异紫堇碱抑制携带人乳头瘤病毒16E6基因宫颈癌细胞增殖作用的机理研究

Cervical cancer is one of the most common cancers women have in our country. Human papilloma virus (HPV) infection will result in HPV geno combined to host cell and express relative HPV protein, which is the main mechnism leading to cervical cancer.However, currently there is no treatment for HPV infection in cervical cancer and the therapeutic effect is not that effective.It is likely that the intergration of traditonal Chinese medine with modern medical treatment will provide new methods for cervical cancer treatment. Our early studies have found that Chinese herbal monomer lsocorydine can inhibit HPV gene expression of SiHa、HeLa cell and facilitate apoptosis of cervical cancer cell,but the effects of lsocorydine played by inhibiting the activity of HPV cervical cancer cells are still not clear. Based on the preliminary studies, the project will conduct the following work by using animal experiments as well as cell experiment and by adopting moleculor biologic technologies and genomic, proteomic technologies: ①to validate the influence of Isocorydine HPV16E6 throuth inference experiment on SiHa cell of cervical cancer and cancer-bearing nude mice experiment; ②to analyze the molecular networks of the effects of Icorydaline in inhibiting cervical cancer by adopting transcriptomic, proteomic and other ways ; ③ to clarify the influence of Isocorydine on inhibiting cervical cancer cell cycle,cell apoptosis and HPV16E6 relative signaling pathway. The great significance of the present project is to provide a solid experimental and theoretical foundation for the possible effects of Isocorydine as a new and effective treatment of HPV infection and cervical cancer Chinese medicine unit and for designing a class of new drugs in this field.

宫颈癌是我国妇女最常见的恶性肿瘤,人乳头瘤病毒（HPV）感染是宫颈癌发生最主要的病因。目前尚缺乏针对抑制HPV而达到治疗宫颈癌的方法。如能利用传统中药与现代医学有机结合，将可能为宫颈癌防治提供新途径。我们前期工作发现中药单体异紫堇碱能够抑制宫颈癌SiHa、Hela细胞株HPV基因表达,并能有效促进癌细胞凋亡，但其相关机理仍不明确。本项目在前期研究基础上，拟通过细胞实验和动物实验，采用分子生物学、基因组学、蛋白质组学等技术，进行以下研究：①通过对宫颈癌Siha细胞株的干预和裸鼠荷瘤实验验证异紫堇碱对于HPV16E6的影响；②采用转录组和蛋白质组等组学手段分析异紫堇碱防治宫颈癌作用的分子网络；③阐明异紫堇碱对宫颈癌细胞周期抑制、凋亡等作用以及对HPV16E6相关信号通路的影响。本研究可以为异紫堇碱成为一种治疗HPV感染和宫颈癌的新的有效中药单体，或以其为基础设计一类新药提供实验依据和理论支持。

宫颈癌是我国妇女最常见的恶性肿瘤,人乳头瘤病毒（HPV）感染是宫颈癌发生最主要的病因。目前尚缺乏针对抑制HPV而达到治疗宫颈癌的方法。如能利用传统中药与现代医学有机结合，将可能为宫颈癌防治提供新途径。我们前期工作发现中药单体异紫堇碱能够抑制宫颈癌SiHa、Hela细胞株HPV基因表达,并能有效促进癌细胞凋亡，但其相关机理仍不明确。本项目在前期研究基础上，采用基因芯片、Western blotting 及免疫组化等技术手段进行细胞实验和动物实验研究以进一步阐明异紫堇碱抑制宫颈癌发生发展的机理。研究发现，经过异紫堇碱处理后的Siha细胞株以及裸鼠肿瘤实体组织中HPV16E6的表达水平明显下降，而P53，Caspase14, Clca2以及P21在基因以及蛋白质水平上均表达上调，PARP-Cleaved蛋白表达下调，与流式细胞学手段检测凋亡未见明显改变的结果一致；细胞周期结果显示在G0/S期发生阻滞，同时，E-cadherin表达也明显上调，以上结果表明异紫堇碱是通过促进Siha细胞向上皮细胞转化的途径来抑制宫颈癌的发生发展。