Clonal diversity and its imphcution in multistage development of hepatocellnlar caronoma

通过分析肝细胞癌患者和1例活体供肝原位肝移植患者的多发性肝结节的染色体改变，探讨基因组多样性及其在多阶段发育中的意义。肿瘤和非肿瘤组织在手术时获得肿瘤和非肿瘤组织，采用常规的蛋白酶K/酚-氯仿法提取DNA，用400条几乎均匀分布在22个常染色体和X染色体上的扩增微卫星基因座的方法提取DNA。检测肿瘤标本的聚合酶链式反应产物曲线下方区域的等位基因不平衡，并用非肿瘤标本单核苷酸多态性(SNP)芯片区分染色体和丢失。2004年4月至2007年8月，72例肝细胞癌患者接受了活体供肝移植。其中8例患者共获得20个结节。发现-1p、+1q、+5、+7、-8p、+8q、-16q、-17p等重复性染色体改变。染色体关系分析显示，8例患者中有7例为转移性结节，仅1例为多中心起源的肿瘤。此外，对30个结节的染色体分析发现，既有遗传稳定的肿瘤，后有常见的染色体改变，如-1p，+1q，-6q，-16q，+22q，也有不稳定的肿瘤，几乎所有的染色体都有多处改变。尽管在终末期肝病患者中发现的肝细胞癌患者的染色体改变总体上是稳定的，但染色体不稳定的发生机制可能是未知的，但可能是不同的机制。

Genomic diversity and its implication in multistage development were explored by analyzing chrmosomal alterations found in multiple liver nodule obtained from patients with hepatocellular carcinoma who had and one living donor orthotopic liver transplantation.Tumor and non tumor tissue were obtained at the time of surgery, from patients who had provided written informed consent DNA was extracted by using conventional proteinase K/phenol-chloroform method Four-hundred pane of PCR primers that distribute almost equally throughout 22 autosome and X chromosome wee used to amplify microsatellite loci. Allelic imbalances were determined on the bass of the area under the curve of PCR product that derived from the tumor and the non-tumor sample Single nucleotide polymorphism (SNP) array were used to discriminate chromosomal and losses.Seventy-two patients with hepatocellular carcinoma had received living-donor liver transplantation in between April 2004 and August and August 2007. A total of 20 nodules were obtained from eight of these patients. Recurrent chromosome alterations, such as -1p, +1q, +5, +7, -8p, +8q, -16q, -17p, were detected. Chromosomal relationship analysis revealed that seven patients out of eight had metastatic nodule, while only one patient had tumors of multicentric origin.Furthermore, chromosome analysis of 30 nodules m a patient detected both genetically stable tumors that was hind to have common chromosome alterations such as -1p, +1q, -6q, -16q, +22q, and unstable tumors that had multiple alterations throughout almost all chromosome.Although chromosomal alterations detected m hepatocellular carcinoma m patients with end-stage liver disease were generally stable, chromosomal instability might occur by an unknown but possibly different mechanism.