p53_k_ras基因型小鼠肺癌细胞株的建立与鉴定

《中国癌症杂志》2009年第19卷第5期

CHINA ONCOLOGY 2009 Vol.19 No.5

335p53-/-

,k-ras基因型小鼠肺癌细胞株的

建立与鉴定

［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-Loxp k-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。 ［关键词］　肺癌；　小鼠；　细胞株；　原代培养

中图分类号：R73-35+4 文献标识码：A 文章编号：1007-3639(2009)05-0335-05

Establishment of a mouse lung cancer cell line with the genotype of p53-/-,k-ras SUN Yi-hua,LI Li,PAN-Jun,YE Xiao-lei,WANG Zuo-yun,FENG Yan,CHEN Hai-quan (Department of Cardiothoracic Surgery, The Affiliated Sixth People's Hospital,Shanghai Jiao Tong University,Shanghai 200233,China)

Correspondence to：CHEN Hai-quan E-mail：hqchen1@

［Abstract ］ Background and purpose ：The lung cancer cell line with speci fi

c gene mutation is very important in research of lung cancer. k-ras an

d p53 play important roles in tumorgenesis, but ther

e was little research about mouse p53-/-,k-ras lung cancer cell line in China. We established mice lung cancer cell lines with the genotype o

f p53-/-,k-ras. Methods ：We induced lun

g cancer in p53loxp/loxp , Loxp-Stop-Loxp k-ras G12D transgenic mice by intranasal infection wit

h recombinant Cre adenoviruses that conditionally knock out p53 and activate k-ras in lung epithelium cells. Pathological histology of the mice lung cancer was ident

i fi ed by HE staining and con fi rmed 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. ［Key words ］　lung cancer; mouse; cell line; primary culture

通讯作者：陈海泉　E-mail:hqchen1@ 据世界卫生组织统计，肺癌已成为癌症中

的第一杀手，全球每年约新增1 200万的肺癌患者［1］，尽管有手术、放疗和化疗等综合治疗，但疗效仍然不佳，有报道显示1996年到2003年的肺癌5年生存率仅为16%［2］。进一步加强对

肺癌发生发展的机制研究，提出合理有效的治疗方案显得尤为重要。近年来，随着对肺癌研究的深入，国际上越来越多的使用转基因小鼠肺癌模型和特定基因变化的小鼠肺癌细胞株进行研究。k-ras激活和p53突变在肺癌细胞中较为常见，且p53-/-,k-ras的转基因小鼠模型研究显示这2个基因在肺癌的发生发展中均起着重要作