融合基因HPV-KLF12在HPV整合导致宫颈癌发病中的作用及机制研究

高危型人乳头瘤病毒(high risk Human Papillomavirus；hrHPV)持续性感染是宫颈癌的主要致病因素， hrHPV人类基因组整合引起的基因组结构不稳定是宫颈癌发生最为关键的分子事件之一。我们前期围绕“hrHPV持续性感染与整合”利用三维测序等技术开展了系统深入的研究，并利用CRISPR/Cas9技术在正常鳞状上皮Hacat细胞染色体13q22.1位点插入HPV16 E6和E7序列，构建HPV整合细胞模型。本项目拟在体内外验证细胞模型并观察细胞恶性转化的生物学表型，采用三维测序技术（Hi-C）,RNA-seq联合分析HPV整合引起的人类基因组结构变化以及RNA水平产生的融合基因。在此基础上，深入研究融合基因HPV-KLF12在宫颈癌恶性转化中的作用及机制。该项目是既往HPV整合产生融合基因的研究以及三维测序研究的深入和创新，具有一定的理论意义和临床应用前景。

High risk human papillomavirus (hrHPV) persistent infection is the main pathogenic factor of cervical cancer. In the process of hrHPV persistent infection, the instability of genome structure caused by HPV integration of human genome is one of the most critical events of cervical carcinogenesis. We carried out systematic and in-depth research on "hrHPV integration and persistent infection” by using three-dimensional sequencing in previous studies, and inserted HPV16 E6 and E7 sequences into the chromosome 13q22.1 of normal squamous epithelial HaCaT cells by using CRISPR / cas9 technology to build an integrated cell model of HPV. This project intends to verify the HPV integrated cell model in vitro and in vivo and observe the biological phenotype of malignant transformation of cells. Based on above observation, in the integrated cell model Hacat-HPV+, three-dimensional sequencing (Hi-C) and RNA-Seq will be applied to analyze the structural changes of human genome caused by HPV integration as well as the RNA level fusion genes produced by structural changes. Moreover, further role and mechanism of HPV-KLF12 fusion gene in cervical cancer malignant transformation will be carried out. This project is an in-depth and innovative research on the fusion gene generated by HPV integration and 3D sequencing, which has certain theoretical significance and important clinical application prospects.