Establishment and cytogenetic characterization of a human hepatocellular carcinoma cell strain from the embolus in portal vein

Abstract

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 characteristics of H4M cells. The amplification of cyclin D1 gene CCND1 in H4M cells may be the cause of the amplification of chromosome 11q13.

背景和目的:肝細胞癌(hepatocellularcarcinoma,HCC)預后差的原因是術后復發率高和早期通過門靜脈轉移。本研究建立門靜脈癌栓肝癌細胞株,并探討其分子細胞遺傳特征。方法:從HCC患者門脈癌栓中獲取肝癌細胞進行細胞培養,對培養成功的細胞(H4M)進行染色體G帶染色后分析其核型和對比基因組雜交(comparativegenomichybridization,CGH)結果;用差異PCR檢測周期素D1基因。結果:H4M細胞核型為超3倍體,染色體數為71～78,其中有一條標記染色體含有一長的均染區(hsr)。H4M主要的細胞遺傳學改變為8p缺失和11q13高拷貝數擴增。周期素D1基因CCND1明顯擴增。結論:染色體8p丟失和11q13高拷貝數擴增與H4M細胞的轉移特性相關。周期素D1基因CCND1擴增可能是染色體11q13擴增的原因。