以促使肝脏病变加重。合并HCV及HDV感染者的HBV复制率较低。     

乙型肝炎病毒核心启动子缺失突变的初步观察

目的研究乙型肝炎病毒(HBV)e抗原阴性患者HBV核心基因启动子变异方式。方法自5例慢性HBV感染者血清中分别提取HBVDNA,应用多重PCR法鉴定其基因型。PCR扩增X基因序列,克隆入pMD19T载体,共挑选23个克隆测序,与已知HBV基因相应序列比较该患者体内HBV基因变异程度。结果 5例患者中3例基因型分别为B、C和B/C混合型,2例患者通过现行多引物方法无法分型。16个(16/23,69.6%)克隆在X基因下游出现大段缺失突变,其中15例缺失长度为234bp,1例为245bp。缺失突变区域包括C基因启动子区,并导致前-C区起始密码子ATG编码缺失,这种缺失突变在5例患者中均被检出。结论 CP/HBeAg起始密码子缺失突变可能是HBeAg阴性慢性乙型肝炎的一种变异模式。     

慢性乙型肝炎患者HBe系统与HBV DNA前C区基因突变的分析

目的研究HBV DNA前C区基因突变与HBe系统的关系.方法对88例慢性乙型肝炎患者进行HBVDNA基因检测(PCR法)、分子克隆及序列分析.结果88例患者中,血清HBV DNA阴性22例,阳性66例.从阳性患者中检出野生株感染22例,占33.3%;突变株感染44例,占66.7%.44例突变株感染者中38例属1898位点及/或1901位点突变,占86.4%;其他形式突变6例,占13.6%;66例血清HBV DNA阳性中,CHB 49例,LC 17例,突变株感染检出率分别为69.3%、58.8%,两者经统计学处理无显著性差异(P＞0.05).在HBe系统中,HBV DNA阳性患者属HBeAg(+)者44例,其中突变株检出率为75.0%(33/44),属抗HBe(+)者20例,其中突变株检出率为50.0%(10/20),两者经统计学处理有显著性差异(P＜0.05).结论①在HBe系统中,HBeAg(+)患者的突变株检出率高于抗HBe(+)患者(P＜0.05),提示其位点突变株在HBeAg(+)中占优势,与以往文献报道中关于1896位点突变株主要存在于抗-HBe(+)患者中的结论不尽相同.因此,应考虑HBeAg(+)患者也存在病毒突变的可能.②HBV慢性感染过程中,CHB患者与LC患者的突变株感染检出率无显著性差异(P＞0.05),表明在1898及/或1901位点的突变株感染与肝脏损害程度无平行关系.     

多种肝脏疾病中隐匿性HBV感染检出率及其S基因MHR免疫逃逸相关突变特点分析

目的分析多种肝脏疾病中隐匿性HBV感染(occult HBV infection,OBI)检出率,并探讨OBI患者HBV S基因主要亲水区(major hydrophilic region,MHR)免疫逃逸相关突变特点。方法回顾性分析2005年1月—2017年12月就诊于中国人民解放军总医院第五医学中心的91037例HBV感染住院患者临床资料,筛选出OBI患者并扩增其HBV S基因序列,分析其HBV S基因MHR免疫逃逸相关突变特点。结果91037例住院患者中OBI总检出率为0.53%(487/91037),急性乙型肝炎患者中OBI检出率最高(9.26%,130/1404),肝硬化患者中OBI检出率最低(0.26%,78/29921)。62例OBI患者组与124例非OBI患者组相比,OBI患者组MHR免疫逃逸相关突变总体检出率显著高于非OBI患者组(59.68%vs.35.48%;P<0.05);OBI患者组MHR多个免疫逃逸相关突变的联合检出率显著高于非OBI患者组(43.55%vs.22.58%;P<0.05);其中,sT118K、sK122R和sV168A 3种单点突变的检出率显著高于非OBI患者组。结论本研究显示临床HBV感染患者中有较高的OBI检出率,而且不同肝脏疾病中OBI检出率不同。此外,HBV S基因MHR的免疫逃逸相关突变与临床实践中OBI的发生密切相关。     

