构建基于TRAIL-exosome的siRNA和Cisplatin共载纳米系统及治疗耐药宫颈癌的研究

Cervical cancer is one of the most common gynecologic malignancies. Cisplatin resistance is a difficult problem to overcome in the chemotherapy of cervical cancer. Researches showed that exosome was considered to be one of the most ideal chemotherapy drug nanocarriers attributed to its low immunogenicity and good homing ability. Therefore, this project intends to develop an exosome with its membrane expressing TRAIL protein on the basis of plasmid transfection by lentiviral systems which we have constructed, and then to load Bcl-2-siRNA and Cisplatin into the exosome, finally forming a TRAIL-exosome@Bcl-2-siRNA&Cisplatin co-loaded nanosystem. We will verify through vitro experiment that the nanosystem can effectively improve the sensitivity of drug-resistant cervical cancer cell line (SiHa/CDDP) to Cisplatin, and enhance Cisplatin inhibitory effect on tumor. Furthermore, we will also verify this nanosystem's biological compatibility in immune normal mice, and its in vivo tumor targeting and tumor inhibitory effect in the cervical SiHa/CDDP carcinoma subcutaneous transplantation tumor and in situ tumor animal model which we have already successfully constructed, and provide new ideas for the effective treatment of cisplatin resistant cervical cancer.

宫颈癌是最常见的妇科恶性肿瘤之一，克服顺铂（Cisplatin）耐药是宫颈癌化疗中的难题。研究表明，外泌体（exosome）由于较低的免疫原性及良好的归巢能力被认为是最理想的化疗药物纳米载体之一。因此，本项目拟在本课题组前期构建的质粒转染（慢病毒体系）的基础上开发一种膜上表达TRAIL蛋白的exosome，并在该exosome中装载Bcl-2-siRNA和Cisplatin，形成TRAIL-exosome@Bcl-2-siRNA&Cisplatin共载纳米系统。通过体外实验验证该纳米系统能有效提高Cisplatin对耐药的宫颈癌细胞SiHa/CDDP的肿瘤抑制效果。同时，验证该纳米系统在免疫正常小鼠中的生物相容性，并在本课题组已经成功构建的SiHa/CDDP宫颈癌皮下移植瘤和原位瘤的动物模型中验证该纳米系统对体内肿瘤的靶向性及肿瘤抑制效果，为有效治疗Cisplatin耐药宫颈癌提供新思路。

宫颈癌是最常见的妇科恶性肿瘤之一，克服顺铂（Cisplatin，DDP）耐药是宫颈癌化疗中的难题。以顺铂为基础的纳米系统能在一定程度上解决宫颈癌顺铂治疗中存在的靶向能力差、毒副作用强以及耐药等问题。本研究1.通过培养不同组织（脂肪、脐带、羊膜、绒毛膜）来源的间充质干细胞，观察各种间充质干细胞对宫颈癌细胞的趋化差异，发现绒毛膜来源的间充质干细胞对宫颈癌细胞的趋化作用最强，该作用和绒毛膜来源的间充质干细胞中含有高水平的CXCR4以及宫颈癌细胞分泌CXCL12密切相关，是由CXCL12/CXCR4通路介导。2.成功分离和鉴定间充质干细胞来源的膜上表达TRAIL蛋白的外泌体（exosome），并构建TRAIL-exosome@Bcl-2-siRNA&Cisplatin共载纳米系统，在体内、外实验中验证该纳米系统能有效提高耐药宫颈癌细胞对顺铂的敏感度，增强顺铂的肿瘤抑制效果。3.成功构建Cisplatin共载纳米载药体系（DDP-Au@MSN-Avatin@PEI/AE105），通过体内宫颈癌裸鼠皮下移植瘤和体外细胞实验均发现该复合纳米载药体系具有良好的稳定性、生物相容性和宫颈癌细胞靶向性，可通过联合光热治疗，抑制癌细胞的增殖，促进细胞周期阻滞以及线粒体断裂，同时，通过抑制瘤体微血管的生存、促进细胞凋亡等作用来有效抑制瘤体的生长。该研究为靶向及有效治疗宫颈癌及提高顺铂耐药宫颈癌的疗效提供帮助。