摘要
目的探讨α-酮戊二酸/铁依赖性双加氧酶同源蛋白1(ALKBH1)基因低甲基化在肝细胞癌发病机制及病情评估中的临床价值。方法肝癌细胞(SMMC-7721、Bel-7402、MHCC97、HepG2及Hep3B)和正常肝细胞(HL-7702)购自美国典型培养物保存中心。选择2016年6月至2018年6月90例肝细胞癌患者为研究对象。采用重亚硫酸盐基因组测序聚合酶链反应(BSP)法检测ALKBH1基因甲基化并定量,采用甲基化特异性聚合酶链反应(MSP)法检测ALKBH1基因甲基化。统计并比较肝细胞癌患者肿瘤组织中ALKBH1基因甲基化和未甲基化患者平均疾病进展时间及3年生存率差异。计量资料比较行t检验,计数资料采用χ^(2)检验。结果本组肝细胞癌患者肿瘤组织ALKBH1基因甲基化定量显著低于癌旁正常组织[甲基化定量(15.6±1.7)%比(57.8±3.5)%],差异有统计学意义(χ^(2)=10.245,P<0.05)。SMMC-7721、Bel-7402、MHCC97、HepG2及Hep3B肝癌细胞ALKBH1基因甲基化定量显著低于HL-7702正常肝细胞[甲基化定量(25.1±2.1)%、(19.5±1.8)%、(16.7±1.3)%、(23.6±1.9)%、(20.3±1.5)%比(62.1±4.2)%],差异有统计学意义(χ^(2)=16.133,P<0.05)。本组肝细胞癌患者肿瘤组织ALKBH1基因甲基化率显著低于癌旁正常组织(25.6%比63.3%),差异有统计学意义(χ^(2)=36.897,P<0.05)。肝细胞癌患者肿瘤组织ALKBH1基因甲基化率在肿瘤最大径、病灶数量、分化程度和临床分期方面存在显著差异,肿瘤最大径≥5 cm、病灶数量多发、低中分化和Ⅲ期期肝细胞癌患者肿瘤组织ALKBH1基因甲基化率显著低于肿瘤最大径<5 cm、病灶数量单发、高分化和Ⅰ期+Ⅱ期肝癌患者(甲基化率分别21.2%比38.6%、5.0%比31.4%、12.8%比35.3%、4.0%比31.9%),差异有统计学意义(χ^(2)=4.328、4.406、4.745、4.881,均P<0.05)。肿瘤组织ALKBH1基因未甲基化患者平均疾病进展时间为(23.7±2.1)个月,3年生存率为52.4%,ALKBH1基因甲基化患者平均疾病进展时间为(29.5±2.7)
Objective To study the value ofα-ketoglutarate/iron-dependent dioxygenases homolog 1(ALKBH1)gene hypomethylation in the pathogenesis and disease assessment of hepatocellular carcinoma(HCC).Methods Hepatoma cells(SMMC-7721,Bel-7402,MHCC97,HepG2 and Hep3B)and normal hepatocytes(HL-7702)were purchased from the American Type Culture Collection.Patients with HCC were selected as the research subjects.The methylation of ALKBH1 gene was detected and quantified by bisulfite sequencing polymerase chain reaction(BSP)and methylation specific polymerase chain reaction(MSP)method.The difference in mean time to disease progression and 3-year survival rate between ALKBH1 methylated and unmethylated patients with HCC was calculated and compared.The measurement data were compared by t test,and the enumeration data were compared byχ^(2)test.Results The methylation quantification of ALKBH1 gene in tumor tissue was significantly lower than that in adjacent normal tissue[(15.6±1.7)%vs.(57.8±3.5)%],and the difference was statistically significant(χ^(2)=10.245,P<0.05).The methylation quantification of ALKBH1 gene in SMMC-7721,Bel-7402,MHCC97,HepG2 and Hep3B hepatoma cells were significantly lower than that in HL-7702 normal hepatocytes[(25.1±2.1)%,(19.5±1.8)%,(16.7±1.3)%,(23.6±1.9)%,(20.3±1.5)%vs.(62.1±4.2)%,χ^(2)=16.133,P<0.05].The methylation rate of ALKBH1 gene in the tumor tissue was significantly lower than adjacent normal tissue(25.6%vs.63.3%,χ^(2)=36.897,P<0.05).There were significant differences in the ALKBH1 gene methylation rate in terms of the maximum tumor diameter,the number of lesions,the degree of differentiation and the clinical stage(21.2%vs.38.6%,5.0%vs.31.4%,12.8%vs.35.3%,4.0%vs.31.9%,χ^(2)=4.328,4.406,4.745,4.881,all P<0.05).The mean time to disease progression and three-year survival rate of ALKBH1 unmethylated patients were(23.7±2.1)months and 52.4%respectively,and those of ALKBH1 methylated patients were(29.5±2.7)months and 78.8%respectively.The mean time to disease progression and three-year surv
作者
杨东昌
许珍
刘涛
赵守业
张磊
金秀
Yang Dongchang;Xu Zhen;Liu Tao;Zhao Shouye;Zhang Lei;Jin Xiu(Department of Hepatobiliary Surgery,Affiliated Hospital of Jining Medical College,Jining 272029,China;Department of Gastroenterology,Affiliated Hospital of Jining Medical College,Jining 272029,China;Department of Pathology,Affiliated Hospital of Jining Medical College,Jining 272029,China)
出处
《中华实验外科杂志》
CAS
北大核心
2022年第3期544-547,共4页
Chinese Journal of Experimental Surgery
基金
济宁市重点研发计划项目(2021YXNS065)。
关键词
肝细胞癌
甲基化
发病机制
Hepatocellular carcinoma
Methylation
Pathogenesis