厦门市乙型肝炎患者病毒基因型的分析

目的:采用多对型特异性引物,通过巢式PCR法检测厦门市乙型肝炎患者血清中乙型肝炎病毒(HBV)基因型的分布情况.方法:收集250例HBV感染患者血清,提取血清中HBV DNA作为模板,设计HBV前S1基因和S基因中区域内设计出10条内外引物,并将其中8条型特异性内引物分成A,B两组,分别扩增A,B,C和D,E,F型HBV,然后将第2轮PCR产物以用30g/L琼脂糖进行电泳,根据PCR产物电泳显示的产物长度判定HBV基因型,以了解厦门HBV基因型分布情况.结果:共120例确定了HBV基因型.患者群中慢性乙型肝炎90例,占75.0%,急性乙型肝炎、肝炎肝硬化、原发性肝癌分别占5.8%(7/120)、6.7%(8/120)和12.5%(15/120).分型结果:B型58例(48.3%)、C型30例(25.0%),B/C混合型32例(26.7%).HBeAg阳性患者中B基因型占63.8%,B/C型混合感染21.9%;抗-HBe阳性患者中以B/C型混合感染68.8%,B型25.9%,HBeAg阳性组与抗-HBe组之间比较发现B型和B/C混合型之间(P<0.05).结论:厦门乙型肝炎患者HBV基因型以B型为主,B/C混合感染是一个值得重视的问题.     

合并乙型肝炎病毒感染对丙型肝炎病毒核心抗原检测的影响

目的 探讨合并HBV感染对慢性HCV感染者血清丙型肝炎病毒核心抗原(HCVcAg)检出情况的影响. 方法 收集2005年12月-2009年10月慢性丙型肝炎患者和HBV/HCV合并感染者资料,检测血清HCVcAg和HCV RNA,对后者血清进行HBV DNA、HBeAg检测,分析HCVcAg检出率与HBeAg、HBV DNA定量检测的关系.用独立两组多分类的X2检验方法进行统计学分析. 结果 共收集88例慢性丙型肝炎患者和62例HBV/HCV合并感染者资料,血清HCVcAg的检出率分别为72.7％(64/88)和38.7％ (24/62),两者比较,x2= 17.358,P＜0.01,差异有统计学意义.HCV RNA检出率分别为81.8％ (72/88)和53.2％ (33/62),两者比较,x2=20.110,P＜0.01,差异有统计学意义.62例HBV/HCV合并感染者血清中,HBeAg阳性和HBeAg阴性感染者HCVcAg检出率分别为28.6％ (12/42)和60.0％ (12/20),两者比较,x2=5.641,P=0.011,差异有统计学意义.HCV RNA阳性率分别为42.9％ (18/42)和80.0％ (16/20),两者比较,X2=7.547,P＜ 0.01,差异有统计学意义.HBV DNA阳性和阴性时HCVcAg检出率分别为39.1％ (18/46)和37.5％ (6/16),两者比较,P＞0.05,差异无统计学意义.与单纯HCV感染者血清HCVcAg检出率72.7％ (64/88)比较,HBeAg阴性合并感染者为60.0％ (12/20),x2=1.266,P=0.261,差异无统计学意义;HBV DNA阴性合并感染者为37.5％ (6/16),x2=7.635,P＜0.01,差异有统计学意义.结论 HBV/HCV合并感染时HCVcAg检出率较低,可能是由于HBeAg抑制HCV的复制,从而减少HCVcAg的表达所致.     

Overexpression of HBxAg in hepatocellular carcinoma and its relationship with Fas／FasL system

