摘要
目的:利用基于女性体细胞构成组织内X染色体失活嵌合性的磷酸甘油酸激酶和雄激素受体位点克隆性检测技术,探讨肝硬化组织中变异肝细胞结节(NAH)的克隆性.方法:女性乙型肝炎病毒(HBV)相关肝硬化标本石蜡切片,HE染色后通过显微切割获取肝细胞结节,提取基因组DNA,经甲基化敏感的HpaⅡ或HhaⅠ消化,巢式PCR扩增,凝胶电泳显示结果.应用肝细胞癌(HCC)和肝细胞腺瘤组织作为参照病变.结果:HCC5例和肝细胞腺瘤组织1例均为单克隆性.在4例可分析的肝硬化组织中共分离出普通再生结节6个和NAH29个,后者包括混合细胞性病变25个,透明细胞性(糖原储积)病变4个.伴有小细胞性改变(SCC)的5个混合细胞性NAH均显示X染色体失活嵌合性丢失,提示为肿瘤性病变;其中1个取自癌旁的病变与癌组织的灭活带型一致,证明与HCC具有相同的克隆起源;20个不伴SCC的混合细胞性NAH中,7个(35%)为单克隆性病变;检测的4个透明细胞性NAH和6个普通再生结节均未显示出X染色体失活嵌合性丢失,提示为多克隆性病变.结论:HBV相关的肝硬化组织中的部分NAH,尤其是伴有SCC的病变,是单克隆性的,已经属于肿瘤性病变;SCC是NAH病变进展过程中的较晚期改变,是一种癌前病变.
AIM: To elucidate the clonality status of the preneoplastic nodules of altered hepatocytes (NAHs) in cirrhotic liver by clonality assays at the phosphoglycerate kinase and an-drogen receptor loci based on X chromosomal inactivation mosaicism in female somatic tissues. METHODS: Female liver tissues with hepatitis B virus (HBV)-associated cirrhosis were collected. The nodular lesions were microdissected on hematoxylin and eosin-stained paraffin sections. Genomic DNA was isolated and pretreated with the methylation-sensitive enzyme Hpa Ⅱ or Hha Ⅰ, and amplified via nested PCR. The products were resolved on agarose gels and denaturing polyacrylamide gels. Hepatocellular carcinoma (HCC) and adenoma (HA) tissues were used as references. RESULTS: Monoclonality was demonstrated in all of the five cases of HCC and one case of HA. Totally 6 regular regenerative nodules and 29 NAHs including clear-cell (glycogen-storing) and mixed-cell types were microdissected from 4 cirrhotic liver specimens. Loss of X chromosomal inactivation mosaicism was demonstrated in all of the 5 mixed-cell NAHs with small-cell change (SCC), indicating their neoplastic nature. One of the lesions exhibited similar X chromosomal inactivation pattern as the neighboring HCC nodules, reflecting their common clonal origin. Among the 20 mixed-cell lesions without SCC, 7 (35%) were shown to be monoclonal, while all of the 4 clear-cell lesions and 6 regular regenerative nodules examined were found to be polyclonal. CONCLUSION: Some NAH lesions in the HBV-associated cirrhosis, particularly those with SCC, are already neoplastic lesions. Occurrence of SCC is a late event during NAH progression, a premalignant morphologic phenotype.
出处
《世界华人消化杂志》
CAS
北大核心
2005年第8期945-952,共8页
World Chinese Journal of Digestology
基金
国家自然科学基金资助项目
No 30171052~~
关键词
慢性乙型肝炎
肝硬化组织
变异肝细胞结节
克隆性
Clonality
Nodules of altered hepatocytes
Hepatitis B
Hepatic cirrhosis
Hepatocellular carcinoma
Premalignant lesions
