H1和H3型受体介导组胺双向调控肝癌细胞增殖的作用机制研究

Histamine (HA), a crucial autacoid in human, has been gaining remarkably importance for current tumor researches in these years. Previous reports indicate that HA generates its positive and negative regulation through binding to its four receptors (H1, H2, H3, H4). Our preliminary study suggests HA could activate H4 receptor and thus inhibits proliferation of pulmonary cancer cells. However, the effect and potential mechanism of HA and its receptors in hepatocellular carcinoma (HCC) remains to be investigated. Another pilot studies reveals that H1 and H3 are expressed in both HCC patient samples and HCC cell lines. Following study indicated the activation of H1 or H3 receptors in such cells could promote or inhibit their proliferation, respectively. But its underlying mechanisms needs further research. Based on these novel findings, our goal in this project is to characterize the relation between H1 and H3 expression and the pathological features or clinical prognosis in HCC. Taking advantage of histological examine on patient tissue samples, cells and tumor-planted mice model, and by using methods such as treatment of agonist and antagonist of HA receptors, overexpression and knockdown of their mRNA expressions, Quantification of cellular calcium, and protein chip, we could systematically analyze the distinctive roles of H1 and H3 in regulation of HCC cells. And thus provide novel insights and putative therapeutic target for treatment of such patients.

组胺（HA）作为一种重要的自体活性物质，在肿瘤研究中日益受到重视。以往文献报道，HA可激活四种不同类型HA受体（H1、H2、H3、H4）发挥正向或负向调控作用。我们在肺癌中的研究发现，HA可通过激活H4型受体抑制肺癌细胞增殖，然而HA及其受体在肝癌进展中发挥何种作用及其机制尚不明确。我们前期预实验结果表明，HA受体呈现明显的肝癌特异性表达模式，H1和H3型受体异质性高表达；初步细胞学实验表明激活H1或H3型受体，可显著促进或抑制肝癌细胞增殖，但其具体作用机制亟待阐明。基于上述发现，本项目拟在明确组胺H1和H3型受体表达与肝癌临床病理特征及预后相关性的基础上，借助组织标本、细胞和荷瘤小鼠模型，利用特异性激动剂和拮抗剂、过表达及RNA干涉、Ca2+荧光定量、蛋白抗体芯片等技术，系统分析H1和H3型受体在HA调节肝癌细胞增殖中的双向作用及分子机制，为肝癌防治提供新的理论支持和潜在分子靶点。

【研究背景】组胺发挥其多种生物学作用主要通过其四种同源G蛋白偶联组胺受体：H1HR，H2HR，H3HR和H4HR。最近研究发现，H1HR对肿瘤的增殖有一定影响。H3HR也参与了相关肿瘤的发生、增殖、转移等过程，胆管癌的体内、外实验表明组胺通过H3HR，激活IP3/Ca2+/PKCα通路，最终控制胆管癌细胞增殖。近期研究表明，组胺受体导致非酒精性脂肪肝的发展，表明组胺受体也可能在肝癌发生过程中发挥作用。然而，组胺受体在肝癌中潜在的功能作用还不清楚。在本研究中，我们对H1HR、H3HR在HCC病人组织样本中的表达情况进行了检测，然后对H1HR影响HCC细胞的生物学功能及分子机制进行了系统深入的研究。我们的研究有助于了解H1HR、H3HR在促进HCC进展中的作用机制，并为H1HR、H3HR作为肝癌潜在的治疗靶点提供了科学依据。.【研究内容】1、分析H1HR、H3HR在肝癌组织样本中的表达以及分析其与肝癌病人预后相关性。2、检测 H1HR 、H3HR对肝癌细胞生长的影响。3、检测H1HR、H3HR对肝癌细胞转移作用的影响。4、探究H1HR影响肝癌细胞生长的作用机制。5、鬼笔环肽染色检测H1HR、H3HR对肝癌细胞板状伪足的形成的影响，从而确定H1HR影响肝癌细胞转移的作用机制。6、分析H1HR影响肝癌细胞增殖和转移的作用机制。7、H1HR特异性拮抗剂Terfenadine检测其对肝癌细胞增殖和转移的作用影响，从而筛选出潜在治疗肝癌的靶向药物。.【研究结果】第一部分：研究结果显示肝癌病人中H1HR、H3HR是高表达且与肝癌的发展及肝癌病人的不良预后存在着重要的相关性。第二部分：H1HR、H3HR能够显著促进肝癌细胞的生长及转移。第三部分：H1HR能够加快细胞周期进程并抑制肝癌细胞凋亡从而促进肝癌细胞增殖且能够促进肝癌细胞板状伪足形成及MMP2的分泌从而促进肝癌的迁移和侵袭，并且最后研究得出：H1HR主要通过激活PKA信号通路进而促进了肝癌细胞的生长和转移。第四部分： H1HR抑制剂能够显著抑制肝癌细胞增殖和转移。.我们的研究结果表明，H1HR在肝癌细胞的生长和转移中起着关键作用，这为H1HR作为肝癌治疗的潜在药物靶点提供了新的科学理论依据。