AIM: To study the expression and serum level of HBxAg,Fas and FasL in tissues of HCC patients, and to assess the relationship between HBxAg and Fas/FasL system.METHODS: Tissues from 50 patients with HCC were tested for the expression of HBxAg, Fas and FasL by S-P immunohistochemistry. Serum levels of sFas/sFasL and HBsAg/HBeAg were measured by ELISA assay. HBV X gene was detected by PCR in serum and confirmed by automatic sequencing. Fifty cases of liver cirrhosis and 30 normal controls were involved in serum analysis.RESULTS: The expression of HBxAg, Fas and FasL in carcinoma tissues was 96 %, 84 % and 98 %, respectively.Staining of HBxAg, Fas and FasL was observed predominately in cytoplasms, no significant difference was found in intensity between HBxAg, Fas and FasL (P＞0.05). HBxAg, Fas and FasL might express in the same area of carcinoma tissues and this co-expression could be found in most patients with HCC. The mean levels of sFas in serum from HCC, cirrhosis and normal controls were 762.29±391.56 μg@ L-1 835.36±407.33 μg@L-1 and 238.27±135.29 μg@L-1. The mean levels of sFasL in serum from HCC, cirrhosis and normal controls were 156.36±9.61iμg@ L-1, 173.63±18.74 μg@L-1 and 121.96±7.83 μg@ L-1.Statistical analysis showed that both sFas and sFasL in HCC and cirrhosis patients were significantly higher than those in normal controls (P＜0.01). Serum HBV X gene was found in 32 % of HCC patients and ,46 % of cirrhotic patients.There was no significant relationship between serum level of sFas/sFasL and serum X gene detection (P＞0.05). Eight percent of HCC patients with negative HBsAg and HBeAg in serum might have X gene in serum and HBxAg expression in carcinoma tissues.CONCLUSION: Our data suggest that HBxAg and Fas/FasL system plays an important role in the development of human HCC. Expression of HBxAg can leads to expression of Fas/FasL system which and reverse apoptosis of hepatocellular carcinoma induced by FasL.     

Characterisation of hepatitis B virus X protein mutants in tumour and non-tumour liver cells using laser capture microdissection

BACKGROUND/AIMS: The analysis of hepatitis B virus (HBV) X protein genetic variability and is correlation with liver disease severity have only been addressed, so far, on whole liver extracts. We have studied, therefore, the HBV X protein (HBx) gene sequence in morphologically well-characterised tumour and non-tumour liver cells from patients with HBV-related hepatocellular carcinoma. METHODS: Using laser capture microdissection (LCM), we picked up six to eight groups of tumour and non-tumour hepatocytes in serial frozen sections from six patients. After global DNA preamplification followed by HBx-specific polymerase chain reaction, the HBx gene was sequenced in each group of microdissected cells. We also validated the quantification of HBV-DNA in microdissected hepatocytes using HBV Amplicor. RESULTS: Heterogeneous mutations in HBx gene were found in distinct cirrhotic nodules and tumour areas from the same patient. Mutations at aa 127, 130 and 131 were frequently detected but there was no distinct point mutation profile between tumour and non-tumour samples. In contrast, deletions in HBx gene, which were found in five/six patients, were more frequent in tumour-derived sequences (6/18) than in non-tumour-derived sequences (1/20). CONCLUSIONS: We have shown that LCM provides a direct insight of intrahepatic HBV infection. Using this technique, we demonstrated the persistence of distinct HBx encoding sequences in clonally expanding cells, thus supporting the hypothesis that HBx deletions may be implicated in liver carcinogenesis.     

Factors predicting occurrence and prognosis of hepatitis-B-virus-related hepatocellular carcinoma

Primary liver cancer is an important cause of cancer death,and hepatocellular carcinoma(HCC) accounts for 70%-85% of total liver cancer worldwide.Chronic hepatitis B virus(HBV) infection contributes to > 75% of HCC cases.High serum viral load is the most reliable indicator of viral replication in predicting development of HCC.HBV genotype C is closely associated with HCC in cirrhotic patients aged > 50 years,whereas genotype B is associated with development of HCC in non-cirrhotic young patients and postope...     

High prevalence of 1762(T) 1764(A) mutations in the basic core promoter of hepatitis B virus isolated from black Africans with hepatocellular carcinoma compared with asymptomatic carriers

The purpose of this study was to identify mutations in the basic core promoter and enhancer II region of the hepatitis B virus (HBV) that might cause the HBV e antigen (HBeAg)-negative phenotype and contribute to hepatocarcinogenesis in black African carriers of the virus. The basic core promoter/enhancer II overlaps with the X gene. HBV DNA from serum of 47 asymptomatic carriers and 50 patients with hepatocellular carcinoma and from 28 tumor and 10 nontumor liver tissues was amplified and sequenced directly. That part of the enhancer II region not overlapping the basic core promoter was completely conserved in all samples. Missense mutations at nucleotides 1809 and 1812 in the basic core promoter were found in 80% of all sequences and may represent wild-type sequence in Southern African isolates. Nucleotide and amino acid divergences were higher in the basic core promoter of hepatocellular carcinoma patients when compared with asymptomatic carriers (P <.0001). This applied particularly to the paired 1762 adenine to thymine (1762(T)) and 1764 guanine to adenine (1764(A)) missense mutations, the prevalence of which was 66% in patients with hepatocellular carcinoma compared with 11% in asymptomatic carriers (P <.0001). There was no association between the presence of 1762(T) 1764(A) and HBeAg negativity, although these mutations suppressed HBeAg titers in HBeAg-positive patients. Suppression of HBeAg expression as well as alteration of the amino acid sequence of the X protein may play a role in hepatocarcinogenesis.     

