论文部分内容阅读
循环肿瘤细胞(CTCs)因携带原发肿瘤的一些生物特性以及自身独特的生物学功能,在癌症的诊断、治疗和转移预防方面发挥着重要作用。但外周血中CTCs的低含量使其检测、分离和富集有一定技术难度。本文选取结肠癌HT29细胞为CTCs模型,以表面高表达的Ep CAM为生物标记物,选取纳米级高分子聚合物G6 PAMAM dendrimers为基质,多价络合针对Ep CAM的抗体来构建络合物,实现体外对HT29细胞的特异性捕获及活性调控。应用红外、动态光散射、紫外和荧光等方法表征络合物的理化特性,荧光分析法考察络合物对HT29细胞的识别和捕获,MTT、荧光拍照、流式分析法显示络合物对捕获的HT29细胞的活性调控。结果表明,制备的络合物可以高特异性地识别和结合贴壁的和悬浮的HT29细胞,从而对捕获的细胞产生一定的活性下调作用,但不产生严重的细胞毒效应。可见,利用具有多价络合效应的纳米材料来共价连接针对CTCs表面标记物的靶向抗体,有望实现在病人血中对CTCs的特异性捕获和活性调控。
Circulating tumor cells (CTCs) play an important role in the diagnosis, treatment and metastasis prevention of cancer because of some biological characteristics of carrying primary tumors and their own unique biological functions. However, low levels of CTCs in peripheral blood make it difficult to detect, separate and enrich CTCs. This study selected colon cancer HT29 cells as CTCs model, with high surface expression of Ep CAM as a biomarker, selected nano-polymer G6 PAMAM dendrimers as a matrix, multivalent complex against Ep CAM antibody to construct complexes, To achieve specific HT29 cell in vitro capture and activity control. The physical and chemical properties of the complexes were characterized by infrared, dynamic light scattering, ultraviolet and fluorescence spectroscopy. Fluorescence analysis was used to identify and capture the HT29 cells. MTT, fluorescence photographs and flow cytometry HT29 cell activity regulation. The results showed that the prepared complexes could recognize and bind HT29 cells with high specificity and affinity to trap the cells, but did not produce serious cytotoxicity. Thus, the use of multivalent complexing nanomaterials to covalently link targeted antibodies against CTCs surface markers is expected to achieve specific capture and activity control of CTCs in the patient’s blood.