CEA阳性细胞系的鉴定及其荷瘤小鼠模型建立

Identification of CEA-positive cell lines and establishment of their tumor-bearing mouse models

目的鉴定HT-29、MKN-45及Hela细胞系中CEA的阳性表达率,并分别建立3个细胞系对应的荷瘤小鼠肿瘤模型,分析其成瘤特性。方法采用RT-PCR、western blot检测HT-29、MKN-45及Hela细胞中的CEA表达情况,采用免疫荧光实验检测3株细胞系的CEA蛋白表达情况。以2×10^(7)/ml细胞浓度接种小鼠建立动物模型,观察荷瘤小鼠生长情况并记录肿瘤大小,对其成瘤特性进行分析。结果CEA蛋白在HT-29、MKN-45细胞中表达,而在Hela细胞中不表达,CEA蛋白主要存在于细胞的胞质中。MKN-45荷瘤小鼠的成瘤潜伏期为(6.3±1.5)d,HT-29成瘤潜伏期为(6.0±2.2)d,而Hela细胞的成瘤潜伏期为(10.0±1.6)d。MKN-45、HT-29细胞的成瘤时间明显短于Hela细胞。结论CEA在HT-29、MKN-45细胞中表达,而在Hela细胞中不表达。成功建立了3株细胞对应的荷瘤小鼠模型,为CEA阳性肿瘤的发病基础以及治疗提供了动物模型基础。

Objective To identify the expression of carcinoembryonic antigen(CEA)in the three cell lines of HT-29,MKN-45 and Hela,and establish tumor models of tumor-bearing mice corresponding to each cell line respectively,and then analyzed their tumorigenic characteristics.Methods RT-PCR and Western Blot were used to verify the CEA expression in HT-29,MKN-45 and Hela cells.The CEA protein of the three cell lines was verified and localized by immunofluorescence experiments.Finally,tumor-bearing mice were inoculated at a cell concentration of 2×10^(7)/ml.The growth of the tumor-bearing mice was observed and the tumor size was recorded,and their tumor-forming characteristics were analyzed.Results CEA protein was expressed in HT-29 and MKN-45 cells,but not in Hela cells.CEA protein mainly existed in the cytoplasm of cells.All tumor-bearing mice in the experimental group survived and formed tumors.The tumorigenesis latency of MKN-45 tumor-bearing mice was(6.3±1.5)d,the tumorigenesis latency of HT-29 was(6.0±2.2)d,and the tumorigenesis of Hela cells The incubation period was(10.0±1.6)d.Conclusions CEA is expressed in HT-29 and MKN-45 cells,but not in Hela cells.The tumor-bearing mouse models corresponding to the three cell lines have been successfully established.It provides an animal model basis for the pathogenesis and treatment of CEA-positive tumors.