结肠癌HT29多细胞球的培养及其结构与生物学特性的观察  

Structure and biological features of multicellular spneroids of colon cancer HT29 cells

作　　者：何建明[1] 江恒[1] 陈克力[1] 黄海辉[1] 庞学利[1] 梁后杰[1] 

机构地区：[1]第三军医大学西南医院肿瘤科,重庆400038

出　　处：《第三军医大学学报》2010年第16期1691-1693,共3页Journal of Third Military Medical University

基　　金：国家自然科学基金(30171067);重庆市自然科学基金(CSTC2009BB5339);第三军医大学青年创新人才基金(2009XQN32)~~

摘　　要：目的建立结肠癌体外三维细胞培养模型HT29多细胞球,鉴定并检测其基本生物学特性。方法琼脂糖抑制细胞贴壁、水平回旋振荡法培养HT29多细胞球;扫描电镜、HE染色、免疫组化检测多细胞球的组织病理学性质;透射电镜检测细胞超微结构;Cell counting kit-8检测药物敏感性。结果成功培养由细胞相互黏附形成的多层细胞构成的结肠癌多细胞球模型;多细胞球内细胞接触广泛而紧密,可见大量细胞连接;多细胞球局部可见腺腔样结构,细胞角蛋白表达强弱与腺腔样结构有关;多细胞球药物敏感性显性降低,在500μg/L奥沙利铂处理48h条件下,单层细胞生存分数为(9.3±2.0)%,而多细胞球则为(42.9±4.0)%,二者相比差异有统计学意义(P<0.05)。结论成功建立了结肠癌多细胞球模型,此模型是研究体内无血管的微小结肠癌及结肠癌无血管区的较理想模型。Objective To establish a three-dimensional colon cancer cells culture model,multicellular spheroids for HT29 cells and detect their basic biology.Methods Multicellular spheroids of HT29 cells were cultured in flasks coated with 2% agarose by gently convoluting horizontally.Scanning electron microscopy,HE staining and immunohistochemistry were used to detect the histopathology of HT29 multicellular spheroids,and transmission electron microscopy was used to detect the ultrastructure.Drug sensitivities of both HT29 monolayer and HT29 multicellular spheroids were evaluated by using cell counting kit-8.Results HT29 multicellular spheroids consisted of layers of cells which adhered with each other closely and cell-cell junctions were found in most cells in multicellular spheroids.Lumen-like of glands were found in multicellular spheroids.Expression level of cytokeratin was related with lumen.HT29 cells acquired multicellular drug resistance when grown as multicellular spheroids.Cell survival fraction of HT29 monolayers was （9.3±2.0）% when they were treated with 500 μg/L oxaliplatin for 48 h,while that of multicellular spheroids was （42.9±4.0）% （P0.05）.Conclusion Multicellular spheroids of HT29 cells are an eligible model to mimic microtumors and avascular tumor regions in vivo,and helpful to research the structure formation and cell function crypt.

关 键 词：结肠癌 多细胞球 细胞黏附 耐药 

分 类 号：R73-35[医药卫生—肿瘤] R730.23[医药卫生—临床医学]