长链非编码RNA（LncRNA-BX648637)调控细胞粘附因子1(ICAM-1）促进肝癌门静脉癌栓形成的机制研究

The portal vein tumor thrombus (PVTT) is one of the main factors which induce the poor prognosis of hepatocellular carcinoma，however, the mechanism of PVTT is still unknown now.In order to verify the different expression of lncRNAs and mRNAs between PVTT tissues and parenchyma tumor (PT) tissues, we first performed a gene expression profiling array examination between CSQT-2 and Hep3B cell lines. Many lncRNAs and mRNAs were observed to be high expressed in CSQT-2 cell line.Among these RNAs, we selected some co-expression lncRNAs and mRNAs for further research. We found that there were 812 complementary base pairs between the sequences of LncRNA-BX648637 and ICAM1, so we supposed that LncRNA-BX648637 may regulate ICAM1 to contribute the formation of PVTT. In order to conform our hypothesis, we will use qRT-PCR to validate the different expression of ICAM1 and BX648637 between PVTT and PT tissues. Next, we will identify the relationship of the expression of ICAM1 and BX648637 through an RNase protection assay (RPA).Finally about 300 patients with different typle of PVTT will be tested the relationship between lncRNA-BX64837 and prognosis of HCC. This study may reveal that lncRNA-BX648637 regulate ICAM-1 to contribute the formation of PVTT, and lncRNA-BX648637 maybe a new molecular marker and a potential therapeutic target for the HCC patients with PVTT.

门静脉癌栓（PVTT）是影响原发性肝癌预后最重要的因素之一，至今其发生机制仍不清楚。研究PVTT发生机制并进一步寻找干预手段对于提高肝癌整体疗效至关重要。本课题前期利用LncRNA基因芯片和mRNA芯片技术，发现LncRNA-BX648637和ICAM1 在癌栓细胞系（CSQT-2）和癌栓组织上都高表达，而且两者之间存在812对互补碱基序列。因此我们推测LncRNA-BX648637可能调控ICAM1促进PVTT形成。本研究拟通过分子生物学实验证实lncRNA能够调控ICAM1促进门静脉癌栓形成，并通过动物模型加以验证，最后通过大量临床资料验证LncRNA与不同类型PVTT患者预后存在密切联系。本研究有望将LncRNA-BX648637作为PVTT潜在诊断和干预靶标，从而为PVTT的诊断和治疗提供新的途径。

研究背景和目的. 肝癌合并门静脉癌栓（PVTT）是影响肝癌预后的主要因素之一，PVTT的形成是一系列多环节、多因素复杂过程，目前其发生机制仍未完全清楚。长非编码RNA(lncRNAs)参与肝癌的发生发展,但是lncRNAs和肝细胞癌门静脉癌栓转移之间的关系尚不清楚。如今,越来越多的研究表明,肿瘤干细胞在肿瘤侵袭转移中也起到关键作用。我们之前的研究发现,ICAM1 +细胞具有肿瘤干细胞能力，并参与肝癌的发生和转移显著相关。我们前期利用基因芯片技术发现lncRNA－648637和ICAM-1在肝癌门静脉癌栓细胞系和癌栓组织中都存在高表达,且两者存在812互补碱基序列，因此我们猜测LncRNA-64863是否通过调控细胞粘附因子1（ICAM-1）阳性肿瘤干细胞促进门静脉癌栓转移。.研究内容. 我们测量了LncRNA - 648637和ICAM1在原发性肝癌原发肿瘤组织和相应的门静脉血栓(PVTT)组织中的表达水平。我们也比较了LncRNA - 648637和ICAM1 CSQT-2细胞(PVTT细胞系)和Hep3B细胞（肝癌细胞系）上的表达水平。我们 利用ROC生存曲线进行了分析肝癌合并门静脉癌栓的预后相关因素,探索肿瘤中LncRNA - 648637 / ICAM1表达与肝癌相关的预后意义。我们评估了肿瘤发生的潜在的ICAM1 +体外肝癌细胞系,并利用动物模型证实LncRNA-648637是否通过调控细胞粘附因子1（ICAM-1）阳性肿瘤干细胞促进门静脉癌栓转移。.重要结果. LncRNA-648637与ICAM1能够形成稳定双联,这增加了ICAM1＋肿瘤干细胞的特性。LncRNA-648637和ICAM1表达水平晚期肝癌PVTT更是高表达。肿瘤组织上高表达LncRNA-648637与肝癌的转移和复发密切有关。此外, LncRNA-648637与ICAM1能够形成稳定双联促进ICAM1 + 肝癌肿瘤干细胞的特性促进肝癌门静脉癌栓形成。.科学意义. LncRNA-648637通过调控ICAM1＋细胞，促进肝癌肿瘤干细胞特性，促进肝癌门静脉癌栓的形成，同时也是肝癌合并门静脉癌栓复发和生存的独立预后因素。因此, LncRNA-648637可以肝癌合并PVTT新的诊断标记物和干预靶标。