CAJ | 학술논문

目的探讨肝细胞癌(HCC)患者CDH1 、GSTP1和RASSF1A基因启动子区域异常甲基化以及淋巴细胞微核对肝细胞癌发生风险的影响.方法应用甲基化特异性PCR(MSP)技术对32例HCC患者肿瘤组织和癌旁组织的CDH1、GSTP1和RASSF1A基因启动子区域进行甲基化检测.未经治疗前抽取HCC患者外周血进行体内淋巴细胞微核试验,比较HCC自发微核率和健康人群对照组(n=50)的差异.结果 32例HCC组织中,CDH1、GSTP1和RASSF1A基因甲基化检出率分别是43.8％(14/32)、68.8％ (22/32)和43.8％ (14/32),均高于相应的癌旁组织[25％(8/32)、46.9％(15/32)、15.6％(5/32)].只有RASSF1A基因在HCC组织和癌旁组织中的甲基化检出率差异有统计学意义(43.8％比15.6％,P＜0.05).81.3％(26/32)的HCC和65.6％(21/32)的癌旁组织中有1个或1个以上基因发生了甲基化.3个基因的甲基化程度与患者年龄、性别、肿瘤型、分化程度等临床病理参数均无统计学关联.HCC组微核率与对照组差异有统计学意义(1.38‰比0.47‰,P＜0.05);同时HCC患者自发微核率随着病理分化变差而呈现逐渐增高的趋势,高中、中、中低分化HCC患者微核率分别为1.30‰、1.33‰和1.64％‰.结论 CDH1、GSTP1和RASSF1A基因启动子区域异常甲基化和染色体不稳定存在于HCC的发生发展中.基因启动子区域异常甲基化可能与HCC发生早期密切相关.甲基化检测对于HCC高危人群筛选以及HCC早期辅助诊断具备一定的应用价值.
목적 탐토간세포암(HCC)환자CDH1 、GSTP1화RASSF1A기인계동자구역이상갑기화이급림파세포미핵대간세포암발생풍험적영향.방법 응용갑기화특이성PCR(MSP)기술대32례HCC환자종류조직화암방조직적CDH1、GSTP1화RASSF1A기인계동자구역진행갑기화검측.미경치료전추취HCC환자외주혈진행체내림파세포미핵시험,비교HCC자발미핵솔화건강인군대조조(n=50)적차이.결과 32례HCC조직중,CDH1、GSTP1화RASSF1A기인갑기화검출솔분별시43.8％(14/32)、68.8％ (22/32)화43.8％ (14/32),균고우상응적암방조직[25％(8/32)、46.9％(15/32)、15.6％(5/32)].지유RASSF1A기인재HCC조직화암방조직중적갑기화검출솔차이유통계학의의(43.8％비15.6％,P＜0.05).81.3％(26/32)적HCC화65.6％(21/32)적암방조직중유1개혹1개이상기인발생료갑기화.3개기인적갑기화정도여환자년령、성별、종류형、분화정도등림상병리삼수균무통계학관련.HCC조미핵솔여대조조차이유통계학의의(1.38‰비0.47‰,P＜0.05);동시HCC환자자발미핵솔수착병리분화변차이정현축점증고적추세,고중、중、중저분화HCC환자미핵솔분별위1.30‰、1.33‰화1.64％‰.결론 CDH1、GSTP1화RASSF1A기인계동자구역이상갑기화화염색체불은정존재우HCC적발생발전중.기인계동자구역이상갑기화가능여HCC발생조기밀절상관.갑기화검측대우HCC고위인군사선이급HCC조기보조진단구비일정적응용개치.
Objective To explore the effects of aberrant methylation in the CDH1,GSTP1 and RASSF1A gene promoter CpG region and lymphocyte micronucleus on the pathogenesis of hepatocellular carcinoma(HCC).Methods The methylation-specific polymerase chain reaction(MSP)was employed to investigate the promoter methylation of CDH1,GSTP1 and RASSF1A genes in the tumor and adjacent tissues in 32 patients with HCC.The peripheral blood was sampled from patients with HCC for subsequent micronucleus test for lymphocytes,prior to the treatment.The differences of the spontaneous micronucleus rate in HCC patients and healthy population control group(n=50) were compared.Results Tumor tissues of patients with HCC yielded higher assay sensitivity for detection of methylation of CDH1,GSTP1 and RASSF1A genes[43.8％(14/32),68.8％(22/32) and 43.8％ (14/32)] than adjacent tissues [25％ (8/32),46.9％ (15/32) and 15.6％ (5/32)].Nevertheless,the between-group difference in methylation for various genes did not reach statistical significance with exception of RASSF1A gene(43.8％ vs 15.6％,P＜0.05).Methylation in one or more of the three genes was noted in tumor tissues[81.3％(26/32)] and adjacent tissues[65.6％(21/32)].Additionally,the status of methylation was not statistically associated with the age,sex,type of tumor and tumor differentiation.Patients with HCC,who showed a raised risk of automatic micronucleation with the advanced pathologic differentiation,were featured by a higher rate of micronucleation when compared with normal controls(1.38‰ vs 0.47‰,P＜0.05).The frequency of micronucleation was 1.30‰,1.33‰ and 1.64‰o in patients with moderate-to-high,moderate and low differentiation of HCC.Conclusions Genetic instability and promoter methylation of CDH1,GSTP1 and RASSF1A genes may play a vital role in the pathogenesis of HCC.Early stage HCC may be closely linked to aberrant promoter region methylation.Methylation detection is valuable in screening high-risk population and establishing adjunct early diagnosis of HCC.