Expression of nuclear factor-kappa B in hepatocellular carcinoma and its relation with the X protein of hepatitis B virus

AIM: In this study we investigated the relationship of the X protein of HBV and nuclear factor-kappa B (NF-kappa B) and the expression of NF-kappa B in human hepatocellular carcinoma tissues. METHODS: Immunohistochemistry SP method was used to detect the expression of NF-kappa B and the X protein of HBV in human hepatocellular carcinoma tissues of 52 cases. Gene transfection mediated by lipofectamine was used to transfect the eukaryotic expression vector pCDNA3.1-HBX of HBV x gene into human hepatocellular carcinoma cell line HCC-9204 and NF-kappa B was detected. RESULTS: NF kappa B was widely expressed in human hepatocellular carcinoma tissues in a total of 52 cases and its expression was related to the X protein of HBV. NF-kappa B was localized both in the cytoplasm and the nuclei of hepatocellular carcinoma cells in 11 cases which were positive for the X protein of HBV while in 41 cases negative for the X protein of HBV, NF-kappa B was only localized in the cytoplasm of hepatocellular carcinoma cells but translocated to the nuclei of hepatocellular carcinoma cells after the eukaryotic expression vector pCDNA3.1-HBX was transfected into HCC-9204 cells. CONCLUSION: This study strongly suggests that the nuclear factor NF-kappa B is widely expressed in hepatocellular carcinoma tissues in different styles according to the expression of the X protein of HBV. NF-kappa B is abnormally activated in hepatocellular carcinoma, which is probably related to the X protein of HBV. The X protein of HBV can activate NF-kappa B to translocate into nuclei of hepatocellular carcinoma cells.     

Clinical usefulness of serum matrix metalloproteinase-2 concentration in patients with chronic viral liver disease

BACKGROUND/AIMS: The production of matrix metalloproteinase (MMP)-2 was reported to be increased in chronically diseased livers. Our aims in the present study were to elucidate the clinical usefulness of the serum MMP-2 concentration in chronic viral liver disease. METHODS: We measured serum MMP-2 concentrations with a sandwich enzyme immunoassay in 62 patients with chronic hepatitis, 35 patients with liver cirrhosis, 55 patients with hepatocellular carcinoma and 24 healthy individuals. The assay detects proMMP-2 and proMMP-2 complexed with tissue inhibitor of metalloproteinase-2, but not active forms of MMP-2. The liver MMP-2 content was also measured in autopsied cirrhotic and non-cirrhotic livers. Gelatin zymography and gel filtration chromatography were carried out using the serum. RESULTS: The serum MMP-2 concentration was significantly increased in the liver cirrhosis and hepatocellular carcinoma patients, but not in the patients with chronic hepatitis. There was no significant difference in the serum MMP-2 level between the liver cirrhosis and hepatocellular carcinoma groups. In the patients with chronic viral liver disease, serum MMP-2 concentration showed the best correlation with the degree of liver fibrosis and with serum hyaluronate level. The zymography of serum showed the majority of MMP-2 in serum exists as a proMMP-2. The chromatography of serum revealed a single peak at the position of about 90 kDa corresponding to an MMP-2 complexed with tissue inhibitor of metalloproteinases-2. The liver MMP-2 content was markedly increased in the cirrhotic livers compared with the non-cirrhotic livers, and was positively correlated with the liver collagen content. When investigating the utility of the serum MMP-2 test for differentiating liver cirrhosis from chronic hepatitis, the utility of serum MMP-2 was equal to that of serum hyaluronate, which is known as the best current test for diagnosing liver cirrhosis. CONCLUSIONS: The serum MMP-2 concentration reflects mainly the amount of proMMP-2 complexed with tissue inhibitor of metalloproteinase-2. The serum MMP-2 level was markedly increased in cirrhotic patients, and may be explained by an overproduction in the cirrhotic liver. In the clinical state, the measurement of serum MMP-2 was as useful a test for diagnosing liver cirrhosis as is the serum hyaluronate level.     

