食管癌te-1细胞生物学特性相关性研究-correlation study on biological characteristics of esophageal cancer te - 1 cells.docxVIP

I I 摘 要 目的：研究食管癌 TE-1 细胞的生物学性状。 方法：（1)将购自上海生命科学院的人食管 TE-1 细胞株进行 HE 染色，（2)用 MTT 法绘 制细胞生长曲线；(3)免疫组织化学检测细胞特异性免疫标志；（4）流式细胞仪分析细 胞周期（5）采用 MTT 法观察顺铂，紫杉醇，甲氨蝶呤以及氟尿嘧啶对细胞的毒性作用， 同时计算半数抑制率；（6）用紫杉醇的（0.8μg/ml）半数致死量浓度作用细胞，观察 细胞是否产生耐药性。 结果：（1）用 DMEM 高糖培养基培养，生长良好。HE 染色下，细胞大致分为三种：一种 细胞呈椭圆形，核大，胞浆少，一种细胞呈三角形或多角形，大小，形态不等，一种细 胞有双核或者多核，核大深染，染色质丰富，核分裂相多见；（2）细胞接种培养后第 1,2 天为生长潜伏期，第 3-6 天为指数增生期，第 7 天后进入平台期，其生长曲线呈近 似 S 形。免疫组织化学检测 3 种抗体均表达阳性，其中 CK7 阳性表达定位于胞浆，Ki-67 定位于胞核，p53 定位于胞核；（3）TE-1 细胞周期：半数细胞处于增生期，半数处于 静止期；（4）四种化疗药物毒性作用方式不同，其中紫杉醇对 TE-1 细胞的毒性作用具 有明显的浓度和时间依赖性；(5)食管 TE-1 细胞在紫杉醇（0.8μg/ml）作用下，168 小时细胞全部死亡，产生耐药性。 结论：（1） TE-1 细胞符合食管鳞癌的生物学性状；（2）不同的化疗药物对食管癌 TE-1 细胞作用方式不同，紫杉醇对 TE-1 细胞的作用具有明显的浓度和时间依赖性且易产生 耐药性。 关键词：食管癌 TE-1 细胞株 生物学特性 化疗药物 紫杉醇 论文类型：A（基础研究） II II Abstract Objective: Study the biological characteristics of esophageal cancer cells Methods: (1) The morphological characteristicas was observed in HE staining.（2）The cell grouth curve was analysed by MTT method.（3）The immunocytochemical method was employed to detect the specific cell phenotype （; 4）The cell cycle was analysed by FCM（. 5） The toxicity of cisplatin, paclitaxel, methotrexate and fluorouracil to TE-1 cells were analysed by MTT and the LC50 of paclitaxelwas calculated. Results: The cells grew well in the high glucose DMEM,. The cells can be divided into three of kinds in HE staining，the first cells were oval-shaped, nucleus large and small cytoplasm, the second were triangular or polygonal, and the third have dual-nucleus or multi -nucleus and nucleus mitotic figures were common. After the cells were planted, the cells in the first-second day were in the incubation period, the third-fifth day was in proliferative index period and in the seventh day, the cells were into the plateau growth period. The CK7 expression was positive in the cytoplasm. Ki-67 positive expression were in the nucleus. P53 positive expression were in the nucleus too. Half of the cells were in the proliferative phase of cell cycle and half in the quiescent p