从门静脉癌栓建立肝癌细胞株及对其细胞遗传特征的研究

Establishment and Cytogenetic Characterization of A Human Hepatocellular Carcinoma Cell Strain from the Embolus in Portal Vein

背景和目的:肝细胞癌(hepatocellularcarcinoma,HCC)预后差的原因是术后复发率高和早期通过门静脉转移。本研究建立门静脉癌栓肝癌细胞株,并探讨其分子细胞遗传特征。方法:从HCC患者门脉癌栓中获取肝癌细胞进行细胞培养,对培养成功的细胞(H4M)进行染色体G带染色后分析其核型和对比基因组杂交(comparativegenomichybridization,CGH)结果;用差异PCR检测周期素D1基因。结果:H4M细胞核型为超3倍体,染色体数为71～78,其中有一条标记染色体含有一长的均染区(hsr)。H4M主要的细胞遗传学改变为8p缺失和11q13高拷贝数扩增。周期素D1基因CCND1明显扩增。结论:染色体8p丢失和11q13高拷贝数扩增与H4M细胞的转移特性相关。周期素D1基因CCND1扩增可能是染色体11q13扩增的原因。

Background & Objective: Poor prognosis of hepatocellular carcinoma (HCC) is due to its high recurrent rate after operation and early metastasis through portal vein. This study was designed to establish a metastatic HCC cell line and to investigate its molecular and cytogenetic characterization. Methods: Cell culture was performed using the tissue obtained from a metastatic lesion in portal vein of a patient with HCC. After G banding staining,karyotype, and comparative genomic hybridization (CGH) of the cultured H4M cells were characterized and cyclin D1 gene CCND1 was detected by differential PCR. Results: The karyotype of metastatic HCC cell strain is a hypertriploid (71～78 chromosomes) with a huge marker chromosome containing a long homogeneously staining region (hsr). The main genetic alterations in H4M cells were a high copy number amplification of 11q13 and loss of 8p. Cyclin D1 gene CCND1 was also amplified significantly in the cells. Conclusions: The loss of chromosome 8p and high copy number amplification of 11q13 are associated with the metastatic characterics of H4M cells. The amplification of cyclin D1 gene CCND1 in H4M cells may be the cause of the amplification of chromosome 11q13.