Ochiai T, Ueda K, Urata Y, Yamano T, Konishi E, Ogino A, Kawai K, Itoi H, Sonoyama T, Yamagishi H, Oka T, Ashihara T
Second Dept. of Surgery, Kyoto Prefectural University of Medicine, Japan.
Gan To Kagaku Ryoho. 1996 Jun;23 Suppl 2:182-8.
We investigated the cell clonality of 12 cases of female solitary hepatocellular carcinoma (HCC) that were associated with hepatitis virus infection. The clonal origin of HCC could be assessed by the method based on restriction fragment length polymorphism (RFLP) of X-chromosome-linked androgen receptor gene (AR) and phosphoglycerate kinase (PGK) gene, taking advantage of random inactivation of one of two X-chromosomes by methylation in females. We extracted DNA samples from both fresh and paraffin-embedded specimens of the same lesion as a source of DNA sample for polymerase chain reaction (PCR). Consequently, it was possible to use methylation-sensitive restriction enzymes and PCR to study differential methylation patterns among alleles of these genes for both DNA samples. The RFLPs of AR gene and PGK gene were found in eight of 12 cases and five of 12 cases, respectively. There were two cases which had no RFLPs in either AR gene or PGK gene. All cases of HCC which had RFLP in either AR gene or PGK gene demonstrated monoclonal origin of the tumor regardless of their histologic patterns.
我们研究了12例与肝炎病毒感染相关的女性孤立性肝细胞癌(HCC)的细胞克隆性。利用女性两条X染色体之一通过甲基化随机失活的特性,基于X染色体连锁雄激素受体基因(AR)和磷酸甘油酸激酶(PGK)基因的限制性片段长度多态性(RFLP)方法,可以评估HCC的克隆起源。我们从同一病变的新鲜标本和石蜡包埋标本中提取DNA样本,作为聚合酶链反应(PCR)的DNA样本来源。因此,对于这两种DNA样本,可以使用甲基化敏感的限制性内切酶和PCR来研究这些基因等位基因之间的差异甲基化模式。在12例病例中,分别有8例和5例发现了AR基因和PGK基因的RFLP。有2例在AR基因和PGK基因中均未发现RFLP。在AR基因或PGK基因中具有RFLP的所有HCC病例,无论其组织学模式如何,均显示肿瘤的单克隆起源。