Expression of HNFs and C/EBPα is correlated with immunocytochemical differentiation of cell lines derived from human hepatocellular carcinomas, hepatoblastomas and immortalized hepatocytes
Expression of HNFs and C/EBPα is correlated with immunocytochemical differentiation of cell lines derived from human hepatocellular carcinomas, hepatoblastomas and immortalized hepatocytes
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DOI：
10.1111/j.1349-7006.2003.tb01515.x
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发表时间：
2003-09-01
期刊：
影响因子：
5.7
通讯作者：
Noguchi, M
中科院分区：
文献类型：
--
作者：
Ishiyama, T;Kano, J;Noguchi, M
肝细胞癌（HCC）分化程度的客观评价对HCC的病理诊断、预后及治疗方案的评价具有重要意义。肝细胞的分化通过其在小鼠胚胎中的肝功能蛋白的表达来反映，并且肝富集转录因子（liver-enriched transcription factors，LETF）已被证明严格地调节肝功能基因。以前的报道表明，LETF的表达水平在肝癌或癌前结节与非癌组织相比发生了改变。因此，LETF表达水平可能是有用的HCC成熟的措施。本研究采用实时荧光定量逆转录聚合酶链反应（real-time reverse transcription PCR，RT-PCR）和免疫细胞化学方法，对12株肝癌细胞系中HNF和C/EBPalpha mRNA的表达进行定量分析，以探讨LETF的表达与肝癌分化程度的关系。此外，我们研究了C.B-17/Icrj-scid/scid小鼠原位移植的肝癌细胞系，并描述了肿瘤的组织学和细胞学分化。我们的研究结果表明，HNF-3 β，HNF-4 α，HNF-1 α和C/EBPalpha的综合表达是特异性的肝癌分化良好的功能和形态。此外，在这四种转录因子中，HNF-4 α和HNF-1 α的表达具有同步性，并与HCC分化程度密切相关。这些体外结果在SCID小鼠体内形成的肿瘤中得到证实。这些结果表明HNF-4 α和HNF-1 α是评估HCC分化程度的有用标志物，我们建议通过定量分析HCC中HNF和C/EBPalpha来客观地评估HCC的分化程度。
Objective assessment of the differentiation grade of hepatocellular carcinomas (HCCs) is important for evaluation of the pathological diagnosis, prognosis and therapeutic treatment. Differentiation of hepatocytes is reflected by their expression of hepatic functional proteins in the mouse embryo, and liver-enriched transcription factors (LETFs) have been shown to regulate hepatic functional genes strictly. Previous reports demonstrated that the level of LETF expression is altered in HCC or preneoplastic nodules compared with noncancerous tissues. Therefore, LETF expression levels might be useful as a measure of HCC maturation. In this study, to clarify the correlation between the expression of LETFs and the differentiation grade of HCCs, we performed a quantitative analysis of the mRNA expressions of HNFs and C/EBPalpha using real-time reverse-transcription PCR and immunocytochemical analysis for hepatic functional proteins in twelve cell lines. Furthermore, we examined orthotopic transplantations of the HCC cell lines in C.B-17/Icrj-scid/scid mice and characterized the histologic and cytologic differentiation of the tumors that developed. Our results showed that comprehensive expressions of HNF-3beta, HNF-4alpha, HNF-1alpha, and C/EBPalpha were specific to HCCs with well-differentiated function and morphology. Furthermore, among these four transcription factors, HNF-4alpha and HNF-1alpha expressions showed synchronism and had a close relation with HCC differentiation. These in vitro results were confirmed in tumors developed in SCID mice in vivo. These findings suggested that HNF-4alpha and HNF-1alpha are useful markers to assess the degree of HCC differentiation, which we suggest could be evaluated objectively by the quantitative analysis of HNFs and C/EBPalpha in HCCs.