病毒表达和肝组织对乙型肝炎病毒显微肝细胞的分子表达谱 -

StephenW

Viral expression and molecular profiling in liver tissue versus microdissected hepatocytes in hepatitis B virus - associated hepatocellular carcinoma

Marta Melis1, Giacomo Diaz2, David E Kleiner3, Fausto Zamboni4, Juraj Kabat5, Jinping Lai3, Giulia Mogavero4, Ashley Tice1, Ronald E Engle1, Steven Becker5, Charles R Brown6, Jeffrey C Hanson7, Jaime Rodriguez-Canales7, Michael Emmert-Buck7, Sugantha Govindarajan8, Michael Kew9 and Patrizia Farci1*   

    * Corresponding author: Patrizia Farci [email protected]

Author Affiliations



Journal of Translational Medicine 2014, 12:230  doi:10.1186/s12967-014-0230-1
Published: 21 August 2014
Abstract
Background

The molecular mechanisms whereby hepatitis B virus (HBV) induces hepatocellular carcinoma (HCC) remain elusive. We used genomic and molecular techniques to investigate host-virus interactions by studying multiple areas of the same liver from patients with HCC.
Methods

We compared the gene signature of whole liver tissue (WLT) versus laser capture-microdissected (LCM) hepatocytes along with the intrahepatic expression of HBV. Gene expression profiling was performed on up to 17 WLT specimens obtained at various distances from the tumor center from individual livers of 11 patients with HCC and on selected LCM samples. HBV markers in liver and serum were determined by real-time polymerase chain reaction (PCR) and confocal immunofluorescence.
Results

Analysis of 5 areas of the liver showed a sharp change in gene expression between the immediate perilesional area and tumor periphery that correlated with a significant decrease in the intrahepatic expression of HB surface antigen (HBsAg). The tumor was characterized by a large preponderance of down-regulated genes, mostly involved in the metabolism of lipids and fatty acids, glucose, amino acids and drugs, with down-regulation of pathways involved in the activation of PXR/RXR and PPARα/RXRα nuclear receptors, comprising PGC-1α and FOXO1, two key regulators critically involved not only in the metabolic functions of the liver but also in the life cycle of HBV, acting as essential transcription factors for viral gene expression. These findings were confirmed by gene expression of microdissected hepatocytes. Moreover, LCM of malignant hepatocytes also revealed up-regulation of unique genes associated with cancer and signaling pathways, including two novel HCC-associated cancer testis antigen genes, NUF2 and TTK.
Conclusions

Integrated gene expression profiling of whole liver tissue with that of microdissected hepatocytes demonstrated that HBV-associated HCC is characterized by a metabolism switch-off and by a significant reduction in HBsAg. LCM proved to be a critical tool to validate gene signatures associated with HCC and to identify genes that may play a role in hepatocarcinogenesis, opening new perspectives for the discovery of novel diagnostic markers and therapeutic targets.
Keywords:
Hepatocellular carcinoma; Hepatitis B virus; Gene expression profiling; Laser capture microdissection; Confocal microscopy

StephenW

病毒表达和肝组织对乙型肝炎病毒显微肝细胞的分子表达谱 - 相关肝癌

玛塔Melis1，贾科莫Diaz2，大卫êKleiner3，福斯托Zamboni4，尤拉伊Kabat5，锦屏Lai3，朱Mogavero4，阿什利Tice1，罗纳德·êEngle1，史蒂芬Becker5，查尔斯ŕBrown6，杰弗里çHanson7，海梅·罗德里格斯Canales7，迈克尔·埃默特-Buck7， Sugantha Govindarajan8，迈克尔Kew9和帕特里齐亚Farci1*

    *通讯作者：帕特里齐亚Farci [email protected]

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杂志转化医学2014年，12：230 DOI：10.1186/ s12967-014-0230-1
发布时间：2014年8月21日
摘要
背景

的分子机制，使B型肝炎病毒（HBV）引起肝细胞癌（HCC）仍然是难以捉摸的。我们用基因组学和分子技术从患者的肝癌研究同一肝脏的多个领域探讨宿主 - 病毒相互作用。
方法

我们比较了全肝组织（WLT）的基因标记与激光捕获显微切割（LCM）的肝细胞随着乙肝病毒的肝内表达。基因表达谱进行多达在从11例HCC患者个体的肝脏肿瘤中心不同距离获得17 WLT标本和所选LCM样本。在肝脏和血清HBV标记物通过实时聚合酶链反应（PCR）和共聚焦免疫荧光测定。
结果

肝脏的5个区域的分析表明，直接损伤周围的区域和肿瘤周边与在HB表面抗原（HBsAg）的肝内表达显著减少相关之间的急剧变化中的基因表达。肿瘤的特点是下调的基因，主要参与脂类和脂肪酸，葡萄糖，氨基酸和药物的代谢大优势，具有下调参与PXR/ RXR和PPARα/RXRα的激活途径的核受体，其包括，PGC-1α和FOXO1，两个关键调节危重不仅参与肝脏的代谢功能，而且在HBV的生命周期中，作为必需的转录因子的病毒基因表达。这些结果由显微肝细胞的基因表达的证实。此外，恶性肝细胞的LCM还透露上调与癌症和信号传导途径，包括两种新型肝癌相关的癌睾丸抗原基因，NUF2和TTK相关的独特基因。
结论

全肝组织的显微肝细胞的整合基因表达分析表明，HBV相关HCC的特征是代谢关断和通过显著减少的HBsAg。 LCM证明，以验证与肝癌相关基因标记，并确定基因，可能在肝癌中发挥作用的重要工具，对于新的诊断标志物和治疗靶点的发现开辟了新的视角。
关键词：
肝癌;乙型肝炎病毒;基因表达谱;激光捕获显微切割;共聚焦显微镜