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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.
OBJECTIVE: To investigate the clonality of mutated hepatocyte nodules (NAH) in cirrhotic cirrhotic tissues using cloned phosphoglycerate kinase and androgen receptor site-based clonality assay based on X-chromosome inactivation chimerism in somatic cell tissues of female. Methods: Hepatitis B virus (HBV) -related liver cirrhosis specimens were paraffin-embedded, HE staining was used to obtain hepatocyte nodules by microdissection. Genomic DNA was extracted and digested with methylation-sensitive HpaII or HhaI. , Gel electrophoresis showed the results.Application of hepatocellular carcinoma (HCC) and hepatocellular adenoma as a reference lesion.Results: HCC5 cases and hepatocellular adenoma tissue were monoclonal.Among four cases of analysis of cirrhosis Tissues were isolated from the common regenerative nodules 6 and NAH 29, the latter including mixed cell lesions 25, transparent cell (glycogen storage) lesions 4. 5 small cells associated with small cell changes (SCC) mixed cells Sexual NAH showed X chromosome inactivation chimerism lost, suggesting that the tumor lesions; one of the lesion taken adjacent to the tumor and inactivation of the same type of band, and HCC have the same proven origin of cloning; 20 do not Of mixed-cell NAH with SCC, 7 (35%) were single Long lesions; The detection of 4 clear cell NAH and 6 normal regenerative nodules did not show loss of X chromosome inactivation chimerism, suggesting polyclonal lesions.Conclusion: HBV-related part of cirrhexis NAH, especially lesions associated with SCC, is monoclonal and is already a neoplastic lesion; SCC is a more recent change in the progression of NAH and is a precancerous lesion.