A激酶锚定蛋白12基因甲基化与原发性肝癌复发的关系研究  

Association between A-kinase anchor proteins 12 methylation and recurrence of hepatocellular carcinoma

作　　者：刘兆君[1] 刘伟[2] 崔成华[1] 谷连坤[1] 邢宝才[2] 邓大君[1] 

机构地区：[1]北京大学肿瘤医院暨北京市肿瘤防治研究所,恶性肿瘤发病机制及转化研究教育部重点实验室,病因学研究室,100142 [2]北京大学肿瘤医院肝胆胰外一科,100142

出　　处：《中华预防医学杂志》2017年第9期848-852,共5页Chinese Journal of Preventive Medicine

基　　金：北京市自然科学基金（7133230）

摘　　要：目的 探讨A激酶锚定蛋白12（AKAP12）甲基化与原发性肝癌复发的关系.方法 于2003—2009年,以在北京大学肿瘤医院肝胆外科接受手术治疗的原发性肝癌患者作为研究对象,对其进行≥3年的随访.纳入标准：肝细胞癌患者,经鉴定切缘组织无肿瘤细胞存在;手术时无淋巴结转移及远处转移;有完整的临床资料,共142例.截至2014年5月随访结束,共有75例研究对象复发,其中71例死亡,无失访.从冰冻手术标本中获取研究对象的肿瘤组织及切缘组织,采用酚-氯仿法提取DNA并修饰后进行PCR扩增,对PCR产物进行克隆并测序分析,应用变性高效液相色谱检测研究对象肿瘤组织和肿瘤旁切缘正常组织AKAP12基因的甲基化状态.采用Log-rank检验对研究对象生存时间进行单因素分析;采用多因素Cox比例风险模型分析研究对象生存时间差异的影响因素.结果 142例研究对象中,男性125例（88.0％）,年龄为34～76岁,中位年龄是52.5岁.肿瘤组织中AKAP12甲基化阳性率（54.9％,78/142）高于切缘组织中AKAP12甲基化阳性率（10.2％,6/59）.与AKAP12甲基化阴性者相比,AKAP12甲基化阳性者原发性肝癌复发风险较低HR=0.62,95％CI：0.39～0.99;与肿瘤直径≤5 cm者相比,肿瘤直径〉5 cm者原发性肝癌复发风险较高HR=1.53,95％CI：1.00～2.50;与无门脉侵犯者相比,有门脉侵犯者原发性肝癌复发风险较高HR=4.53,95％CI：2.69～7.64.与无门脉侵犯者相比,有门脉侵犯者死亡风险较高HR=2.98,95％CI：1.73～4.98.结论 AKAP12甲基化与原发性肝癌的复发及无疾病生存时间相关.Objective To study the association between the AKAP12 promoter methylation and recurrence of hepatocellular carcinoma. Methods A total of 142 primary liver cancer patients underwent surgery in department of Hepatobiliary surgery in Peking University Cancer Hospital from 2003 to 2009 were selected as subjects in the survey; with the inclusion criteria as hepatocellular carcinoma, no cancer cells were observed in the surgical margin（SM） samples. All patients had neither lymph nor distant metastasis at the time of surgery, and receiving complete follow-up data for at least 3 years. By the end of May 2014, a total of 75 patients had relapsed of whom 71 died and there were no lost. All samples were acquired from the frozen surgical tissues. Genomic DNA was extracted using phenol/chloroform method and performed bisulfite modification following with polymerase chain reaction （PCR）. AKAP12 methylation in hepatoma and the corresponding SM samples from 142 patients was determined by denature high-performance liquid chromatography （DHPLC） and bisulfite clone sequencing. Kaplan-Meier and Cox proportion hazard regression model were used to identify the factors related to the survival time. Results In 142 cases, 125 patients （88.0%） were male and 17 （12.0%） cases were female. The median age was 52.5 years, ranging from 34 years to 76 years. AKAP12 methylation-positive rate was significantly higher in hepatomas than SMs （54.9%vs. 10.2%, P＆lt;0.001）. Patients with AKAP12 methylation-positive had less riskof the recurrence （HR=0.62, 95%CI：0.39-0.99）; with tumor diameter more than 5 cm （HR=1.53, 95%CI：1.00-2.50）,portal vein invasion（HR=4.53, 95% CI：2.69-7.64） increased the recurrence risk. Moreover, portal vein invasion had a higher risk of death （HR=2.98, 95% CI： 1.73-4.98）. Conclusion There was significant association between AKAP12 DNA methylation and low risk of recurrence and long progression-free survival of hepatocellular carcinoma patients.

关 键 词：肝肿瘤 DNA甲基化 复发 AKAPl2 

分 类 号：R735.7[医药卫生—肿瘤]