Mutation of p53 gene in regenerative nodules in cirrhotic liver

BACKGROUND/AIMS: Mutations of p53 gene have been detected in precancerous stages of several cancers, and the possible role in multistep carcinogenesis is suggested. The aim of this study was to examine the mutation profile of p53 gene in regenerative nodules in cirrhotic livers.METHODS: Ninety eight tissue specimens of regenerative nodules obtained from 15 cases of cirrhosis were used for analysis. Twenty cases of chronic hepatitis and two cases of fatty liver were used as controls. DNA was extracted from each of manually demarcated regenerative nodules, and nucleotide sequence analysis was performed on p53 gene exon 5.RESULTS: Direct sequencing detected p53 mutations in seven of 98 DNA samples (7.1%) from regenerative nodules in six cases of cirrhosis. Subcloning analysis revealed that mutation sites differed in each subclone and the incidences of the mutation varied from 7.7 to 58.8% depending on individual nodules. The mutation was not detected in any of chronic hepatitis and fatty liver. There were inconsistent p53 sequence with regenerative nodules and accompanied hepatocellular carcinomas in six cases.CONCLUSIONS: Mutations of p53 gene were frequently found in cirrhotic livers compared with livers of patients with chronic hepatitis (P<0.01), suggesting that p53 mutations at the stage of cirrhosis may be a causative factor that may potentially lead to hepatocellular carcinoma.     

肝细胞癌和肝硬变组织中突变型P53蛋白的表达及与HBV DNA的关系

应用LSAB免疫组化技术,检测肝内突变型P53蛋白的表达;用地高辛探针原位杂交检测HBV DNA。结果显示,31例肝细胞癌(HCC)及其癌旁组织中突变型P53蛋白阳性率分别为45％和48％,而11例肝硬变中检出4例(36.4％)。HBV DNA阳性率分别为46％和86％。癌旁组织中突变型P53蛋白表达与HBV DNA存在相关关系(P<0.05)。提示P53基因突变与HBV慢性感染有关,可能是诱发肝癌的机制之一。癌旁组织和肝硬变组织中也存在P53蛋白异常表达,表明肝癌形成早期就发生了53基因的突变。     

青岛地区乙型肝炎病毒基本核心启动子区基因突变与乙型肝炎病毒所致原发性肝癌的相关性

目的通过分析山东省青岛地区HBV核心启动子基因序列的突变特征,探讨其与乙型肝炎相关原发性肝癌的相关性。方法收取慢性乙型肝炎患者和乙型肝炎相关原发性肝癌患者的血清标本各60例,然后从中提取HBV DNA,采用聚合酶链反应(PCR)扩增、纯化、克隆后测序,根据S基因区的编码序列确定患者的基因型和血清型;分别将HBV核心启动子区的各序列结果与GeneBank中的HBV标准株做对比,用DNAMAN软件对基因序列进行突变分析。采用SPSS17.0软件进行统计学分析。结果 120例标本,HBV株均为B或C基因型,以C基因型为主,其中,HBV组的C基因型所占比率为83.33%,HCC组为90.00%(χ2=0.65,P=0.42);血清型均为adw2或adrq+。HBV基因组核心启动子区常见的点突变为C1653T、T1753V、C1754T及A1762T/G1764A,发生率分别为42.73%、86.36%、71.82%、38.18%。与慢性乙型肝炎患者相比,肝癌患者中发生率比较高的突变位点为C1653T(78.85%,χ2=52.58,P<0.001)及A1762T/G1764A(73.08%,χ2=50.88,P<0.001)。结论山东青岛地区HBV基因组常见为B、C基因型,其中以C型为主;核心启动子区突变发生率高,其中,T1753V与HCC发生无关,C1653T、A1762T和G1764A位点无论单独或是联合突变均与HCC发生密切相关。