HER2高表达的人乳腺癌原代细胞模型的建立  

Establishment of primary Her-2 overexpression human breast cancer cell model

作　　者：王丽娟[1] 赵悦[1] 肖华卫[1] 白娥[1] 董超[2] 杨韬[3] 杨安钢[3] 朱青[1] 

机构地区：[1]西安交通大学医学院第一附属医院肿瘤中心肿瘤内科,陕西西安710061 [2]第四军医大学西京脑科医院神经外科,陕西西安710032 [3]第四军医大学免疫学教研室,陕西西安710032

出　　处：《细胞与分子免疫学杂志》2011年第10期1148-1150,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(810721117)

摘　　要：目的:探讨人乳腺癌恶性胸水肿瘤细胞的分离、纯化、培养与肿瘤细胞的鉴定,为HER2阳性的人乳腺癌的研究提供肿瘤原代细胞模型。方法:利用淋巴细胞分离液及密度梯度离心法分离、纯化恶性胸水肿瘤细胞,分离得到的细胞培养并传至第5代,通过流式细胞仪分选肿瘤细胞、Q-PCR技术、Western blot技术验证细胞HER2分子的表达水平、通过裸鼠荷瘤实验检测细胞的成瘤能力。结果:淋巴细胞分离液及密度梯度离心法分离得到乳腺癌原代细胞;流式细胞分选结果显示未分选出HER2高表达的肿瘤细胞;Q-PCR及Western blot检测发现分离得到的细胞在mRNA及蛋白水平HER2的表达水平高于乳腺癌MCF-7细胞;裸鼠荷瘤实验结果显示分离得到的细胞可以成功荷瘤。结论:胸水中分离得到的细胞为HER2阳性的乳腺癌原代细胞;对于原代细胞的鉴定需要采用不同的方法加以论证。AIM： Through isolation and purification primary HER2 overexpression human breast cancer cells from malignant pleural effusion and identification the HER2 expression level of the cells to establish the primary HER2 overexpression human breast cancer cell model. METHODS： Malignant pleural effusion of HER2 overexpression breast cancer patient was collected. The primary cells were extracted from malignant pleural effusion by Lymphocyte separation medium and the method of density gradient centrifugation. When the primary cells were cultured and spreaded to the 5th generation, the HER2 expression level of the primary cells were detected by the methods of Q-PCR, Western blot and flow cytometry （FCM）. Ability of tumor-bearing was detected by tumor-bearing nude mice assay. RESULTS： The primary HER2 overexpression human breast cancer cells were extracted and identified by the methods of Q-PCR, Western blot and tumor-bearing nude mice assay, even though the FCM showed Negative results. CONCLUSION： The primary HER2 overexpression human breast cancer cell model was established; Identification of primary cells need to be confirmed by different methods.

关 键 词：恶性胸水 HER2阳性的乳腺癌细胞 密度梯度离心法 裸鼠荷瘤 

分 类 号：R730.43[医药卫生—肿瘤]