不良结局聚集性乙型肝炎病毒感染家系基本核心启动子／前C区热点突变分析

Analysis of hot mutations of basic core promoter and PreC region in familial clustering of hepatitis B virus infection with unfavorable prognoses

目的了解HBV基本核心启动子（BCP）区及前C（PreC）区突变在不良结局家族聚集性HBV感染家系中不同疾病进展阶段的分布特点。方法共纳入39个不良结局家族聚集性HBV感染家系，利用PCR及直接测序法对慢性HBV携带者53例，CHB患者56例，肝硬化患者43例和肝细胞癌（HCC）患者28例进行基因分型及BCP／PreC序列分析。统计学处理采用独立样本t检验、χ2检验或Fisher确切概率法。结果在HCC患者中，A1762T／G1764A、C1766T、T1768A、A1846T、G1899A突变发生率分别为64．29％、25．00％、25．00％、28．57％和17．86％，显著高于慢性HBV携带者的37．74％、1．89％、0、7．55％和3．77％，差异有统计学意义（χ2值分别为5．186、8．885、14．503、4．859、4．603，均P〈0．05）；其中C1766T、T1768A突变在HCC患者中的发生率显著高于CHB患者的8．93％和5．36％，差异有统计学意义（χ2值分别为3．938、6．868，均P〈0．05）。A1762T／G1764A在C基因型中的突变发生率（59．12％）显著高于B基因型（16．13％），差异有统计学意义（χ2=13．118，P〈0．05）；A1752V、C1799G突变发生率在B基因型分别为90．32％和96。77％，显著高于C基因型的4、38％和1．46％，差异有统计学意义（7。值分别为110．409、142．825，均P〈0．05）。Logistic回归分析显示，与慢性HBV携带者比较，高龄、男性、ALT、AsT、HBVDNA〉1×10^4IU／mL、A1762T／G1764A、G1896A、C1766T、T1768A、G1899A、A1846T突变与HCC独立相关。与非HCC比较，高龄、男性、ALT、AST、G1896A、C1766T、T1768A、A1846T突变与HCC独立相关。结论A1762T／G1764A、C1766T、T1768A、A1846T、G1899A突变与不良结局家族聚集性HBV感染家系的疾病进展相关，主要与HCC进展相关；A1762T／G1764A、A1752V、C1799G突变在C基因到与B慕因型中存在苹异。

Objective To investigate the distribution of hot mutations of basic core promoter （BCP） and PreC region in different stages of familial clustering of hepatitis B virus （HBV） infection with unfavorable prognoses. Methods Thirty-nine families with clustering of infection with unfavorable prognoses were enrolled in this study. All samples including 53 chronic HBV carriers, 56 chronic hepatitis B （CHB） patients, 43 liver cirrhosis （LC） patients and 28 hepatocellular carcinoma （HCC） patients were analyzed for genotypes and BCP/PreC mutations with polymerase chain reaction （PCR） and direct sequencing. Independent-samples t test was used for continuous variables, while χ2 test or Fisher exact probability test was used for dichotomous variables. Results Proportions of A1762T/G1764A, C1766T, T1768A, A1846T and G1899A in HCC patients were 64.29%, 25.00%, 25.00%, 28.57% and 17.86%,respectively, which were significantly higher than those in chronic HBV carriers （37.74 %, 1.89%, 0, 7.55 and 3.77%, respectively; χ2= 5. 186, 8. 885, 14. 503, 4. 859 and 4. 603, respectively, all P〈0.05）. C1766T and T1768A were significantly associated with HCC compared with CHB which were 8. 93% and 5.36%, respectively （χ2 =3. 938 and 6. 868, respectively; both P〈0.05）. Mutation of A1762T/G1764A ingenotypeC （59.12%） was higher than genotype B （16. 13% χ2=13.118, P〈0.05）. Mutations of A1752V and C1799G in genotype B were 90.32% and 96.77%, respectively, which were higher than that in genotype C （4. 38% and 1. 46%, respectively; χ2 = 110. 409 and 142. 825, respectively; both P〈 0.05）. Logistic regression analysis showed that old age, male gender, abnormal alanine aminotransferase （ALT）, abnormal aspartate transaminase （AST）, HBV DNA〉 1 × 10^4 IU/mL, A1762T/G1764A, G1896A, C1766T, T1768A, G1899A and A1846T were independently associated with HCC compared with chronic HBV carriers. Old age, male gender, abnormal ALT, abnormal AST, G1896A, C1766T, T1768A and A1846T were independently associated with HCC compared with those without HCC. Conclusions A1762T/G1764A, C1766T, T1768A, A1846T and G1899A are associated with HCC in familial clustering of HBV infection with unfavorable prognoses. Distributions of A1762T/G1764A, A1752V and C1799G are different between genotype B and C.