HIF-2α通过调控Radil参与肝癌侵袭转移的基础研究

Hypoxia inducible factor 2 alpha (HIF-2α) may play an important role in cancer metastais, but its role in hepatocellular carcinoma (HCC) is not clear. We found that HIF-2α primes HCC metastasis into lung, and there were a lot of target genes involved in cell migration. Ras association and DIL domains (Radil) is shown to play a role in cell adhesion and migration, and maybe was HIF-2αtargte gene. We will construct plasmids to get clones with different HIF-2αlevel and explore its role in Huh7, MHCC97L, MHCC97H cell lines by wounding assay, transwell , adhesion assay; Changing Radil expressin levels by shRNA and expressing plasmids, then explore Rap1 signal pathway genes' levels such as integrin, FAK, ERK by real time PCR, western blot; we will find Radil distribution trends with HIF-2αand test its effect on migration ability, invasion ability by electric cell substrate impedance sensing (ECIS).We will monitor tumor growth, metastais by pear image, evaluate circulating number cells, metastasis diameter. Finally, we will stain Radil and evaluate its expression score in HCC tissue microarray,analyze the correlation of Radil to clinical pathological characteristics.

缺氧诱导因子-2 alpha（Hypoxia inducible factor 2 alpha,HIF-2α）在肿瘤转移、复发中发挥着重要作用。与HIF-1α相比，HIF-2α在肝癌转移、复发的研究较少，分子机制缺乏深入的阐明。本项目初步发现HIF-2α可以调节肝癌细胞运动，促进肝癌肺转移；有大量的靶基因调节细胞迁移，Radil是其中之一；Radil的表达水平与HIF-2α相关，是细胞运动、粘附必不可少的因子，推测HIF-2α可能通过Radil影响肝癌细胞的运动，进而影响肝癌的转移。本项目将在前期工作的基础上，利用高、低表达HIF-2α的肝癌细胞系，通过体内、外实验，探讨HIF-2α通过Radil影响肝癌侵袭、转移的分子机制，并通过大样本肝癌组织标本验证其临床意义，最终揭示HIFs在肝癌转移、复发中的作用，为深入认识肝癌转移、复发的分子机制提供理论依据。

多项证据表明缺氧诱导因子-2 alpha（hypoxia inducible factor 2 alpha,HIF-2α）可能在肿瘤转移、复发中发挥着非常重要的作用。与 HIF-1α相比，HIF-2 α在肝癌转移、复发的研究较少，分子机制缺乏深入的阐明。本项目发现 HIF-2α可以 促进肝癌肺转移，其靶基因可以调节细胞运动能力；Rap1的下游效应物Radil的表达水平与HIF-2 α相关，是细胞运动、粘附必不可少的因子，HIF-2α通过 Radil 影响肝癌细胞的运动而影响肝癌的转移。本项目利用高、低表达 HIF-2α的肝 癌细胞系，通过体内、外实验探讨了 HIF-2α通过 Radil 影响肝癌侵袭、转移的分子机制，并通过大样本肝癌组织标本验证其临床意义，最终探讨 HIFs 在肝癌转移、复发中的作用。