p53^(-/-),k-ras基因型小鼠肺癌细胞株的建立与鉴定  

Establishment of a mouse lung cancer cell line with the genotype of p53^(-/-),k-ras

作　　者：孙艺华[1,2] 李立[3] 潘俊[1] 叶小磊[3] 王作云[3] 冯艳[3] 陈海泉[1,2] 

机构地区：[1]上海交通大学附属第六人民医院胸心外科,上海200233 [2]复旦大学附属肿瘤医院胸外科,上海200032 [3]中国科学院上海生命科学研究院生物化学与细胞生物学研究所,上海200031

出　　处：《中国癌症杂志》2009年第5期335-339,共5页China Oncology

摘　　要：背景与目的:在肺癌的基础研究中,有特定基因变化的肺癌细胞株非常重要。k-ras激活和p53突变在肺癌的发生发展中有重要作用。本研究通过诱导小鼠肺癌生成,对肿瘤进行培养,建立p53-/-,k-ras基因型的小鼠肺癌细胞株。方法:用病毒吸入法,对p53loxp/loxp,Loxp-Stop-Loxpk-ras G12D转基因小鼠肺部细胞进行条件性基因敲除,诱导小鼠产生肺癌。通过HE染色和PCR法对小鼠肺癌组织进行组织病理学和基因型鉴定,对肿瘤细胞原代培养建立p53-/-,k-ras基因型的小鼠肺癌细胞株,并通过软琼脂克隆形成实验、生长曲线分析和核型分析对细胞株进行验证。结果:成功建立p53-/-,k-ras基因型的小鼠肺腺癌细胞株。p53-/-,k-ras基因型的小鼠肺癌细胞株体外生长速度与人肺腺癌细胞A549生长速度接近,能够在软琼脂中形成肉眼可见的克隆。结论:成功建立p53-/-,k-ras基因型的小鼠肺癌细胞株,为下一步的实验研究打下了基础。Background and purpose： The lung cancer cell line with specific gene mutation is very important in research of lung cancer, k-ras and p53 play important roles in tumorgenesis, but there was little research about mouse p53^-/-k-ras lung cancer cell line in China. We established mice lung cancer cell lines with the genotype of p53^-/-,k-ras. Methods： We induced lung cancer in p53^loxp/loxp, Loxp-Stop-Loxp k-ras^G12D transgenic mice by intranasal infection with recombinant Cre adenoviruses that conditionally knock out p53 and activate k-ras in lung epithelium cells. Pathological histology of the mice lung cancer was identified by HE staining and confirmed the genotype of the mouse lung cancer by PCR. A mouse lung cancer cell line was established by tissue primary cell culture. The anchorage-independent growth ability and proliferation rate were indentified compared with A549 cell line. Results： A mouse lung adenocarcinoma cell line with the genotype of p53^-/-, k-ras was established. This cell line had a similar proliferation rate with A549, and showed anchorage-independent growth in soft agar. Conclusion： We have successfully established a mouse lung cancer cell line with the genotype of p53^-/-, k-ras, which provided us a platform for further studying.

关 键 词：肺癌 小鼠 细胞株 原代培养 

分 类 号：R73-354[医药卫生—肿瘤]