人疱疹病毒6型U94/rep基因的表达在调控人神经胶质瘤发展中的作用

In 2009,we isolated a kind of virus from cycst fluid of glioma, which was identified as HHV-6A by virology and molecular biology，It was the first time to report this finding. HHV-6 U94/rep,which is homologous with AAV-2 rep gene，HHV-6 U94/rep gene can inhibit the formation of lymphatic vessels and vascular endothelial cells in vivo and in vitro.The preliminary experiments found that the glioma cells expressing U94/rep gene, whose proliferation, cloning, cell cycle progression and vascular generation capabilities were been suppressed.This study contains (1)The effects of U94/rep gene for the biological function of glioma cells in vivo and in vitro will be observed by using gene transfection and RNA interference technology. (2) To explore the production of a variety of chemokine and chemokine receptor expression of HHV-6 infected glia cells and the anti-glioma effect of selected the key chemokines.（3）The results of clinical specimens testing will reveal the correlation between the expression of U94/rep and clinical glioma progression and prognosis.These results will not only clarify the molecular mechanism of U94/rep regulating glioma and HHV-6 inducing chemokines and chemokine receptors expression, but also provide the basis for treatment and preventive anti-glioma vaccines.

2009年课题组从胶质瘤囊液中鉴定到1株HHV-6A，属国内外首次。HHV-6的转化抑制基因U94/rep与AAV-2 rep具有同源性，可以抑制体内外淋巴管和血管内皮细胞的形成。预实验发现U94/rep可以抑制胶质瘤细胞增殖、克隆、细胞周期进程及促血管生成能力。本研究拟（1）运用基因转染、RNA干扰等技术在体内及体外实验中观察U94/rep基因对胶质瘤细胞生物功能的影响。（2）探讨HHV-6感染刺激神经胶质细胞产生多种趋化因子及其受体，调控关键性趋化因子在抗胶质瘤中的作用。（3）临床标本检测，揭示U94/rep基因表达水平与胶质瘤进展及预后的相关性。本研究结果将阐明U94/rep调控胶质瘤细胞及HHV-6诱导趋化因子及受体表达在抗胶质瘤发病中的分子机制，为研究治疗及预防性抗胶质瘤疫苗提供依据。

已知U94/rep在HHV6潜伏感染中是高表达的基因，本课题主要研究HHV6 U94/rep基因在抑制神经胶质瘤中的作用及主要机制：1.首次建立了三个稳定表达U94/rep基因的U251细胞株；U94/rep基因的U87细胞株；人脐静脉血管内皮细胞EAhy926-U94/rep细胞株。运用此三个细胞株对细胞的增殖、细胞克隆形成、划痕试验、transwell迁移试验、血管形成试验、细胞周期等生物学功能的影响，证实了U94/rep基因有抑制胶质瘤细胞的功能；2.建立了裸鼠的动物模型，直接观察到肿瘤形成的大小、重量都区别于对照组；3.为了寻找U94/rep基因能抑制神经胶质瘤细胞的关键部分，将U94/rep切割成二部分，初步结果显示U94抑瘤功能主要存在于前部分，此研究还需进一步探讨；4.我们探讨了胶质瘤及其癌旁组织基因表达谱的变化；5.染色体整合的HHV6调研，因ci-HHV6对脑、心血管疾病有重要的诊断和治疗应用前景，我们在国内首次进行，已研制成试剂盒，并已拿到1项专利，已获1项国际资金资助。