前期研究发现HPV16中E6和E7对LKB1具有抑制作用，并可介导HIF-1α上调肺癌细胞中葡萄糖转运蛋白GLUT1的表达，同时Sp1 mRNA及GLUT1 mRNA在肺癌支气管刷检细胞中过表达，然而E6和E7抑制LKB1后如何激活HIF-1α，以及HIF-1α又如何上调GLUT1的表达目前尚不清楚。我们推测E6和E7抑制LKB1后，解除了HIF-2α的磷酸化，使其通过去磷酸化恢复成HIF-1α，激活的HIF-1α同c-Myc竞争与转录因子Sp1结合，形成HIF-1α- Sp1复合体，活化的Sp1与GLUT1的启动子区结合，上调GLUT1的蛋白和mRNA表达。本项目拟在明确E6和E7抑制LKB1和HIF-1α上调GLUT1表达的基础上，构建与转染E6、E7、LKB1、HIF-1α及Sp1的质粒和SiRNA序列，双向调控上述基因，应用CoIP和ChIP方法分别检测HIF-1α与Sp1以及Sp1与GLUT1启动子区的结合状态，阐明E6和E7介导HIF-1α上调GLUT1表达以及促进肺癌细胞增殖、迁移及转移的分子机制，为HPV相关性肺癌的发生发展提供理论依据，为肺癌的预防和治疗提供潜在靶点。

According to previous work, HPV16 E6 and E7 obviously had inhibitory effect on LKB1, upregulated the expression of GLUT1 by mediating HIF-1α in lung cancer cells, and the overexpression of Sp1 mRNA and GLUT1 mRNA were in the bronchial brushing cells of patients with lung cancer. However, it is not clear that how to activate HIF-1α and upregulate the expression of GLUT1 by HIF-1α after E6 and E7 inhibited LKB1. We hypothesized that the phosphorylation of HIF-2α was released and recovered to HIF-1α after E6 and E7 inhibited LKB1, activated HIF-1α compete with c-Myc and bind with Sp1 forming HIF-1α-Sp1 complex, activated Sp1 bind with the promoter region of GLUT1, upregulated the expression of both protein and mRNA of GLUT1. The project is based on having made clear the role that both E6 and E7 inhibited LKB1 and HIF-1α upregulated the expression of GLUT1, intends to constructs and to transfect the plasmid and the SiRNA sequence of E6, E7, LKB1, HIF-1α and Sp1, regulates the above genes by bidirection, CoIP and ChIP methods are used to detect the binding states of HIF-1α and Sp1as well as Sp1 and GLUT1 promoter regions respectively, elucidates the molecular mechanism of both E6 and E7 upregulated the expression of GLUT1 by mediating HIF-1α and of promoting the proliferation, migration and metastasis of lung cancer cells, provides theoretical basis for the occurrence and development of HPV related lung cancer and potential targets for prevention and treatment of lung cancer.